NATIONAL TECHNICAL REGULATION
QCVN 4-10:2010/BYT
ON FOOD ADDITIVES - COLOURS
Foreword
QCVN 4-10:2010/BYT is developed by
the Drafting Board for National technical regulation on food additives and food
processing aids, submitted by the Vietnam Food Administration for approval and
promulgated together with the Circular No. 27/2010/TT-BYT dated May 20, 2010 of
the Minister of Health.
NATIONAL
TECHNICAL REGULATION ON FOOD ADDITIVES - COLOURS
I. GENERAL
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This National technical regulation
(hereinafter referred to as “the Regulation”) provides for specifications and
regulatory requirements for quality, hygiene and safety of colours used as food
additives.
2. Regulated entities
This Regulation applies to:
2.1. Importers, exporters,
producers, traders and users of colours used as food additives (hereinafter
referred to as “entities”).
2.2. Relevant regulatory bodies.
3. Interpretation of terms and
acronyms:
3.1. “colour” means a food additive
used for the purpose of adding or restoring color in a food product.
3.2. JECFA monograph 1 - Vol. 4
(JECFA monographs 1 - Combined compendium of food additive specifications;
Joint FAO/WHO expert committee on food additives; Volume 4 - Analytical
methods, test procedures and laboratory solutions used by and referenced in the
food additive specifications; FAO, 2006) is developed by JECFA and
was published by FAO in 2006.
3.3. “C.A.S number”
(Chemical Abstracts Service) refers to registry numbers of chemical substances
identified by the American Chemical Society.
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3.5. “ADI” stands
for Acceptable daily intake.
3.6. “INS” stands
for international numbering system.
II.
SPECIFICATIONS, TESTS AND SAMPLING
1. Specifications
and tests for flavour enhancers specified in the annexes to this Regulation:
1.1. Annex 1: Specifications and
tests for curcumin
1.2. Annex 2: Specifications and
tests for riboflavin
1.3. Annex 3: Specifications and
tests for tatrazine
1.4. Annex 4: Specifications and
tests for quinoline
1.5. Annex 5: Specifications and
tests for sunset yellow FCF
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1.7. Annex 7: Specifications and
tests for carmoisine
1.8. Annex 8: Specifications and
tests for amaranth
1.9. Annex 9: Specifications and
tests for ponceau 4R
1.10. Annex 10: Specifications and
tests for erythrosine
1.11. Annex 11: Specifications and
tests for red 2G
1.12. Annex 12: Specifications and
tests for allura red AC
1.13. Annex 13: Specifications and
tests for indigotine
1.14. Annex 14: Specifications and
tests for brilliant blue FCF
1.15. Annex 11: Specifications and
tests for chlorophylls
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1.17. Annex 17: Specifications and
tests for chlorophyllins, copper complexes sodium and potassium salts
1.18. Annex 18: Specifications and
tests for green S
1.19. Annex 19: Specifications and
tests for caramel
1.20. Annex 20: Specifications and
tests for brilliant black PN
1.21. Annex 21: Specifications and
tests for brown HT
1.22. Annex 22: Specifications and tests
for synthetic β-carotene
1.23. Annex 23: Specifications and
tests for carotenes, vegetable
1.24. Annex 24: Specifications and
tests for annatto extracts
1.25. Annex 25: Specifications and
tests for β-Apo-carotenal
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1.27. Annex 27: Specifications and
tests for grape skin extract
1.28. Annex 28: Specifications and
tests for titanium dioxide
1.29. Annex 29: Specifications and
tests for iron oxides
1.30. Annex 30: Specifications and
tests for fast green FCF
2. Specifications
specified in this Regulation are tested under JECFA monograph 1 - Vol. 4,
except for some specific tests described in the annexes. The tests provided in
this Regulation are optional. Other equivalent tests may be used.
3. Sampling
adheres to the guidelines in the Circular No. 16/2009/TT-BKHCN dated June 02,
2009 by the Ministry of Science and Technology and relevant regulations of law.
III. REGULATORY REQUIREMENTS
1. Declaration of conformity
1.1. Conformity of colours shall be
declared in accordance with the regulations set out in this Regulation.
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2. Inspection of colours
The quality, hygiene and safety of
colours must be inspected in accordance with the regulations of law.
IV. RESPONSIBILITIES OF ENTITIES
1. Entities must declare conformity according to the specifications
mentioned in this Regulation, register their declarations of conformity at the
Vietnam Food Administration and ensure the quality, hygiene and safety as
declared.
2. Entities are entitled to import, export, produce, sell and use colours
only after their completion of registration of declarations of conformity and
their compliance with regulations of law on quality, hygiene, safety and
labeling.
V. IMPLEMENTATION
1. The
Vietnam Food Administration shall preside over and cooperate with competent
authorities concerned to provide guidance on and organize the implementation of
this Regulation.
2. The Vietnam
Food Administration shall, according to its managerial duties, suggest
amendments to this Regulation to the Ministry of Health.
3. In the
cases where any of the international guidelines for tests and regulations of
law referred to in this Regulation is amended or replaced, the newest one shall
apply.
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ANNEX 1
SPECIFICATIONS AND TESTS FOR CURCUMIN
1. Synonyms
Turmeric yellow, Kurkum
INS: 100i
ADI = 0 – 3mg/kg bw.
2. Definition
Curcumin is obtained by solvent
extraction of turmeric i.e., the ground rhizomes of Curcuma longa L. (Curcuma
domestica Valeton). In order to obtain a concentrated curcumin powder,
the extract is purified by crystallization. The product consists essentially
of curcumins; i.e. the colouring principle
1,7-bis-(4-hydroxy-3-methoxy-phenyl)-hepta-1,6-diene3,5-dione (synonyms:
Curcumin, Diferuloylmethane, CI Natural Yellow 3, CI (1975) 75300) and its
desmethoxy- and bis-desmethoxy-derivatives in varying proportions. Minor
amounts of oils and resins naturally occurring in turmeric may be present. Only
the following solvents may be used in the extraction and purification:
acetone, methanol, ethanol, isopropanol, hexane and ethyl acetate. Supercritical
carbon dioxide may also be used in the extraction.
Chemical names
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I.
1,7-Bis-(4-hydroxy-3-methoxyphenyl)-hepta-1,6-diene-3,5-dione
II.
1-(4-Hydroxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)-hepta-1,6-diene3,5-dione
III.
1,7-Bis-(4-hydroxyphenyl)-hepta-1,6-diene-3,5-dione
C.A.S.
number
I.
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II.
33171-16-3
III.
33171-05-0
Chemical
formula
I.
C21H20O6
II.
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III.
C19H16O4
Structural
formula
I.
R1= R2=OCH3
II.
R1= OCH3;
R2=H
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R1= R2=H
Formula
weight
I.
368.39
II.
338.39
III.
308.39
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3. Description
Orange-yellow crystalline powder.
4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Insoluble in water and in diethyl
ether; soluble in ethanol and in glacial acetic acid.
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A solution of the sample in
ethanol is characterized by a pure yellow colour and a light green
fluorescence. Addition of this ethanol extract to concentrated sulfuric acid
produces a deep crimson colour.
Thin
Layer Chromatography
Passes test (described in
“Tests”).
5.2.
Purity
Residual
solvents
Acetone:
Not more than 30 mg/kg
Hexane:
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Methanol:
Ethanol:
Isopropanol:
Ethyl
acetate:
Lead
Not more than 2 mg/kg
5.3.
Content
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6. Tests
6.1.
Identification
Colour
reactions
Treat an aqueous or dilute
ethanolic solution of the sample with hydrochloric acid until a slightly
orange colour begins to appear. Divide the mixture into 2 parts and add some
boric acid powder or crystals to one portion. A marked reddening will be
quickly apparent, best seen by comparison with the portion to which the boric
acid has not been added. The test may also be made by dipping pieces of
filter paper in an ethanolic solution of colouring matter, drying at 100º,
and then moistening with a weak solution of boric acid to which a few drops
of hydrochloric acid have been added. On drying, a cherry-red colour will
develop.
Thin
Layer Chromatography
Spot 5 µl of test solution (0.01
g of sample in 1 ml of 95% ethanol) on a TLC (Microcrystalline cellulose, 0.1
mm) plate. Develop the plate in a chamber containing a mixture of
3-methyl-1-butanol/ethanol/ water/ammonia (4:4:2:1) as solvent and allow the
solvent front to ascend 10-15 cm. Examine under daylight and under UV-light
and observe:
Two or three yellow spots with Rf
between 0.2 and 0.4 under daylight and UV-light.
Spots with Rf about
0.6 and 0.8 under UV-light
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6.2
Purity
Lead
Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
6.3.
Method of assay
Accurately weigh about 0.08 g of
the sample in a 200-ml volumetric flask and dissolve by shaking with ethanol.
Make up to volume with ethanol and mix. Pipette 1.0 ml of solution into a
100-ml volumetric flask and make up to volume with ethanol.
Determine the absorbance (A) at
425 nm in a 1-cm cell. Calculate the total colouring matters content of the
sample using the following equation:
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Where:
A = absorbance of sample
W = weight of sample
1607 = specific absorbance of the
curcumin standard in ethanol at 425 nm
The determination must be
performed without delay, because the colour fades.
ANNEX 2
SPECIFICATIONS AND TESTS FOR RIBOFLAVIN
1. Synonyms
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INS: 101i
ADI = 0 –
0.5 mg/kg bw.
2. Definition
Chemical
names
Riboflavin;
3,10-dihydro-7,8-dimethyl-10-[(2S,3S,4R)-2,3,4,5-
tetrahydroxypentyl]benzo-[g]pteridine-2,4-dione; 7,8-dimethyl-10-(1'-Dribityl)isoalloxazine.
C.A.S.
number
83-88-5
Chemical
formula
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Structural
formula
Formula
weight
376.37
3. Description
Yellow to
orange-yellow crystalline powder, with slight odour.
4. Functional
uses
Colour.
5. Specifications
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Solubility
Very slightly soluble in water;
practically insoluble in alcohol, chloroform, acetone and ether; very soluble
in dilute alkali solutions.
Spectrophotometry
Using the aqueous solution from
the Assay, determine the absorbance (A) at 267 nm, 375 nm and 444 nm.
The ratio A375/A267
is between 0.31 and 0.33.
The ratio
A444/A267 is between 0.36 and 0.39.
Specific
rotation
[a]20, D: Between -115o
and -140o.
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Passes test (described in
“Tests”).
5.2. Purity
Loss on
drying
Not more than 1.5%.
Sulfated
ash
Not more than 0.1%.
Subsidiary
colouring matters
Passes test (described in
“Tests”).
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Not more than 100 mg/kg
calculated as aniline.
Lead
Not more than 2 mg/kg.
5.3. C17H20N4O6
content
Not less than 98%.
6. Tests
6.1.
Identification
Specific
rotation
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Colour
reaction
Dissolve about 1 mg of sample in
100 ml of water. The solution has a pale greenish-yellow colour by
transmitted light, and by reflected light has an intense yellowish-green
fluorescence which disappears on the addition of mineral acids and alkalis.
6.2.
Purity
Loss on
drying
At 105o, 4 h
Sulfated
ash
- Tested as directed under JECFA
monograph 1 - Vol. 4.
- Test 2 g of the sample
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Prepare the standard for this
test for the absence of lumiflavin by diluting 3 ml of 0.1 N potassium
dichromate with water to 1000 ml. Pour some chloroform through an alumina
column to remove any ethanol. To 10 ml of this chloroform add 35 mg of the
sample, shake for 5 min and filter. The colour of the filtrate should be no
more intense than that of 10 ml of the standard when viewed in identical
containers.
Lead
Tested as directed under JECFA
monograph 1 - Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in Annex 1: General tests, “Instrumental Methods.”
6.3.
Method of assay
Carry out the assay in subdued
light. In a brown glass 500 ml volumetric flask, suspend 65.0 mg of the
sample in 5 ml of water, ensuring that it is completely wetted, and dissolve
in 5 ml of 2 N sodium hydroxide solution. As soon as dissolution is complete,
add 100 ml of water and 2.5 ml of glacial acetic acid and dilute to 500.0 ml
with water. Place 20.0 ml of this solution in a brown glass 200 ml volumetric
flask, add 3.5 ml of a 1.4% w/v solution of sodium acetate and dilute to
200.0 ml with water. Measure the absorbance (A) at the maximum at 444 nm.
%
Riboflavin = [(A×5000) / (328×W)] × 1,367
Where:
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W = weight of sample in g.
ANNEX 3
SPECIFICATIONS AND TESTS FOR TARTRAZINE
1. Synonyms
CI Food
Yellow 4, FD&C Yellow No. 5; CI (1975) No. 19140
INS: 102
ADI = 0 -
7.5 mg/kg bw.
2. Definition
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May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
Trisodium4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-
1H-pyrazole-3-carboxylate
C.A.S.
number
1934-21-0
Chemical
formula
C16H9N4Na3O9S2
Structural
formula
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534.37
3. Description
Light
orange powder or granules.
4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
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Identification
of colouring matters
Passes
test.
5.2. Purity
Loss on
drying at 135 °C
Not more
than 15% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
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Subsidiary
colouring matters
Not more
than 1%.
Organic
compounds other than colouring matters
Not more
than 0.5% sum of 4-hydrazinobenzenesulfonic acid, 4- aminobenzenesulfonic
acid, 5-oxo-1-(4-sulfophenyl)-2-pyrazoline-3- carboxylic acid,
4,4'-(diazoamino)dibenzenesulfonic acid, tetrahydroxysuccinic acid.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
Ether
extractable matter
Not more
than 0.2%.
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Not less
than 85% total colouring matters
6. Tests
6.1.
Purity
Lead
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Determine
using an atomic absorption technique appropriate to the specified level. The
selection of sample size and method of sample preparation may be based on the
principles of the method described in JECFA monograph, Volume 4,
“Instrumental Methods.”
Subsidiary
colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
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Organic
compounds other than colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Use
HPLC under the following conditions: HPLC elution gradient: 2 to 100% at 2%
per min.
6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (in Annex 1: General tests),
using the following:
Weight of
sample: 0.6 - 0.7 g;
Buffer:
15 g sodium hydrogen tartrate;
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ANNEX 4
SPECIFICATIONS AND TESTS FOR QUINOLINE
1. Synonyms
CI Food Yellow 13, CI (1975) No.
47005,
INS: 104
ADI = 0 – 10 mg/kg bw.
2. Definition
Prepared by sulfonating
2-(2-quinolyl)-1,3-indandione or a mixture containing about two-thirds
2-(2-quinolyl)-1,3-indandione and one-third 2-
[2-(6-methyl-quinolyl)]1,3-indandione; consists essentially of sodium salts
of a mixture of disulfonates (principally), monosulfonates and trisulfonates
of the above compounds and subsidiary colouring matters together with sodium
chloride and/or sodium sulfate as the principal uncoloured components.
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Chemical names
Disodium
2-(1,3-dioxo-2-indanyl)-6,8-quinolinesulfates; disodium 2-(2-
quinolyl)-indan-1,3-dionedisulfonates (principal component)
C.A.S.
number
8004-72-0 (Unmethylated
disulfonic acids)
Chemical
formula
C18H9NNa2O8S2
(principal component)
Structural
formula
Principal component
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8 salt: R1, R2 =
SO3H
Formula
weight
477.38 (Principal components)
3. Description
Yellow powder or granules.
4. Functional
uses
Colour.
5. Specifications
5.1.
Identification
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Solubility
Soluble in water; sparingly
soluble in ethanol
Identification
of colouring matters
Passes test.
5.2.
Purity
Loss on
drying at 135°C
Not more than 30% together with
chloride and sulfate calculated as sodium salts.
Water
insoluble matter
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Lead
Not more than 2 mg/kg.
Zinc
Not more than 50 mg/kg.
Subsidiary
colouring matters
Not more than 4 mg/kg: of
2-(2-quinolyl)-1,3-indandione and 2-[2-(6- methylquinolyl)]-1,3-indandione
Organic
compounds other than colouring matters
Not more than 0.5%, sum of
2-methylquinoline, 2-methylquinolinesulfonic acid, phthalic acid,
2,6-dimethylquinoline, 2,6-dimethylquinolinesulfonic acid
Unsulfonated
primary aromatic amines
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Ether
extractable matter
Not more than 0.2%.
5.3.
Content
Not less than 70% total colouring
matters. Quinoline Yellow prepared from 2-(2-quinolyl)-1,3-indandione (only)
shall have the following composition:
- not less than 80% shall be
disodium 2-(2-quinolyl)-indan1,3-dionedisulfonates;
- not more than 15% shall be
sodium 2-(2-quinolyl)-indan-1,3- dionemonosulfonates;
- not more than 7% shall be
trisodium 2-(2-quinolyl)-indan-1,3- dionetrisulfonate;
6. Tests
6.1.
Purity
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Lead
Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph, Volume 4, “Instrumental Methods.”
Subsidiary
colouring matters
Limit test for
2-(2-quinolyl)-1,3-indandione and 2-[2-(6-methyl-quinolyl)]- 1,3-indandione.
Use the apparatus and ether
quality described in Ether Extractable Matter and carry out an extraction
following the details given there under. Wash the ether extract with two
25-ml portions of water. Evaporate the ether extract to about 5 ml and then
transfer it to an oven at 105o to remove the remaining ether. Dissolve
the residue in chloroform, and dilute the solution to exactly 10 ml. Determine
the absorbance at the wavelength of maximum absorption (approximately 420 nm)
using chloroform as the reference solution. The absorbance corresponding to
the limit figure of 4 mg/kg 2-(2-quinolyl-1,3-indandione is 0.27. Any
2-[2-(6-methylquinolyl)]- 1,3-indandione is assessed as
2-(2-quinolyl)-1,3-indandione.
Organic
compounds other than colouring matters
Organic compounds other than
colouring matters in Quinoline prepared from 2-(2-quinolyl)-1,3-indandione
(only).
Determine by liquid chromatography,
using the following conditions:
Instrument: High performance
liquid chromatograph fitted with a gradient elution accessory.
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Column: 250 x 4 nm, Nucleosil
C18, 7 µm.
Solvent system:
A: Acetate Buffer pH 4.6: water
(1:10) (Acetate Buffer is 1M NaOH:1 M acetic acid:water (5:10:35).
B: (A):methanol (20:80).
Sample concentration: 1%
weight/volume in Solvent A.
Gradient:
Min
%A
%B
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100
0
15
65
35
20
50
50
25
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100
36
0
100
42
100
0
Flow rate: 1 ml/min
6.2.
Method of assay
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Proceed as directed under Total
Content by Spectrophotometry (JECFA monograph 1 - Vol. 4).
Solvent: pH 7 phosphate buffer
Dilution of solution A: 10 ml to
250 ml
Absorptivity (a): 86.35
Approximate wavelength of maximum
absorption: 415 nm
Determination of the percentages
of di-, mono- and trisulfonates in Quinoline prepared from
2-(2-quinolyl)-1,3-indandione (only): Use the HPLC conditions prescribed in
the Determination of Organic Compounds other than Colouring Matters with a
sample solution of concentration 0.05% in HPLC Solvent A in place of the
sample solution of concentration 1%. Express the results as percentages of
the Total colouring matters present.
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SPECIFICATIONS AND TESTS FOR SUNSET YELLOW FCF
1. Synonyms
CI Food
Yellow 3, FD&C Yellow No. 6; CI (1975) No. 15985
INS: 110
ADI = 0 -
2.5 mg/kg bw.
2. Definition
Consists essentially of disodium
6-hydroxy-5-(4-sulfonatophenylazo)-2- naphthalene-6-sulfonate and subsidiary
colouring matters together with NaCl and/or Na2SO4 as
the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
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C.A.S.
number
2783-94-0
Chemical
formula
C16H10N2Na2O7S2
Structural
formula
Formula
weight
452.38
3. Description
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4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Soluble
in water; sparingly soluble in ethanol.
Identification
of colouring matters
Passes
test.
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Loss on
drying at 135°C
Not more
than 15% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
Not more
than 5%.
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Organic
compounds other than colouring matters
Not more
than 0.5%, sum of 4-amino-1-benzenesulfonic acid;
3-hydroxy2,7-naphthalenedisulfonic
acid;
6-hydroxy-2-naphthalenesulfonic
acid;
7-
hydroxy-1,3-naphthalenedisulfonic acid;
4,4'-
diazoaminodibenzenesulfonic acid;
6,6'-oxydi-2-naphthalenesulfonic
acid.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
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Not more
than 0.2%.
5.3. Content
Not less
than 85% total colouring matters
6. Tests
6.1.
Purity
Lead
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Determine
using an atomic absorption technique appropriate to the specified level. The
selection of sample size and method of sample preparation may be based on the
principles of the method described in JECFA monograph 1-Vol. 4, “Instrumental
Methods.”
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- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 4; Height of ascent of solvent front: approximately
17 cm.
Organic
compounds other than colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Proceed as directed under
Determination by High Performance Liquid Chromatography using an elution
gradient of 2 to 100% at 4% per min followed by elution at 100%.
6.2.
Method of assay
Proceed
as directed under Total Content by Titration with Titanous Chloride (under
JECFA monograph 1-Vol. 4), using the following:
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Buffer:
10 g sodium citrate;
Weight
(D) of colouring matters equivalent to 1.00 ml of 0.1 N TiCl3:
11.31 mg.
ANNEX 6
SPECIFICATIONS AND TESTS FOR CARMINES
1. Synonyms
Cochineal carmin, CI Natural Red
4, CI (1975) No. 75470,
INS 120
A group ADI of 0-5 mg/kg bw for
carmines, as ammonium carmine or the equivalent of Ca, K and Na salts.
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Obtained by aqueous extraction of
cochineal, which consists of the dried bodies of the female insect Dactylopius
coccus Costa; the colouring principle is a hydrated aluminium chelate of
carminic acid in which aluminium and carminic acid are thought to be present
in the molar ratio 1:2.
In commercial products the
colouring principle is present in association with ammonium, calcium,
potassium or sodium cations, singly or in combination, and these cations may
also be present in excess. Products may also contain proteinaceous material
derived from the source insect, and may also contain free carminate or a
small excess of aluminium cations.
Chemical
names
Hydrated aluminium chelate of
carminic acid (7-beta-D-glucopyranosyl3,5,6,8-tetrahydroxy-1-methyl-9,10-dioxo-anthracene-2-carboxylic
acid)
C.A.S.
number
1390-65-4 (carmine)
1260-17-9 (carminic acid)
Chemical
formula
Carminic acid: C22H20O13
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Carminic acid
Aluminium complex of carminic
acid
Formula
weight
Carminic acid: 492.39
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Red to dark red, crumbly solid or
powder
4. Functional uses
Colour
5. Specifications
5.1.
Identification
Solubility
The solubility of carmine
preparations varies depending on the nature of the cations present. Products
where the major cation is ammonium (ammonium carminate) are freely soluble in
water at pH 3.0 and pH 8.5. Products where the major cation is calcium
(calcium carminate) are very slightly soluble in water at pH 3.0 but freely
soluble at pH 8.5.
Colour
reaction
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5.2.
Purity
Loss on
drying
Not more than 20%.
Total ash
Not more than 12%.
Protein
Not more than 25%.
Matter insoluble in dilute
ammonia
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Lead
Not more than 2 mg/kg.
Microbiological criteria
Salmonella: Negative per
test.
5.3. C22H20O13
content
Not less than 50% of C22H20O13
on the dry basis.
6. Tests
6.1 Identification
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Colour
reactions
- Make a solution of the sample
slightly alkaline by adding 1 drop of 10% sodium hydroxide or potassium
hydroxide solution. A violet colour is produced.
- Add a small sodium dithionite
(Na2S2O4) crystal to acid, neutral or
alkaline solutions of the sample. The solutions are not decolourized.
- Dry a small quantity of the
sample in a porcelain dish. Cool thoroughly and treat the dry residue with 1
or 2 drops of cold sulfuric acid TS. No colour change occurs.
- Acidify a dispersion of the
sample in water with 1/3 volume of hydrochloric acid TS and shake it with
amyl alcohol. Wash the amyl alcohol solution 2-4 times with an equal volume
of water to remove hydrochloric acid. Dilute the amyl alcohol solution with
1-2 volumes of petroleum ether (40-60o) and shake with a few small
portions of water to remove colour. Add, dropwise, 5% uranium acetate, shaking
thoroughly after each addition. A characteristic emerald-green colour is
produced.
6.2 Purity
Loss on
drying
- Tested as directed under JECFA
monograph 1-Vol. 4.
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Total ash
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Test 1 g of the sample as
directed in the test for Ash.
Protein
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Proceed as directed under
Nitrogen Determination (non-ammonia N x 6.25)
Matter insoluble in dilute
ammonia
Dissolve about 0.25 g of the
sample, previously dried and accurately weighed, in 2.5 ml of dilute ammonia
solution (160 ml of strong ammonia TS, made up to 500 ml) and dilute to 100
ml with water: the solution is clear. Filter through a sintered glass filter.
Wash with a 0.1% ammonia solution and dry to constant weight at 105º.
Lead
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6.3. Method of assay
Weigh accurately about 100 mg of
the sample, dissolve in 30 ml of boiling 2N hydrochloric acid and cool. Transfer
quantitatively to a 1000-ml volumetric flask, dilute to volume with water,
and mix. Determine the absorbance of the solution in a 1 cm cell at the
wavelength of maximum absorbance (about 494 nm) using water as the blank. Calculate
the percentage of carminic acid in the sample analysis using the formula:
(100 × A
× 100) / (1,39 × W)
where:
A = absorbance of the sample
solution;
W = weight, in mg, of the sample
taken; and
1.39 = absorbance of a solution
of carminic acid having a concentration of 100 mg per 1000 ml;
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ANNEX 7
SPECIFICATIONS AND TESTS FOR CARMOISINE
1. Synonyms
CI Food
Red 3, Azorubine; CI (1975): 14720
INS: 122
ADI = 0 -
4 mg/kg bw.
2. Definition
Consists essentially of disodium
4-hydroxy-3-(4-sulfonato-1-naphthylazo)- 1-naphthalenesulfonate and
subsidiary colouring matters together with NaCl and/or Na2SO4
as the principal uncoloured components.
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Chemical
names
Disodium
4-hydroxy-3-(4-sulfonato-1-naphthylazo)-1-naphthalenesulfonate.
C.A.S.
number
3567-69-9
Chemical
formula
C20H12N2Na2O7S2
Structural
formula
Formula
weight
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3. Description
Red
powder or granules.
4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Soluble
in water; sparingly soluble in ethanol.
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Passes
test.
5.2. Purity
Loss on
drying at 135 °C
Not more
than 15% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
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Not more than 1%.
Organic
compounds other than colouring matters
Not more than 0.5% of
4-Amino-1-naphthalenesulfonic acid and 4-Hydroxy-1-naphthalenesulfonic acid
together.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
Ether
extractable matter
Not more
than 0.2%.
5.3. Content
Not less
than 85% total colouring matters.
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6.1
Purity
Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 4; Height of ascent of solvent front: approximately
17 cm.
Organic
compounds other than colouring matters
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- Use HPLC under the following
conditions: HPLC elution gradient: 1 to 100% at 2.0% per min.
6.3.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 0.5 - 0.6 g;
Buffer:
15 g sodium hydrogen tartrate;
Weight (D) of colouring matters
equivalent to 1.00 ml of 0.1 N TiCl3: 12.56 mg
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SPECIFICATIONS AND TESTS FOR AMARANTH
1. Synonyms
CI Food
Red 9, Naphtol Rot S.; CI (1975): 16185
INS: 123
ADI = 0 -
0.5 mg/kg bw.
2. Definition
Consists essentially of trisodium
3-hydroxy-4-(4-sulfonato-1-naphthylazo)- 2,7-naphthalenedisulfonate and
subsidiary colouring matters together with sodium chloride and/or sodium
sulfate as the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
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C.A.S.
number
915-67-3
Chemical
formula
C20H11N2Na3O10S3
Structural
formula
Formula
weight
604.48
3. Description
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4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Soluble
in water; sparingly soluble in ethanol
Identification
of colouring matters
Passes
test.
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Loss on
drying at 135 °C
Not more
than 15% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
Not more
than 3%.
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Not more
than 0.5%, sum of 4-amino-1-benzenesulfonic acid;
3-
hydroxy-2,7-naphthalenedisulfonic acid, 6-hydroxy-2-naphthalenesulfonic acid,
7-hydroxy-1,3-naphthalenedisulfonic acid and 7-hydroxy-1,3,6-
naphthalene-trisulfonic acid.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
Ether
extractable matter
Not more
than 0.2%.
5.3. Content
Not less
than 85% total colouring matters.
6. Tests
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Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Use the
following conditions: Developing solvent: No. 4; Height of ascent of solvent
front: approximately 17 cm, then 1 h further development.
Organic
compounds other than colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
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6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 0.7 - 0.8 g;
Buffer:
10 g sodium citrate;
Weight (D) of colouring matters
equivalent to 1.00 ml of 0.1 N TiCl3: 15.11 mg.
ANNEX 9
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1. Synonyms
CI Food
Red 7, Cochineal Red A ; New Cochineal;
CI
(1975): 16255
INS: 124
ADI = 0 -
4 mg/kg bw.
2. Definition
Consists essentially of trisodium
d-2-hydroxy-1-(4-sulfonato-1- naphthylazo)-6,8-naphthalenedisulfonate, and
subsidiary colouring matters together with sodium chloride and/or sodium
sulfate as the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
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C.A.S.
number
2611-82-7
Chemical
formula
C20H11N2Na3O10S3.1,5H2O
Structural
formula
Formula
weight
631.51
3. Description
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4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Soluble
in water; sparingly soluble in ethanol.
Identification
of colouring matters
Passes
test.
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Loss on
drying at 135 °C
Not more
than 20% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
Not more
than 1%.
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Not more
than 0.5% of sum of: 4-amino-1-naphthalenesulfonic acid, 7-hydroxy-1,3-
naphthalenedisulfonic acid, 3-hydroxy-2,7-naphthalenesulfonic acid, 6-
hydroxy-2-naphthalenesulfonic acid, 7-hydroxy-1,3,6- naphthalenetrisulfonic
acid.
Unsulfonated primary
aromatic amines
Not more than 0.01% calculated as
aniline.
Ether
extractable matter
Not more than 0.2%.
5.3. Content
Not less
than 85% total colouring matters
6. Tests
6.1.
Purity
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Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Use the
following conditions: Developing solvent: No. 3; Height of ascent of solvent
front: approximately 17 cm, then 1 h further development
Organic
compounds other than colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Use
HPLC under the following conditions: HPLC elution gradient: 2 to 100% at 2%
per min.
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Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of sample: 0.7 - 0.8 g;
Buffer: 10 g sodium citrate;
Weight (D) of colouring matters
equivalent to 1.00 ml of 0.1 N TiCl3: 15.78 mg.
ANNEX 10
SPECIFICATIONS AND TESTS FOR ERYTHROSINE
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CI Food
Red 14, FD&C Red No. 3; CI (1975): 45430
INS 127
ADI = 0 -
0,1 mg/kg bw
2. Definition
Consists essentially of disodium
salt of 9-(o-carboxyphenyl)-6-hydroxy2,4,5,7-tetraiodo-3-isoxanthone
monohydrate and subsidiary colouring matters together with water, sodium
chloride and/or sodium sulfate as the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
Disodium
salt of 9-(o-carboxyphenyl)-6-hydroxy-2,4,5,7-tetraiodo-3- isoxanthone
monohydrate
C.A.S.
number
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Chemical
formula
C20H6I4Na2O5.H2O
Structural
formula
Formula
weight
897.88
3. Description
Red
powder or granules.
4. Functional
uses
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5. Specifications
5.1. Identification
Solubility
Soluble
in water, slightly soluble in ethanol
Identification
of colouring matters
Passes
test.
5.2. Purity
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Not more
than 13% together with chloride and sulfate calculated as sodium salts.
Inorganic
iodides
Not more
than 0.1% calculated as sodium iodide.
Water
insoluble matter
Not more
than 0.2%.
Zinc
Not more
than 50 mg/kg.
Lead
Not more than 2 mg/kg.
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Not more than 4% (except
fluorescein).
Fluorescein
Not more
than 20 mg/kg.
Organic
compounds other than colouring matters
Tri-iodoresorcinol:
Not more than 0.2%
2-(2,4-dihydroxy-3,5-di-iodobenzoyl)
benzoic acid: Not more than 0.2%
Ether
extractable matter
From a
solution of pH not less than 7, not more than 0.2%.
5.3. Content
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6. Tests
6.1.
Purity
Inorganic
iodides
Weigh 1.0 g of the sample into a
100-ml beaker. Add 75 ml distilled water and the magnetic follower. Stir to
dissolve. Immerse a iodide specific electrode and a reference electrode in
the solution and set a suitable millivoltmeter to read the potential of the
system in millivolts.
Add 0.001 M silver nitrate
solution from a burette initially in 0.5 ml aliquots, reducing these to 0.1
ml as the end-point approaches as indicated by an increasing change in
potential for each addition. After allowing time for the reading to
stabilize, record the millivolt readings after each addition. Continue the
titration until further additions make little change in the potential. Plot
the millivolt readings against the volume of silver nitrate solution added. The
equivalent point is the volume corresponding to the maximum slope of the
curve.
The percentage of sodium iodide
in sample is: Titre x 0.015%.
Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
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Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 5; Height of ascent of solvent front: approximately
17 cm. Note: Take special care not to allow the chromatograms to be
exposed to direct sunlight.
Organic
compounds other than colouring matters
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Proceed
as directed under Column chromatography, using, for example, the following
absorptivities:
Tri-iodoresorcinol:
0.079 mg L-1cm-1 at 233 nm (acidic).
2(2,4-dihydroxy-3,5-di-iodobenzoyl)benzoic
acid: 0.047 mg L-1cm-1 at 348 nm (alkaline).
Fluorescein
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The fluorescein is separated from
the sample by TLC and compared with a standard chromatogram prepared from
fluorescein at the concentration corresponding to the limit figure.
Solvent:
Methanol+water+ammonia (s.g.
0.890) (500 ml+400 ml+100 ml).
Sample:
Weigh 1.0 g of the sample,
dissolve in about 50 ml solvent and dilute to 100 ml in a volumetric flask.
Standard:
Weigh an amount of fluorescein,
previously purified by recrystallisation from ethanol, equal to 1 g x the
colouring matter content of the sample as determined under Assay. Dissolve
in water (or in water with 10 ml ammonia s.g. 0.890 if fluorescein-free acid
is being used) and dilute to 100 ml. Make further sequential dilutions as
follows:
1 ml to 100 ml with water.
1 ml to 100 ml with water.
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Chromatography solvent:
n-Butanol+water+ammonia (s.g.
0.890)+ethanol (100 ml+44 ml+1 ml+ 22.5 ml).
Procedure:
Spot 25 µl of the sample and
standard solutions side by side on a cellulose plate. Develop for 16 h in the
chromatography solvent. Allow the plate to dry. View under a UV light source
and compare the fluorescence of the standard with the fluorescence of the
corresponding area on the chromatogram of the sample. The intensity of the
latter shall not be greater than that of the former.
Note: Take special care
not to allow the chromatograms to be exposed to direct sunlight.
6.2.
Method of assay
Dissolve about 1 g of the sample,
accurately weighed, in 250 ml of water, transfer to a clean 500-ml beaker,
add 8.0 ml of 1.5 N nitric acid and stir well. Filter through a sintered
glass crucible (porosity 3, diameter 5 cm) which has been weighed containing
a small glass stirring rod. Wash thoroughly with 0.5% nitric acid until the
filtrate gives no turbidity with silver nitrate TS, and then wash with 30 ml
water. Dry to constant weight at 135±5º, carefully breaking up the
precipitate by means of the glass rod. Cool in a desiccator and weigh
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Determination of Hydrochloric
Acid-insoluble Matter in Erythrosine Lake
Reagents:
Concentrated hydrochloric acid
Hydrochloric acid 0.5% v/v
Dilute ammonia solution (dilute
10 ml ammonia, s.g. 0.890 to 100 ml with water).
Procedure:
Accurately weigh approximately 5
g of the lake into a 500-ml beaker. Add 250 ml water and 60 ml concentrated
hydrochloric acid. Boil to dissolve the alumina while the Erythrosine
converts to its "free acid" form, which is insoluble in acid. Filter
through a tared No. 4 sintered glass crucible. Wash the crucible with a small
amount of hot 0.5% hydrochloric acid and then with some hot distilled water. Remove
the acid filtrate from the filter flask, replace the crucible and wash with
hot dilute ammonia solution until the washings are colourless. Dry the
crucible to constant weight at 135º. Express the residue as a percentage of
the weight taken.
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ANNEX 11
SPECIFICATIONS AND TESTS FOR RED 2G
1. Synonyms
CI Food
Red 10, Azogeranine; CI (1975): 18050
INS: 128
ADI = 0 -
1 mg/kg bw.
2. Definition
Consists essentially of disodium
8-acetamido-1-hydroxy-2-phenylazo3,6-naphthalenedisulfonate and subsidiary
colouring matters together with sodium chloride and/or sodium sulfate as the
principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
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Disodium
8-acetamido-1-hydroxy-2-phenylazo-3,6-naphthalenedisulfonate
C.A.S.
number
3734-67-6
Chemical
formula
C18H13N3Na2O8S2
Structural
formula
Formula
weight
509.43
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Red
powder or granules.
4. Functional
uses
Colour.
5. Specifications
5.1. Identification
Solubility
Freely
soluble in water; sparingly soluble in ethanol.
Identification
of colouring matters
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5.2. Purity
Loss on
drying at 135 °C
Not more
than 20% together with chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
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Organic
compounds other than colouring matters
Not more than 0.3% of sum of
5-acetamido-4-hydroxy-2,7-naphthalene2,7-disulfonic acid and 5-Amino-4-hydroxy-2,7-naphthalene-2,7-
disulfonic acid.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
Ether
extractable matter
Not more
than 0.2%.
5.3. Content
Not less
than 80% total colouring matter.
6. Tests
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Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 4; Height of ascent of solvent front: approximately
17 cm.
Organic
compounds other than colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
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6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 0.6 - 0.7 g;
Buffer:
15 g sodium hydrogen tartrate;
Weight: (D) of colouring matters
equivalent to 1.00 ml of 0.1 N TiCl3: 12,74 mg
ANNEX 12
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1. Synonyms
CI Food
Red 17, FD&C Red No. 40; CI (1975): 16035
INS: 129
ADI = 0 -
7 mg/kg bw.
2. Definition
Consists of disodium
6-hydroxy-5-(2-methoxy-5-methyl-4- sulfonatophenylazo)-2-naphthalenesulfonate
and subsidiary colouring matters together with sodium chloride and/or sodium
sulfate as the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
Disodium
6-hydroxy-5-(2-methoxy-5-methyl-4-sulfonatophenylazo)-2- naphthalenesulfonate
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25956-17-6
Chemical
formula
C18H14N2Na2O8S2
Structural
formula
Formula
weight
496.43
3. Description
Dark red
powder or granules.
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Colour.
5. Specifications
5.1. Identification
Solubility
Soluble in water; sparingly
soluble in ethanol.
Identification
of colouring matters
Passes test.
5.2. Purity
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Loss on
drying at 135 °C
Not more than 15% together with
chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
Not more than 3%.
Organic
compounds other than colouring matters
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Not more than 0.2% of
4-amino-5-methoxy-2-methyl-benzene-sulfonic acid;
Not more than 1.0% of disodium
6,6'-oxybis(2-naphthalene-sulfonate);
Unsulfonated primary
aromatic amines
Not more than 0.01% calculated as
aniline.
Ether
extractable matter
Not more than 0.2%.
5.3. Content
Not less
than 85% total colouring matters
6. Tests
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Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 4; Height of ascent of solvent front: approximately
17 cm.
Organic
compounds other than colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
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6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 0.5 - 0.6 g;
Buffer:
15 g sodium hydrogen tartrate;
Weight
(D) of colouring matters equivalent to 1.00 ml of 0.1 N TiCl3:
12.41 mg.
ANNEX 13
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1. Synonyms
Indigocarmin;
CI Food Blue 1, FD&C Blue No. 2; CI (1975): 73015
INS: 132
ADI = 0 -
5 mg/kg bw.
2. Definition
Consists essentially of a mixture
of disodium 3,3' -dioxo-[delta2,2'- biindoline]-5,5'-disulfonate
(principal component) and disodium 3,3'- dioxo-[delta2,2'-biindoline]-5,7'-disulfonate
(isomer) and subsidiary colouring matters together with NaCl and/or Na2SO4 as
the principal uncoloured components.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
Disodium 3,3'-dioxo-[delta2,2'-biindoline]-5,5'-disulfonate
(principal component)
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860-22-0
(5,5' isomer)
Chemical
formula
C16H8N2Na2O8S2
Structural
formula
Formula
weight
466.36
3. Description
Blue
powder or granules.
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Colour.
5. Specifications
5.1. Identification
Solubility
Soluble in water; sparingly
soluble in ethanol.
Identification
of colouring matters
Passes test.
5.2. Purity
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Loss on
drying at 135 °C
Not more than 15% together with
chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more than 0.2%.
Lead
Not more than 2 mg/kg.
Subsidiary
colouring matters
Not more than 1% (except disodium
3,3'-dioxo-[delta2,2'-biindoline]- 5,7'- disulfonate).
Organic
compounds other than colouring matters
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Unsulfonated primary
aromatic amines
Not more than 0.01% calculated as
aniline.
Ether
extractable matter
Not more
than 0.2%.
5.3. Content
Not less
than 85% total colouring matters.
Not more
than 18% of disodium 3,3'-dioxo-[delta2,2'-biindoline]-5,7'-
disulfonate
6. Tests
6.1.
Purity
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Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 3; Height of ascent of solvent front: approximately
17 cm.
Note 1. The 5,7' isomer is
separated as a wide blue zone just in front of the main blue band. Do not
include this zone in the subsidiary colouring matter zones which are cut out
and measured.
Note 2. The 15 ml sodium
hydrogen carbonate solution used in the general procedure is replaced by 15
ml 0.05 N hydrochloric acid in order to avoid the decomposition which the
sulfonated indigo undergoes in alkaline solution.
Organic
compounds other than colouring matters
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- Use liquid chromatography under
the following conditions: HPLC elution gradient: 2 to 100% gradient followed
by elution at 100%.
Ether
extractable matter
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Weigh
accurately about 2 g sample instead of the 5 g stated in the general methods.
6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 1.0 - 1.1 g;
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Weight (D) of colouring matters
equivalent to 1.00 ml of 0.1 N TiCl3: 23.32 mg.
Isomer
content by paper chromatography:
Refer to the conditions for the
determination of subsidiary colouring matters. Cut the isomer band from the
chromatogram in the manner detailed for the subsidiary bands, extract into
solvent and measure the absorbance at its Smax. Measure the absorbance of the
corresponding blank at the same wavelength. As a standard use 0.1 ml of an
0.20% solution of the sample applied to the 18 cm x 0.7 cm rectangle.
Isomer expressed as a percentage
of the sample
% = [A /
As] x 20% x [D / 100]
where:
A is the net absorbances (of the
corresponding blank) of the isomer.
As is the net
absorbances (of the corresponding blank) of the standard.
D is the total colouring matters
content of the sample.
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The 5,7' isomer separates under
the HPLC conditions detailed above for the separation of subsidiary colouring
matters, and the amount present can be quantified.
ANNEX 14
SPECIFICATIONS AND TESTS FOR SUNSET BRILLIANT BLUE FCF
1. Synonyms
CI Food
Blue 2, FD&C Blue No. 1; CI (1975): 42900
INS: 133
ADI = 0 -
1 mg/kg bw.
2. Definition
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May be converted to the corresponding
aluminium lake in which case only the General Specifications for Aluminium
Lakes of Colouring Matters applies.
Chemical
names
Disodium
3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-sulfonatobenzyl)-
amino]phenyl](2-sulfonatophenyl)methylene]-2,5-cyclohexa-diene-1-
ylidene]ammoniomethyl]-benzenesulfonate.
Disodium
I-[4-(N-ethyl-3-sulfonatobenzylamino)phenyl]-I- [4-(N-ethyl-3-
sulfonatobenzyliminio)cyclohexa-2,5-dienyli-dene]toluene-2-sulfonate (an
alternative chemical name).
C.A.S.
number
3844-45-9
Chemical
formula
C37H34N2Na2O9S3
Structural
formula
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Formula
weight
792.86
3. Description
Blue
powder or granules.
4. Functional
uses
Colour.
5. Specifications
5.1. Identification
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Soluble in water; sparingly
soluble in ethanol.
Identification
of colouring matters
Passes test.
5.2. Purity
Loss on
drying at 135 °C
Not more than 15% together with
chloride and sulfate calculated as sodium salts.
Water
insoluble matter
Not more than 0.2%.
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Not more than 2 mg/kg.
Chromium
Not more than 50 mg/kg.
Subsidiary
colouring matters
Not more than 6%.
Organic
compounds other than colouring matters
Not more than 1.5%, sum of 2-, 3-
and 4-formylbenzenesulfonic acids.
Not more tan 0.3%
3-[[N-ethyl-N-(4-sulfophenyl) amino] methyl] benzenesulfonic acid.
Leuco
base
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Unsulfonated primary
aromatic amines
Not more than 0.01% calculated as
aniline.
Ether
extractable matter
Not more than 0.2%.
5.3. Content
Not less
than 85% total colouring matter.
6. Tests
6.1.
Purity
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- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 4; Develop chromatogram for approximately 20 hours.
Organic
compounds other than colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Proceed as directed under
Column chromatography, using, for example, the following absorptivities:
3-formylbenzenesulfonic acid:
0.0495 mgL-1/cm-1 at 246 nm in dilute HCl.
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Leuco
base
Tested as directed under JECFA
monograph 1-Vol. 4. Weigh accurately 120±5 mg sample and proceed as directed
under Leuco Base in Sulfonated Triarylmethane Colours (Vol. 4) Absorptivity
(a) = 0.164 mgL-1cm-1 at 629 nm Ratio = 0.9706.
6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
Weight of
sample: 1.8 - 1.9 g;
Buffer:
15 g sodium hydrogen tartrate;
Weight
(D) of colouring matters equivalent to 1.00 ml of 0.1 N TiCl3:
39.65 mg.
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ANNEX 15
SPECIFICATIONS AND TESTS FOR CHLOROPHYLLS
1. Synonyms
Magnesium chlorophyll, magnesium
phaeophytin;
Clorophyllin; C.I. Natural Green
3; C.I. (1975): 75810;
INS 140
ADI “Not limited”.
2. Definition
Obtained by solvent extraction of
grass, lucerne, nettle and other plant material; during the subsequent
removal of solvent, the naturally present coordinated magnesium may be wholly
or partly removed from the chlorophylls to give the corresponding
phaeophytins; the principal colouring matters are the phaeophytins and
magnesium chlorophylls. The extracted product, from which the solvent has
been removed, contains other pigments such as carotenoids as well as oils,
fats and waxes derived from the source material. Only the following solvents
may be used for the extraction: acetone, dichloromethane, methanol, ethanol,
propan-2-ol and hexane.
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The major colouring principles
are:
Phytyl
(132R,17S,18S)-3-(8-ethyl-132-methoxycarbonyl-2,7,12,18-tetramethyl13'-oxo-3-vinyl-131
-132 -17,18-tetrahydrocyclopenta [at]-prophylrin-17-
yl)propionate, (Pheophytin a), or as the magnesium complex (Chlorophyll a).
Phytyl
(132R,17S,18S)-3-(8-ethyl-7-formyl-132 -methoxycarbonyl-2,12,18-
trimethyl-13'-oxo-3-vinyl-131 -132 -17,18-tetrahydro-cyclopenta
[at]-prophylrin17-yl)propionate, (Pheophytin b), or as the magnesium complex
(Chlorophyll b).
C.A.S. number
Phaeophytin a, Magnesium complex:
479-61-8
Phaeophytin b, Magnesium complex:
519-62-0
Chemical formula
Phaeophytin a Magnesium complex
(Chlorophyll a): C55H72MgN4O5
Phaeophytin a: C55H74N4O5
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Phaeophytin b: C55H72N4O6
Structural formula
where:
X = CH3
for the "a" series
X = CHO
for the "b" series.
Loss of Mg converts chlorophyll
into the corresponding phaeophytin
Formula weight
Phaeophytin a magnesium complex
(Chlorophyll a): 893.51
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Phaeophytin b magnesium complex
(Chlorophyll b): 907.49
Phaeophytin: 885.20
3. Description
Waxy solid ranging in colour from
olive green to dark green depending on the content of co-ordinated magnesium.
4. Functional uses
Colour.
5. Specifications
5.1. Identification
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Insoluble in water; soluble in
ethanol, diethyl ether, chloroalkanes, hydrocarbons and fixed oils.
Thin-layer chromatography
Passes test (described in
“Tests”).
5.2. Purity
Residual solvents
Acetone, methanol, ethanol,
propan-2-ol, hexane: Not more than 50 mg/kg, singly or in combination.
Dichloromethane:
Not more than 10 mg/kg
Arsenic
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Not more than 3 mg/kg.
(Tested as directed under JECFA
monograph 1-Vol. 4 - Method II).
Lead
Not more than 5 mg/kg.
(Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
5.3. Content
Content of total combined
phaeophytins and their magnesium complexes is not less than 10%.
6. Tests
6.1. Identification
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Thin-layer chromatography
Apply a 1 in 20 solution of the
sample in chloroform as a band of the length of 2 cm to a Silica 60 TLC
plate. After drying, develop the plate by a mixture of 50% hexane, 45%
chloroform and 5% ethanol (General purpose reagent grade chloroform is
supplied with 2% of added ethanol as a stabilizer.
The 5% ethanol in the solvent
mixture is in addition to this), until the solvent ascends to a point 15 cm
above the initial spots. Allow the solvent to evaporate, then visually
examine the separated spots and identify the components of interests by their
Rf values and colours. Approximate Rf values and colour
of the spots are as follows:
Phaeophytin a: 0.77; grey/brown
Phaeophytin b: 0.75; yellow/brown
Chlorophyll a: 0.50; blue/green
Chlorophyll b: 0.63; yellow/green
In addition spots may be visible
for b-carotene
at Rf 0.81 and xanthophyll at Rf 0.47 and 0.23.
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Residual solvents
Tested as directed under JECFA
monograph 1-Vol. 4.
Determine by gas
chromatographically using either the method of entrainment distillation (Determination
of Residual Solvents) or headspace analysis (Limit Test for Solvent
Residues).
Arsenic
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Method II.
Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
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6.1. Method of assay
Accurately weigh about 100 mg of
the sample and dissolve in diethyl ether, making the volume to 100 ml. Dilute
2 ml of this solution to 25 ml with diethyl ether. The concentration of the
sample should not give an absorbance at 660.4 nm that is in excess of the
working range for Absorbance measurements, i.e., not in excess of 0.7.
Measure the absorbances of the
solution in a 1 cm cell against a diethyl ether blank at 660.4 nm, 642.0 nm,
667.2 nm and 654.4 nm. (These being the absorbance maxima in diethyl ether
for chlorophyll a, chlorophyll b, phaeophytin a, and phaeophytin b,
respectively). In addition measure at 649.8 nm and 628.2 nm. To the remaining
diluted solution add one crystal of oxalic acid and after dissolution and
mixing, remeasure the absorbances at the same wavelengths. "dA"
is the difference between the absorbances between the absorbance at the
respective wavelengths, before and after addition of oxalic acid.
Calculate the concentration of
the individual compounds in mmol/L from the following equations:
Chlorophyll a = 17.7 d A (660.4
nm) + 7.15 d A (642.0 nm) Chlorophyll b = 19.4 d A (642.0
nm) - 2.92 d A (660.4 nm) Phaeophytin a = -4.89 A (649.8 nm) + 0.0549 A
(628.2 nm) +18.7 A (667.2 nm) + 0.0575 A (654.4 nm) - chlorophyll a
Phaeophytin b = -71.0 A (649.8 nm) + 2.51 A (628.2 nm) - 13.5 A (667.2 nm) +
84.3 A (654.4 nm) - chlorophyll b
Convert the figures in mmol/L to
percentages using the following equations:
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ANNEX 16
SPECIFICATIONS AND TESTS FOR CHLOROPHYLLS, COPPER COMPLEXES
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Copper chlorophyllin, copper
phaeophytin
C.I. Natural Green 3; C.I. (1975)
No. 75810;
INS: 141i
ADI = 0 - 15 mg/kg bw.
2. Definition
Obtained by addition of an
organic salt of copper to the substance obtained by solvent extraction of
grass, lucerne, nettle and other plant material; the product, from which the
solvent has been removed, contains other pigments such as carotenoids as well
as fats and waxes derived from the source material. The principal colouring
matters are the copper phaeophytins. Only the following solvents may be used
for the extraction: acetone, dichloromethane, methanol, ethanol, propan-2-ol
and hexane.
Chemical names
The major colouring principles
are:
[Phytyl
(132R,17S,18S)-3-(8-ethyl-132
-methoxycarbonyl-2,7,12,18- tetramethyl-131 -oxo-3-vinyl-131 ,132
,17,18-tetra-hydrocyclopenta [at]- prophyrin-17-yl)propionate] copper (II)
(Copper chlorophyll a)
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C.A.S. number
65963-40-8
Chemical formula
Copper phaeophytin a: C55H72CuN4O5
Copper phaeophytin b: C55H70CuN4O6
Structural formula
where:
X = CH3
for the "a" compound
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Formula weight
Copper phaeophytin a: 932.75
Copper phaeophytin b: 946.73
3. Description
Waxy solid ranging in colour from
blue green to dark green depending on the source material.
4. Functional uses
Colour.
5. Characteristics
5.1. Identification
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Solubility
Insoluble in water; soluble in
ethanol, diethyl ether, chloroalkanes, hydrocarbons and fixed oils.
Spectrophotometry
A (1%, 1 cm) at 405 nm in
chloroform is not less than 54.
Thin-layer chromatography
Passes test (described in
“Tests”).
5.2. Purity
Residual solvents
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Dichloromethane:
Not more than 10 mg/kg.
Free ionizable copper
Not more than 200 mg/kg.
Total copper
Not more than 8% of the total
copper phaeophytins.
Arsenic
Not more than 3 mg/kg.
Lead
Not more than 5 mg/kg.
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Not less than 10% of total copper
phaeophytins.
6. Tests
5.1. Identification
Thin-layer chromatography
Apply a 1 in 20 solution of the
sample in chloroform as a band of the length of 2 cm to a Silica 60 TLC
plate. After drying, develop the plate by a mixture of 50% hexane, 45%
chloroform and 5% ethanol (General purpose reagent grade chloroform is
supplied with 2% of added ethanol as a stabilizer. The 5% ethanol in the
solvent mixture is in addition to this), until the solvent ascends to a point
15 cm above the initial spots. Allow the solvent to evaporate, then visually
examine the separated spots and identify the components of interests by their
Rf values and colours. Approximate Rf values and colour
of the spots are as follows:
Copper phaeophytin a: 0.5; green
Copper phaeophytin b: 0.73;
yellow/green
In addition spots may be visible
for b-carotene
at Rf 0.81 and xanthophyll at Rf 0.47 and 0.23.
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Residual solvents
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine by gas
chromatographically using either the method of entrainment distillation (Determination
of Residual Solvents) or headspace analysis (Limit Test for Solvent
Residues).
Free ionizable copper
Accurately weigh about 1 g of the
sample and dissolve in 20 ml of arachid oil, with the aid of gentle heat. Add
exactly 200 ml of water, stir mechanically, and adjust to pH 3.0 by careful
addition of 0.5 N hydrochloric acid (avoid overshooting). Allow the mixture
to stand for 10 min. If necessary readjust to pH 3.0 by careful addition of
0.5 N hydrochloric acid. Transfer to a separating funnel and allow to stand
for about 20 min. Filter the aqueous phase through a No. 50 Whatman filter
paper, rejecting the first 10 ml.Subject this solution to analysis for copper
by atomic absorption spectrometry - JECFA monograph 1-Vol. 4.
Total copper
Ignite about 0.1 g, accurately
weighed, of the sample contained in a silica dish, at a temperature not
exceeding 500º, until all carbon is removed; moisten with one or two drops of
concentrated sulphuric acid and re-ash. Dissolve the ash by boiling with 3
portions (each of 5 ml) of 10% (w/w) hydrochloric acid, filtering each
addition through the same small filter paper into a 100 ml volumetric flask. Cool,
and make up to volume with purified water. Subject this solution to analysis
for copper by atomic absorption spectrometry - See JECFA monograph
1-Vol. 4.
Arsenic
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- Method II.
Lead
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
6.3. Method of assay
Accurately weigh about 100 mg of
the sample and dissolve in diethyl ether, making the volume to 100 ml. Dilute
2 ml of this solution to 25 ml with diethyl ether. The concentration of the
sample should not give an absorbance at 660.4 nm that is in excess of the
working range for Absorbance measurements, i.e., not in excess of 0.7.
Measure the absorbances (A) of
the solution in a 1 cm cell against a diethyl ether blank at 667.2 nm, 654.4
nm, 649.8 nm and 628.2 nm. (The latter two wavelengths being the absorbance
maxima in diethyl ether for copper phaeophytin a and copper phaeophytin b
respectively).
Calculate the concentration of the
individual compounds in (mol/L from the following equations:
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Copper phaeophytin b = -8.46 A
(649.8nm) + 20.7 A (628.2nm) - 1.69 A (667.2nm) + 5.13 A (654.4nm)
Convert the figures in mmol/L to
percentages using the following equations:
ANNEX 17
SPECIFICATIONS AND TESTS FOR CHLOROPHYLLINS, COPPER COMPLEXES
SODIUM AND POTASSIUM SALTS
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Sodium copper chlorophyllin;
potassium copper chlorophyllin
Chlorophyllins, C.I. (1975) No.
75810;
INS 141ii
ADI = 0 - -15 mg/kg bw.
2. Definition
The alkali salts of Copper
Chlorophyllins are obtained by the addition copper to the product obtained by
the saponification of a solvent extraction of grass, lucerne, nettle and
other plant material; the saponification removes the methyl and cyclophytol
ester groups and may partially cleave the pentenyl ring; after addition of
copper to the purified chlorophyllins, the acid groups are neutralised to
form the salts of potassium and/or sodium; the commercial products may be
presented as aqueous solutions or dried powders. Only the following solvents
may be used for the extraction: acetone, dichloromethane, methanol, ethanol,
propan-2-ol and hexane.
Chemical names
The major colouring principles
are:
3-(10-Carboxylato-4-ethyl-1,3,5,8-tetramethyl-9-oxo-2-vinylphorbin-7-
yl)propionate, copper complex (Copper chlorophyllin a)
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Depending on the degree of
hydrolysis the cyclopentenyl ring may be cleaved with the resultant
production of a third carboxyl function.
Chemical formula
Copper phaeophytin a: C34H32CuN4O5
Copper chlorophyllin b (acid form):
C34H30CuN4O6
Structural formula
where:
X = CH3
for the "a" series
X = CHO
for the "b" compound.
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Formula weight
Copper
chlorophyllin a: 640.20
Copper
chlorophyllin b: 654.18
Each may be increased by a 18
Daltons if the cyclopentenyl ring is cleaved.
3. Description
Dark green to blue/black powder
or dark green solution.
4. Functional uses
Colour.
5. Specifications
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Solubility
Soluble in water; very slightly
soluble in lower alcohols and ketones and diethyl ether; insoluble in
chloroalkanes, hydrocarbons and fixed oils.
Spectrophotometry
Passes test (described in
“Tests”).
Test for copper
Passes test (described in
“Tests”).
Test for sodium
Passes test.
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Passes test.
5.2. Purity
Basic
dyes
Passes test (described in
“Tests”).
Residual solvents
Acetone, methanol, ethanol,
propan-2-ol, hexane: Not more than 50 mg/kg, singly or in combination.
Dichloromethane: Not more than 10
mg/kg
Free ionizable copper
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Total copper
Not more than 8% of the total
copper phaeophytins.
Arsenic
Not more than 3 mg/kg.
Lead
Not more than 5 mg/kg.
5.3. Content
Not less than 95% total copper
chlorophyllins after drying (100º, 1 h).
6. Tests
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Spectrophotometry
A (1%, 1 cm) of the sample, dried
at 100º for 1 h, at 405 nm in pH 7.5 phosphate buffer is not less than 540.
Test for copper
Dissolve the sulfated ash of the
sample (using 1 g of sample, Method I) in 10 ml of dilute hydrochloric acid
TS by heating on a water bath. Filter if the solution is not clear, and
dilute to 10 ml with water. Use this solution as the test preparation for the
following tests:
To 5 ml of the test preparation
add ammonia TS to make the solution alkaline. Blue colour should appear.
To 5 ml of the above test
preparation add 0.5 ml of a 1 in 1000 sodium diethyldithiocarbamate solution.
A brown precipitate should be formed.
Test for sodium
Tested as directed under JECFA
monograph 1-Vol. 4 - Test the solution described under Tests, identification
tests Test for copper.
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Tested as directed under JECFA
monograph 1-Vol. 4 - Test the solution described under Tests, identification
tests Test for copper.
6.2 Purity
Basic
dyes
To 5 ml of a 0.5% aqueous solution
of the sample in a test- tube add 1 ml of 1 N hydrochloric acid and add 5 ml
of diethyl ether. Mix thoroughly and allow to separate. The ether layer
should be no darker than pale green.
Residual solvents
Determine by gas
chromatographically using either the method of entrainment distillation (Determination
of Residual Solvents) or headspace analysis (Limit Test for Solvent
Residues). (Tested as directed under JECFA monograph 1-Vol. 4)
Free ionizable copper
Accurately weigh about 1 g of the
sample and mix in 20 ml of arachid oil, with the aid of gentle heat. Add
exactly 200 ml of water, stir mechanically, and adjust to pH 3.0 by careful
addition of 0.5 N hydrochloric acid (avoid overshooting). Allow the mixture
to stand for 10 min. If necessary readjust to pH 3.0 by careful addition of
0.5 N hydrochloric acid. Transfer to a separating funnel and allow to stand
for about 20 min. Filter the aqueous phase through a No. 50 Whatman filter
paper, rejecting the first 10 ml. Subject this solution to analysis for
copper by atomic absorption spectrometry (see JECFA monograph 1-Vol.
4).
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Ignite about 0.1 g, accurately
weighed, of the sample contained in a silica dish, at a temperature not
exceeding 500º, until all carbon is removed; moisten with one or two drops of
concentrated sulphuric acid and re-ash. Dissolve the ash by boiling with 3
portions (each of 5 ml) of 10% (w/w) hydrochloric acid, filtering each
addition through the same small filter paper into a 100 ml volumetric flask. Cool,
and make up to volume with purified water. Subject this solution to analysis
for copper by atomic absorption spectrometry (see JECFA monograph
1-Vol. 4).
Arsenic
Tested as directed under JECFA
monograph 1-Vol. 4- Method II
Lead
Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph 1-Vol. 4, “Instrumental Methods.”
6.3. Method of assay
Weigh accurately about 1 g of the
sample, dried previously at 100º for 1 h, then dissolve in 20 ml Phosphate
Buffer Solution (pH 7.5) and dilute to 1000 ml with distilled water. Dilute
10 ml of this solution to 100 ml with Phosphate Buffer Solution (pH 7.5).
Measure the optical density of the final solution (0.001% w/v) in a suitable
spectrophotometer, using a 1 cm cell and slit width of 0.10 mm at 403-406 nm,
recording the maximum within this range.
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Optical density ´ 104
--------------------------------------
565 ´ weight of sample (g)
This formula was derived on the
assumption that 100% pure sodium copper chlorophyllin has a specific
absorbance of 565.
ANNEX 18
SPECIFICATIONS AND TESTS FOR GREEN S
1. Synonyms
CI food
green 4, food green S; CI (1975): 44090
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ADI “Not specified”.
2. Definition
Consists
essentially of sodium N-[4-[[4-(dimethylamino)phenyl](2-
hydroxy-3,6-disulfo-1-naphthalenyl)-methylene]-2,5-cyclohexadien-1-
ylidene]-N-methylmethanaminium and subsidiary colouring matters together with
sodium chloride and/or sodium sulfate as the principal uncoloured compounds.
May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies.
Chemical
names
Sodium
N-[4-[[4-(dimethylamino)phenyl](2-hydroxy-3,6-disulfo-1-
naphtalenyl)-methylene]-2,5-cyclohexadien-1-ylidene]-N-methylmethanaminium;
or Sodium
5-[4-dimethylamino-I-(4-dimethyliminiocyclohexa-2,5- dienylidene)
benzyl]-6-hydroxy-7-sulfonato-naphthalene-2-sulfonate.
C.A.S.
number
860-22-0
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C27H25N2NaO7S2
Công thức
cấu tạo
Formula
weight
576.63
3. Description
Dark
green powder or granules.
4. Functional
uses
Colour.
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5.1. Identification
Solubility
Soluble
in water; slightly soluble in ethanol.
Identification
of colouring matters
Passes
test.
5.2. Purity
Loss on
drying at 135 °C
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Water
insoluble matter
Not more
than 0.2%.
Lead
Not more than 2 mg/kg.
Chromium
Not more
than 50 mg/kg.
Subsidiary
colouring matters
Not more than 1%.
Organic
compounds other than colouring matters
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Not more
than 0.1% of 4,4'-Bis (dimethylamino) benzophenone.
Not more than 0.2% of
3-Hydroxynaphthalene-2,7-disulfonic acid.
Unsulfonated primary
aromatic amines
Not more
than 0.01% calculated as aniline.
Leuco
base
Not more than 5.0%.
Ether
extractable matter
Not more
than 0.2%.
5.3. Content
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6. Tests
6.1. Purity
Lead
- Tested
as directed under JECFA monograph 1-Vol. 4.
- Determine
using an atomic absorption technique appropriate to the specified level. The
selection of sample size and method of sample preparation may be based on the
principles of the method described in JECFA monograph 1-Vol. 4, “Instrumental
Methods.”
Subsidiary
colouring matters
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use the following conditions:
Developing solvent: No. 2; Height of ascent of solvent front: approximately
17 cm.
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- Tested as directed under JECFA
monograph 1-Vol. 4.
- Use liquid chromatography under
the following conditions: HPLC elution gradient: 2 to 100% at 2% per min.
Leuco
base
- Tested as directed under JECFA
monograph 1-Vol. 4.
- Weigh accurately 110±5 mg of
sample and proceed as directed under Leuco Base in Sulfonated Triarylmethane
Colours. Absorptivity: (a) = 0.1725 mg L-1 cm-1 at
approximately 634 nm. Ratio = 0.9600
6.2.
Method of assay
Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following:
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Buffer:
15 g sodium hydrogen tartrate;
Weight
(D) of colouring matters equivalent to 1.00 ml 0.1 N TiCl3: 28.83 mg.
ANNEX 19
SPECIFICATIONS AND TESTS FOR CARAMEL
1. Synonyms
Caramel
colours are divided into four classes. The synonyms for each class are:
Class I:
Plain caramel, caustic caramel; INS No. 150a
Class
III: Ammonia caramel; INS No. 150c
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ADI for
class I “Not specified”;
ADI for
class III = 0-160 mg/kg bw
ADI for
class IV = 0-200 mg/kg bw (0-150 mg/kg on solids basis)
2. Definition
Complex
mixtures of compounds, some of which are in the form of colloidal aggregates,
manufactured by heating carbohydrates either alone or in the presence of
food-grade acids, alkalis or salts; classified according to the reactants
used in their manufacture as follows:
Class I:
Prepared by heating carbohydrates with or without acids or alkalis; no
ammonium or sulfite compounds are used.
Class
III: Prepared by heating carbohydrates with or without acids or alkalis in
the presence of ammonium compounds; no sulfite compounds are used.
Class IV:
Prepared by heating carbohydrates with or without acids or alkalis in the
presence of both sulfite and ammonium compounds.
In all
cases the carbohydrate raw materials are commercially available foodgrade
nutritive sweeteners consisting of glucose, fructose and/or polymers thereof. The
acids and alkalis are food-grade sulfuric or citric acids and sodium, potassium
or calcium hydroxides or mixtures thereof.
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Where
sulfite compounds are used they are one or any of the following: sulfurous
acid, potassium, sodium and ammonium sulfites and hydrogen sulfites.
Food-grade
anti-foaming agents may be used as processing aids during manufacture.
3. Description
Dark
brown to black liquids or solids having an odour of burnt sugar.
4. Functional
uses
Colour
5. Specifications
5.1. Identification
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Miscible
with water
Identification
of colouring matters
Passes
test.
Classification
Class I:
Not more than 50% of the colour is bound by DEAE Cellulose and not more than
50% of the colour is bound by Phosphoryl Cellulose.
Class
III: Not more than 50% of the colour is bound by DEAE Cellulose and more than
50% of the colour is bound by Phosphoryl Cellulose.
Class IV:
More than 50% of the colour is bound by DEAE Cellulose and it exhibits an
Absorbance Ratio of not more than 50.
See
description under TESTS.
5.2. Purity
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Solid
content
Class I:
62-77%
Class
III: 53-83%
Class IV:
40-75%
Colour
intensity
Class I:
0.01-0.12
Class
III: 0.08-0.36
Class IV:
0.10-0.60
Total
nitrogen
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Class
III: 1.3 -6.8%
Class IV:
0.5-7.5%
Total
sulfur
Class I:
max 0.3%
Class
III: max 0.3%
Class IV:
1.4-10.0%
Sulfur
dioxide
Class I:
-
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Class IV:
max 0.5%
Ammoniacal
nitrogen
Class I:
-
Class
III: max 0.4%
Class IV:
max 2.8%
4-Methylimidazole
(MEI)
Class I:
-
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Class IV:
max 1000 mg/kg & max 250 mg/kg on an equivalent colour basis
2-Acetyl-4-tetrahydroxybutylimidazole
(THI)
Class I:
-
Class
III: max 40 mg/kg & max 25 mg/kg on an equivalent colour basis.
Class IV:
-
Arsenic
Not more
than 1 mg/kg
Lead
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6. Tests
6.1. Identification
Classification/Colour
bound by DEAE Cellulose
For the purposes of this
specification, colour bound by DEAE cellulose is defined as the percentage of
decrease in absorbance of a caramel colour solution at 560 nm after treatment
with DEAE Cellulose.
Special reagent:
DEAE (diethylaminoethyl)
Cellulose of 0.7 meq/gram capacity, e.g. Cellex D from Bio-Rad or equivalent
DEAE Celluloses of higher or lower capacities in proportionately higher or
lower quantities.
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Prepare a caramel colour solution
of approximately 0.5 absorbance unit at 560 nm by transferring an appropriate
amount of caramel colour into a 100- ml volumetric flask with the aid of
0.025 N hydrochloric acid. Dilute to volume with 0.025 N hydrochloric acid
and centrifuge or filter, if solution is cloudy. Take a 20 ml aliquot of the
caramel colour solution, add 200 mg of DEAE Cellulose, mix thoroughly for
several min, centrifuge or filter, and collect the clear supernatant. Determine
the absorbance of the caramel colour solution and the supernatant in a 1-cm
cell at 560 nm, with a suitable spectrophotometer previously standardized
using 0.025 N hydrochloric acid as reference. Calculate the percentage of
Colour Bound by DEAE Cellulose by the formula:
where:
A1 is the absorbance
of the caramel colour solution at 560 nm
A2 is the absorbance
of the supernatant after DEAE Cellulose treatment at 560 nm.
Classification/Colour
bound by phosphoryl cellulose
For the purposes of this
specification colour bound phosphoryl cellulose is defined as the percentage
of decrease in absorbance of a caramel colour solution at 560 nm after
treatment with Phosphoryl Cellulose.
Special reagent:
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Procedure:
Transfer 200-300 mg of caramel
colour into a 100-ml volumetric flask, dilute to volume with 0.025 N
hydrochloric acid, and centrifuge or filter, if solution is cloudy. Take a 40
ml aliquot of the caramel colour solution, add 2.0 g Phosphoryl Cellulose and
mix thoroughly for several min. Centrifuge or filter, and collect the clear
supernatant. Determine the absorbance of the caramel colour solution and the
supernatant in a 1-cm cell at 560 nm, with a suitable spectrophotometer
previously standardized using 0.025 N hydrochloric acid as reference.
Calculate the percentage of Colour Bound by Phosphoryl Cellulose by the
formula:
where:
A1 is the absorbance
of the caramel colour solution at 560 nm
A2 is the absorbance
of the supernatant after Phosphoryl Cellulose treatment at 560 nm.
Classification/
Absorbance ratio
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Procedure:
Transfer 100 mg of caramel colour
into a 100-ml volumetric flask with the aid of water, dilute to volume, mix
and centrifuge if solution is cloudy. Pipet a 5.0 ml portion of the clear
solution into a 100-ml volumetric flask, dilute to volume with water, and
mix. Determine the absorbance of the 0.1% solution in a 1-cm cell at 560 nm
and that of the 1:20 diluted solution at 280 nm with a suitable
spectrophotometer previously standardized using water as reference. (A
suitable spectrophotometer is one equipped with a monochromator to provide a
band width of 2 nm or less and of such quality that the stray-light
characteristic is 0.5% or less.) Calculate the Absorbance Ratio of the
caramel colour by dividing the absorbance units at 280 nm multiplied by 20
(dilution factor) by the absorbance units at 560 nm.
6.2. Purity
Solid
content
The solids content of Caramel
Colour is determined by drying a sample upon a carrier composed of pure
quartz sand that passes a No. 40 but not a No. 60 sieve and has been prepared
by digestion with hydrochloric acid, washed acid-free, dried and ignited. Mix
30.0 g of prepared sand accurately weighed with 1.5-2.0 g Caramel Colour
accurately weighed and dry to constant weight at 60o under reduced pressure
50 mm/Hg (6.7 kPa). Record the final weight of the sand plus caramel. Calculate
the % solids as follows:
% solids = 
where:
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m2 = weight of sand
(g)
m1 = weight of caramel
initially added (g)
Calculation on a solids basis
The contents of Total Nitrogen,
Total sulfur, Ammoniacal nitrogen, sulfur dioxide, 4-MEI and THI are
expressed on a solids basis. The concentration (Ci) of each impurity is
determined on an "as is" basis; the concentration (CS)
on a solid basis is then calculated using the formula:
Ci ´ 100 % solids  Cs =
... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. For the purpose of this
specification, Colour Intensity is defined as the absorbance of a 0.1% (w/v)
solution of Caramel Colour solids in water in a 1 cm cell at 610 nm. Procedure: Transfer 100 mg of Caramel Colour
into a 100 ml volumetric flask, dilute to volume with water, mix and
centrifuge if the solution is cloudy. Determine the absorbance (A610) of the
clear solution in a 1 cm cell at 610 nm with a suitable spectrophotometer
previously standardized using water as a reference. Calculate the Colour
Intensity of the Caramel Colour as follows:
A610 ´ 100 % solids  Colour intensity =
Determine % solids as described
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CS ´
0,1 Colour intensity  Equivalent colour basis =
where: CS = concentration on
a solids basis. This gives content expressed in
terms of a product having a Colour Intensity of 0.1 absorbance units. Total
nitrogen ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Total
sulfur In the largest available
casserole that fits in an electric muffle furnace, place 1-3 g MgO or
equivalent quantity of Mg(NO3)2·6H2O (6.4 -
19.2 g), 1 g powdered sucrose, and 50 ml HNO3. Add 5-10 g caramel colour. Place
same quantities of reagents in another casserole for blank. Evaporate on
steam bath to paste. Place casserole in cold electric muffle (25º) and
gradually heat until all NO2 fumes are driven off. Cool, dissolve and
neutralize with HCl (1+2.5), adding excess of 5 ml. Filter, heat to boiling,
and add 5 ml 10% BaCl2·2H2O solution dropwise.
Evaporate to 100 ml, let stand overnight, filter, wash, ignite, and weigh the
BaSO4. Correct result for BaSO4 obtained in blank and
report as mg S/100 g. Commercial instruments that analyse for total sulfur
such as, the Leco-Combustion/Titration procedure can also be used and are
recommended for sample amounts of about 200 mg. Sulfur
dioxide Apparatus Use a modified Monier-Williams
apparatus (available from 5GA Scientific, Inc., Bloomfield, N.J., USA) for
the determination of sulfurous acid, or construct the apparatus as shown in
the figure. The assembly consists of a 1000-ml three-neck round-bottom
distillation flask having 24/40 standardtaper ground-glass joints. A 30-cm
Allihn condenser is attached in the reflux position to an outer neck of the
flask, and the other end of the condenser is connected with 1/4-inch. Tygon
or silicon tubing (preboiled with 1 in 20 hydrochloric acid solution and
rinsed with water) to the absorption tube assembly (having 35/20 ball joints
or the equivalent). Connect the centre neck of the flask with a 125-ml
cylindrical separator, and attach a piece of tubing to a short U-tube
inserted through a rubber stopper in the neck of the separator. Attach a
curved glass inlet tube, reaching nearly to the bottom of the flask, to the
other outer neck of the flask, and connect the inlet tube to a 250-ml
gas-washing bottle with a piece of the tubing. The gas-washing bottle, in
turn, is connected by tubing to a nitrogen cylinder. 
... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. To each U-tube of the absorption
tube assembly add the following: two pieces of 8-mm glass rod about 25 mm in
length, 10 ml of 3-mm glass beads at the exit side, 10.0 ml of 3% hydrogen
peroxide solution, and 1 drop of methyl red TS. Assemble all pieces of the
apparatus, and check for leaks by blowing gently into the tubing attached to
the neck of the separator. While blowing, close the stopcock of the
separator. Let stand for a few min; if the liquid levels in the U-tubes
equalize, reseal all connections and test again. If the system is airtight
proceed as directed below. Procedure: Disperse about 25 g of the
sample, accurately weighed, in 300 ml of recently boiled and cooled water,
and transfer the slurry to the flask with the aid of water, using a
large-bore funnel. Dilute to about 400 ml with water, and reseal the
separator. Add 90 ml of 4 N hydrochloric acid to the separator, and force the
acid into the flask by blowing gently into the tube in the neck of the
separator. Close the stopcock of the separator. Unclamp the tubing on both sides of the gas-washing bottle, and
start the nitrogen flow at a steady stream of bubbles. Heat the distilling
flask with a heating mantle to cause refluxing in approximately 20 min. When
steady refluxing is reached, apply the line voltage to the mantle and reflux
for 1.75 h. Turn off the water in the condenser, and continue heating until
the inlet joint of the first U-tube shows condensation and slight warming.
Remove the separator and turn off the heat. When the joint at the top of the
condenser cools, remove the connecting assembly and rinse it into the second
U-tube, leaving the crossover tube attached to the exit joint of the first
U-tube but disconnected from the entrance of the second U-tube. Rotate the
crossover tube until the free end almost touches the entrance of the first
U-tube. Add 1 drop of methyl red TS to the first U-tube, and titrate with 0.1
N sodium hydroxide just to a clear yellow colour, mixing with a gentle
rocking motion. After titrating the first Utube, remove the crossover tube,
attach it to the second U-tube exit, and titrate similarly. Record the sum of
the two titers as Vt, in ml. Perform a blank determination,
and record the volume of 0.1 N sodium hydroxide required as Vo. Calculate
the percentage of sulfur dioxide in the sample by the formula 
Where: m is the weight of the sample
taken, in g. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Add 25 ml of 0.1 N sulfuric acid
to a 500-ml receiving flask, and connect it to a distillation apparatus
consisting of a Kjeldahl connecting bulb and a condenser such that the
condenser delivery tube is immersed beneath the surface of the acid solution
in the receiving flask. Transfer about 2 g of caramel colour, accurately
weighed, into an 800-ml long-neck Kjeldahl digestion flask, and to the flask
add 2 g of magnesium oxide (carbonatefree), 200 ml of water, and several
boiling chips. Swirl the digestion flask to mix the contents, and quickly
connect it to the distillation apparatus. Heat the digestion flask to
boiling, and collect about 100 ml of distillate in the receiving flask. Wash
the tip of the delivery tube with a few ml of water, collecting the washings
in the receiving flask, then add 4 or 5 drops of methyl red indicator (500 mg
of methyl red in 100 ml of alcohol), and titrate with 0.1 N sodium hydroxide,
recording the volume, in ml, required as Vt. Conduct a blank
determination, and record the volume, in ml, of 0.1 N sodium hydroxide
required to neutralize as Vo. Calculate the percentage of
ammoniacal nitrogen in the sample by the formula: Ammoniacal nitrogen  Where: m is the weight of the sample
taken, in g. 4-Methylimidazole Note: Information on an improved
method is sought. The following materials and
reagents are required (the reagents should be ACS grade or equivalent where
applicable). ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Pyrex glasswool, 22 x 300 mm
chromatography column with PTFE stopcock (e.g. Kimax 17800); 150 ml
polypropylene beaker (e.g. Nalge 1201); 250 ml round-bottom flask (e.g.,
Pyrex 4320); 75 mm powder funnel; 5 cm spatula; rotary vacuum evaporator; hot
plate; pan for water bath; disposable Pasteur pipets; 5 ml volumetric flask. Reagents Acetone; Celite 545; methylene
chloride; sodium hydroxide; and tetrahydrofuran. Procedure: After thoroughly mixing the
caramel colour sample by shaking or stirring, weigh a 10.00 g aliquot into a
150 ml polypropylene beaker. Polypropylene is considered superior to glass
because of its hydrophobic surface which facilitates quantitative sample
transfer. A 5.0 g portion of 3.0 N NaOH is added and thoroughly mixed to
ensure that the pH of the entire sample exceeds 12. A 20 g portion of Celite
545 is added to the beaker, and the contents are mixed until a semi-dry
mixture is obtained. This normally requires approximately 2 to 3 min. With
samples of unusually high water content, the resultant caramel colour-Celite
545 mixture may be overly wet. In such cases, a 5.00 g aliquot of caramel
colour may be mixed with 2.5 g of 3.0 N NaOH and 15 g of Celite 545 and
carried through the remainder of the analysis. A plug of Pyrex glasswool is
placed in the bottom of a 22 x 300 mm chromatographic column with PTFE
stopcock. The caramel colour-Celite 545 mixture is placed in the column with
the aid of a 75 mm powder funnel. The column contents are settled by
repeatedly allowing the column to fall vertically about 10 cm to a padded
surface. When properly settled, the caramel colour-Celite 545 mixture should
occupy approximately the lower 250 mm of the column. Care should be exercised
at this point to avoid a column bed which is either too loosely or too
tightly packed. Loose packing will allow too rapid elution of the methylene
chloride and possibly incomplete extraction. A too tightly packed column,
e.g., the result of tamping down the column contents, can result in regions
of the bed which are relatively inaccessible to the extraction solvent. This
can also result in incomplete extraction. With the
stopcock open, the column is filled with methylene chloride poured from the
sample beaker. When the solvent reaches the glasswool plug, the stopcock is
closed and the solvent is allowed to stand in contact with the bed for 5 min.
The stopcock is then opened and the column is further eluted with methylene
chloride until 200 ml have been collected in a 250 ml roundbottom flask. A
1.00 ml aliquot of 2 MEI internal standard solution (50.0 mg of 2 MEI/50.0 ml
of methylene chloride) is added to the collected eluate. The 2 MEI is well
separated from the 4 MEI under the GLC conditions employed and has not been
found in caramel colour. The bulk
of the solvent is then removed from the eluate on a rotary vacuum evaporator
operated at 45-50 kPa and with the round-bottom flask maintained at 35o
in a water bath. The extracted residue is transferred quantitatively to a 5
ml volumetric flask with a disposable Pasteur pipet, by rinsing the round-bottom
flask several times with small (ca. 0.75 ml) portions of either
tetrahydrofuran or acetone. Both solvents have been used with equal success.
After mixing the contents thoroughly by several inversions of the flask, the
extract is ready for GLC analysis. The extracts should be analysed as soon as
possible after their preparation, because stability problems have
occasionally been encountered with extracts more than 1 day old. The GLC analysis is carried out
using a gas chromatograph equipped with a hydrogen flame detector. The column
is glass, 1 mm x 6 mm o.d. x 4 mm i.d., filled with 7.5% Carbowax 20M + 2%
K0H on 90/100 mesh Anakrom ABS. The GLC parameters are as follows: carrier,
nitrogen, 50 ml/min; hydrogen, 50 ml/min; oxygen, 80 ml/min; injection port,
200o ; column isothermal, 180o ; detector, 250o
; sample size, 5 µl. All quantitation is done by using the internal standard
technique. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Note: Information on an improved
method is sought. THI is
converted into its 2,4-dinitrophenylhydrazone (THI-DNPH). This derivative is
separated from excess reagent and carbonylic contaminants by HPLC on RP-8,
then determined by its absorbance at 385 nm. Procedure: Caramel colour (200-250 mg) is
weighed accurately, then dissolved in water (3 ml). The solution is
transferred quantitatively to the upper part of a Combination Column. Elution
with water is started, and a total of about 100 ml of water is passed through
the columns. The upper
column is then disconnected. The lower column is eluted with 0.5 N HCl. The
first 10.0 ml of eluate are discarded, then a volume of 35 ml is collected. The solution is concentrated to
dryness at 40o and 15 torr. The syrup residue is dissolved in
carbonyl-free methanol (250 µl) and the 2,4- dinitrophenylhydrazine reagent
(250 µl) is added. The reaction mixture is transferred to a septum-capped
vial and stored for 5 h at room temperature. A volume of 5 µl (but also from
1 to 25 µl) is injected onto a LiChrosorb RP-8 (10 µm) HPLC column. The mobile
phase is MeOH/0.1 M H3PO4 50/50 (v/v). Adjustments in mobile phase
composition may be needed as column characteristics vary, depending upon the
manufacturer. (Use of LiChrosorb RP-8, 10 µm, 250 x 4 mm "Vertex"
column manufactured by Knauer, Bad Homburg, F.R.G. is strongly recommended).
At a mobile phase flow rate of 2 ml/min and column dimensions of 250 x 4.6
mm, THI-DNPH is eluted at about 6.3±0.1 min. It is detected at 385 nm and the
peak height is measured. The amount is calculated from a calibration curve
prepared with THI-DNPH in methanol. Materials: - 2,4,-Dinitrophenylhydrazine
hydrochloride reagent: Commercial 2,4- dinitrophenylhydrazine (5 g) is added
to concentrated hydrochloric acid (10 ml) in a 100-ml Erlenmeyer flask, and
the latter is gently shaken until the free base (red) is converted to the
hydrochloride (yellow). Ethanol (100 ml) is added and the mixture is heated
on a steam bath until all the solid has dissolved. After crystallization at
room temperature, the hydrochloride is filtered off, washed with ether, dried
at room temperature and stored in a desiccator. On storage the hydrochloride
is slowly converted to the free base. The latter can be removed by washing
with dimethoxyethane. The reagent is prepared by mixing 0.5 g of 2,4-dinitrophenylhydrazine
hydrochloride in 15 ml of 5% methanol in dimethoxyethane for 30 min. It
should be stored in the refrigerator and be checked periodically by HPLC. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. - Cation-exchange resin (weak):
Amberlite CG AG 50 I, H+ , (100-200 mesh). (Sediment two or three times prior
to use). - Methanol, carbonyl-free:
Methanol is prepared after Y. Peleg and C.H. Mannheim, J. Agr. Fd. Chem, 18
(1970) 176, by treatment with Girard P reagent. - Dimethoxyethane: If impure,
dimethoxyethane is purified by distillation from 2,4-dinitrophenylhydrazine
in the presence of acid and redistilled from sodium hydroxide. Immediately
prior to use it is passed through a column of neutral aluminium to remove
peroxides. Apparatus: Combination Columns: Similar to
the set-up described in J. Agr. Fd. Chem. 22 (1974) 110. The upper column
(150 x 12.5 mm, filling height max. 9 cm, or 200 x 10 mm, filling height max.
14 cm, with capillary outlet of 1 mm i.d.) is filled with weakly acidic
cation-exchanger; bed height, approx. 50-60, or 80-90 mm, respectively. The
lower column (total length 175 mm, i.d. 10 mm, with capillary outlet and
Teflon stopcock) is filled with strongly acidic cationexchanger to a
bed-height of 60 mm. As a solvent reservoir, a dropping funnel (100 ml) with
Teflon stopcock is used. All parts are connected by standard ground-glass
joints (14.5 mm). HPLC:
With the column specified above and an ultraviolet detector capable of
reading at 385 nm. Calibration: THI-DNPH is
dissolved in absolute, carbonyl-free methanol (about 100 mg/l; calculated
concentration of THI: 47.58 ng/µl). A portion of this solution is diluted
tenfold with methanol (4.7 ng THI/µl). THI-DNPH standard solutions are stable
for at least twenty weeks when stored in the refrigerator. Arsenic ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. - Tested
as directed under JECFA monograph 1-Vol. 4. - Method
II. Lead - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.” ANNEX 20 SPECIFICATIONS AND TESTS FOR SUNSET BRILLIANT BLUE FCF 1. Synonyms ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. INS: 151 ADI = 0 -
1 mg/kg bw. 2. Definition Consists essentially of
tetrasodium 4-acetamido-5-hydroxy-6-[7-sulfonato-4-
(4-sulfonato-phenylazo)-1-naphthylazo]-1,7-naphthalene-disulfonate and
subsidiary colouring matters together with sodium chloride and/or sodium
sulfate as the principal uncoloured components. May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies. Chemical
names Tetrasodium
4-acetamido-5-hydroxy-6-[7-sulfonato-4-(4-sulfonatophenylazo)-
1-naphthylazo]-1,7-naphthalene-disulfonate C.A.S.
number 2519-30-4 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. C28H17N5Na4O14S4 Structural
formula 
Formula
weight 867.69 3. Description Black powder
or granules 4. Functional
uses Colour. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 5.1. Identification Solubility Soluble
in water; sparingly soluble in ethanol Identification
of colouring matters Passes
test. 5.2. Purity Loss on
drying at 135 °C ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Water
insoluble matter Not more
than 0.2%. Lead Not more than 2 mg/kg. Subsidiary
colouring matters Not more
than 4%. Organic
compounds other than colouring matters Not more than 0.8%, sum of
4-Acetamido-5-hydroxy-1,7-naphthalenedisulfonic acid,
4-Amino-5-hydroxy-1,7-naphthalenedisulfonic acid,
8-Amino-2-naphthalenesulfonic acid, Sulfanilic acid,
4,4'-Diazoaminodi(benzenesulfuric acid) Unsulfonated primary
aromatic amines ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Ether
extractable matter Not more
than 0.2%. 5.3. Content Not less
than 80% total colouring matter. 6. Tests 6.1.
Purity Lead - Tested as directed under JECFA
monograph 1-Vol. 4. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Subsidiary
colouring matters - Tested
as directed under JECFA monograph 1-Vol. 4. - Use the
following conditions: Developing
solvent: Chromatogram (i): No. 1. Chromatogram
(ii): No. 4. Height of
ascent of solvent front: (i): approximately 17 cm (ii):
approximately 17 cm Organic
compounds other than colouring matters - Tested as directed under JECFA
monograph 1-Vol. 4. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 6.2.
Method of assay Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following: Weight of
sample: 0.6 - 0.7 g; Buffer:
15 g sodium hydrogen tartrate; Weight
(D) of colouring matters equivalent to 1.00 ml of 0.1 N TiCl3:
10.86 mg. ANNEX 21 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 1. Synonyms CI Food
Brown 3, Chocolate brown HT, CI (1975) No. 20285 INS: 155 ADI = 0 -
1,5 mg/kg bw. 2. Definition Consists
essentially of disodium 4,4'- (2,4-dihydroxy-5-hydroxymethyl-1,3-
phenylene-bisazo) di-1-naphthalene-sulfonate and subsidiary colouring matters
together with NaCl and/or Na2SO4 as the principal
uncoloured components. May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies. Chemical
names Disodium
4,4'-(2,4-dihydroxy-5-hydroxymethyl-1,3-phenylene-bisazo di-1-
naphthalenesulfonate. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 4553-89-3 Chemical
formula C27H18N4Na2O9S2 Structural
formula 
Formula
weight 652.57 3. Description Brown powder
or granules. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Colour. 5. Specifications 5.1. Identification Solubility Soluble
in water; insoluble in ethanol. Identification
of colouring matters Passes
test. 5.2. Purity ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Loss on
drying at 135 °C Not more
than 30% together with chloride and sulfate calculated as sodium salts. Water
insoluble matter Not more
than 0.2%. Lead Not more than 2 mg/kg. Subsidiary
colouring matters Not more than 10%. Organic
compounds other than colouring matters ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Unsulfonated primary
aromatic amines Not more
than 0.01% calculated as aniline. Ether
extractable matter Not more
than 0.2%. 5.3. Content Not less
than 70% total colouring matter. 6. Tests 6.1
Purity ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.” Subsidiary
colouring matters - Tested as directed under JECFA
monograph 1-Vol. 4. - Use the following conditions: Prepare the standard in the
following manner: Dilute 1.0 ml of the 1% dye solution to 100 ml with water
and mix well. Transfer 0.10 ml of this solution to a test tube; add 5.0 ml of
water: acetone (1:1 by vol.) and then 14.9 ml of 0.05 N sodium hydrogen
carbonate solution and shake the tube to ensure mixing. Determine the net
absorbance (AS) of the standard. Developing solvent: No.6. Develop chromatogram for
approximately 14 h. Organic
compounds other than colouring matters ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. - Use liquid chromatography under
the following conditions: HPLC elution gradient: 2 to 100% at 2.0% per min. 6.2.
Method of assay Proceed as directed under Total
Content by Spectrophotometry (in JECFA monograph 1-Vol. 4), using the
following conditions: Solvent:
pH 7 phosphate buffer; Dilution
of solution A: 10 ml to 250 ml. Absorptivity
(a): 40.3 Approximate
wavelength of maximum absorption: 460 nm. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. ANNEX 22 SPECIFICATIONS AND TESTS FOR β-CAROTENE, SYNTHETIC 1. Synonyms CI food
Orange 5; CI (1975): 40800 INS:
160ai ADI = 0 -
5 mg/kg bw. 2. Definition These specifications apply to
predominantly all trans (Z) isomer of b-carotene together with minor
amounts of other carotenoids; diluted and stabilized forms are prepared from b-carotene
meeting these specifications and include solutions or suspensions of b-carotene
in edible fats or oils, emulsions and water dispersible powders; these
preparations may have different cis/trans isomer ratios; the analytical
methods described for the parent colour are not necessarily suitable for the
assay of or determination of impurities in the stabilized forms (appropriate
methods should be available from the manufacturer). Chemical
names ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. C.A.S.
number 7235-40-7 Chemical
formula C40H56 Structural
formula 
Formula
weight 536.88 3. Description ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 4. Functional
uses Colour. 5. Specifications 5.1. Identification Solubility Insoluble
in water; practically insoluble in ethanol; slightly soluble in vegetable
oils; soluble in chloroform. Spectrophotometry Determine the absorbance of the
sample solution C (See Method of Assay) at 455 nm and 483 nm. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Determine the absorbance of the
sample solution C at 455 nm and that of sample Solution B (See Method of
Assay) at 340 nm. The ratio
A455/A340 is not lower than 15. Test for
carotenoid Passes test (described in
“Tests”). Carr-Price
reaction A solution of the sample in
chloroform turns blue on addition of an excess of Carr-Price reagent TS. 5.2. Purity Sulfated
ash ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Subsidiary
colouring matters Carotenoids other than b-carotene:
Not more than 3% of total colouring matters. Lead Not more than 2 mg/kg. 5.3. Content Not less
than 96% total colouring matters, expressed as b-carotene. 6. Tests 6.1.
Identification ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. The colour of a solution of the
sample in acetone disappears after successive additions of a 5% solution of
sodium nitrite and 1 N sulfuric acid. 6.2.
Purity Sulfated
ash - Tested as directed under JECFA
monograph 1-Vol. 4. - Test 2 g of the sample (Method
I) Subsidiary
colouring matters Carotenoids other than b-carotene Dissolve about 80 mg of sample in
100 ml chloroform. Apply 400 µl of this solution as a streak 2 cm from the
bottom of a TLC-plate (Silicagel 0.25 mm). Immediately develop the
chromatogram with a solvent mixture of 95 parts dichloromethane and 5 parts
diethyl ether in a saturated chamber, suitably protected from light, until
the solvent front has moved 15 cm above the initial streak. Remove the plate,
allow the main part of the solvent to evaporate at room temperature and mark
the principal band as well as the bands corresponding to other carotenoids. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Remove the silicagel adsorbent
that contains the combined bands corresponding to the other carotenoids,
transfer it to a glass-stoppered, 50 ml centrifuge tube and add 20.0 ml
chloroform (solution 2). Shake the centrifuge tubes by
mechanical means for 10 min and centrifuge for 5 min. Dilute 10.0 ml of
Solution 1 to 50.0 ml with chloroform (solution 3). Determine, with a
suitable spectrophotometer, the absorbances of Solutions 2 and 3 in 1-cm
cells at the wavelength maximum about 464 nm, using chloroform as blank. Calculation: Calculate the percentage of
carotenoids other than b-carotene (%) = 
where: A2 = absorbance of
Solution 2 A3 = absorbance of
Solution 3 Lead ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.” 6.2.
Method of assay Proceed as directed under Total
Content by Spectrophotometry in JECFA monograph 1-Vol. 4 using the following
conditions: W = 0.08 g V1 = V2
= V3 = 100 ml v1 = v2 = 5
ml A1%1cm =
2500 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. ANNEX 23 SPECIFICATIONS AND TESTS FOR CAROTENES, VEGETABLE 1. Synonyms Natural b-carotene,
carotenes-natural; CI Food Orange 5, mixed carotenes; CI (1975) No. 75130; CI
(1975) No. 40800 (b-Carotene). INS
160aii ADI
"acceptable", provided the level of use does not exceed the level
normally found in vegetables. 2. Definition Carotenes (vegetable) are
obtained by solvent extraction of carrots (Daucus carota), oil of palm
fruit (Elaeis guinensis), sweet potato (Ipomoea batatas) and
other edible plants with subsequent purification. The main colouring
principles are a- and b-carotenes of which b-carotenes account for the major
part. Minor amounts of g- and d-carotenes and other pigments may be present. Besides the colour
pigments, this substance may contain oils, fats and waxes naturally occurring
in the source material. The only solvents used for the extraction are
acetone, methanol, ethanol, propan-2-ol, hexane, carbon dioxide and vegetable
oils. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Class Carotenoid C.A.S.
number 7235-40-7 Chemical
formula C40H56
(-β carotene) Structural
formula 
All-trans-b-carotene ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 536.88 (b-Carotene) 3. Description Red-brown to brown or orange to
dark orange solid or liquid. 4. Functional
uses Colour. 5. Specifications 5.1. Identification Solubility ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Spectrophotometry A
cyclohexane solution of the sample (1 in 200,000) shows maximum absorptions
at 440-457 and 470-486 nm. Colour
reaction Passes test (described in
“Tests”). 5.2. Purity Residual
solvents Not more
than 50 mg/kg, singly or in combination, of acetone, hexane, methanol,
ethanol and propan-2-ol. Lead ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 5.3. Content Content of carotenes (calculated
as b-carotene)
is not less than declared. 6. Tests 6.1.
Identification Colour
reaction A spot of a solution of the
sample in toluene (about 400 µg /ml of ß-carotene) on a filter paper turns
blue 2-3 min after application of a spray or drop of 20% solution of antimony
trichloride solution in toluene. 6.2
Purity ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph 1-Vol. 4, “Instrumental Methods.” 6.3.
Method of assay Proceed as directed under Total
Content by Spectrophotometry in JECFA monograph 1-Vol. 4 using the following
conditions: W =
amount of the sample to obtain adequate absorbance (g) V1
= V2 = V3 = 100 ml v1
= v2 = 5 ml A1%1cm
= 2500 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. ANNEX 24 SPECIFICATIONS AND TESTS FOR CARAMEL EXTRACTS 1. Synonyms Annatto F; Orlean; Terre
orellana; R. Orange; CI (1975) No. 75120 (Natural Orange 4). INS
160bii ADI = 0 -
12 mg/kg bw (for bixin); ADI = 0 -
0.6 mg/kg bw (for norbixin and its salts) The colouring matters bixin and
norbixin derived from annatto extracts (solvent-extracted bixin;
solvent-extracted norbixin; aqueous-processed bixin; alkali-processed
norbixin, acid-precipitated; and alkali-processed norbixin, not
acid-precipitated) are included in the ADIs for bixin and norbixin. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Alkali-processed norbixin
(acid-precipitated) is prepared by removal of the outer coating of the seeds
of the annatto tree (Bixa orellana L) with aqueous alkali. The bixin
is hydrolysed to norbixin in hot alkaline solution and is acidified to
precipitate the norbixin. The precipitate is filtered, dried and milled to
give a granular powder. Alkali-processed norbixin
contains several coloured components; the major colouring principle is
cis-norbixin, a minor colouring principle is trans-norbixin; thermal
degradation products of norbixin may also be present as a result of
processing. Products supplied to the food
industry may be formulated with appropriate carriers of food grade quality. Chemical names cis-Norbixin:
6,6'-Diapo-Ψ,Ψ-carotenedioic acid cis-Norbixin
dipotassium salt: Dipotassium 6,6'-diapo-Ψ,Ψcarotenedioate cis-Norbixin
disodium salt: Disodium 6,6'-diapo-Ψ,Ψ-carotenedioate C.A.S.
number cis-Norbixin:
542-40-5 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. cis-Norbixin
disodium salt: 33261-81-3 Chemical
formula C24H28O4;
C24H26K2O4; C24H26Na2O4 Structural
formula 
Formula
weight 380.5
(acid), 456.7 (dipotassium salt), 424.5 (disodium salt) 3. Description Dark red-brown to red-purple
powder. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Colour. 5. Specifications 5.1. Identification Solubility Soluble
in alkaline water, slightly soluble in ethanol. UV/VIS
absorption The
sample in 0.5% potassium hydroxide solution shows absorbance maxima at about
453 nm and 482 nm. Thin-layer
chromatography ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 5.2. Purity Arsenic Not more than 3 mg/kg. Lead Not more than 2 mg/kg. Mercury Not more
than 1 mg/kg. 5.3. Content ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 6. Tests 6.1.
Identification Thin-layer
chromatography Activate a TLC plate (e.g. LK6D
SILICA GEL 60 A (layer thickness: 250 μm, size: 5 x 20 cm)) for 1 h at 110˚.
Prepare a 5% solution of the sample in 95% ethanol and apply 10 µl to the
plate. Allow to dry and develop using a mixture of n-butanol, methyl ethyl
ketone and 10% aqueous ammonia (3:2:2 by volume) until the solvent front has
ascended about 10 cm. Allow to dry. Bixin and norbixin appear as yellow spots
with Rf values of about 0.50 to 0.45, respectively. Spray with 5%
sodium nitrite solution and then with 0.5 mol/l sulfuric acid and the spots
immediately decolourise. 6.2
Purity Arsenic - Tested as directed under JECFA
monograph 1-Vol. 4. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Lead - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an AAS ICP-AES
technique appropriate to the specified level. The selection of sample size
and method of sample preparation may be based on the principles of the
methods described in JECFA monograph 1-Vol. 4. Mercury - Tested
as directed under JECFA monograph 1-Vol. 4. - Determine
using cold vapour atomic absorption technique. Select sample size appropriate
to the specified level. 6.3.
Method of assay ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Solvent:
0.5 % potassium hydroxide Absorbance
at lmax: about
482 nm. Specific
absorbance A1%1cm = 2870. ANNEX 25 SPECIFICATIONS AND TESTS FOR β-APO-CAROTENAL 1. Synonyms CI food
Orange 6; CI (1975) No. 40820 INS 160e ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 2. Definition Specifications apply to
predominantly all trans (Z) isomer of b-apo-8'-carotenal together with
minor amounts of other carotenoids; diluted and stabilized forms are prepared
from b-apo-8'-carotenal
meeting these specifications and include solutions or suspensions of b-apo-8'-carotenal
in edible fats or oils, emulsions and water dispersible powders; these
preparations may have different cis/trans isomer ratios; the analytical
methods described for the parent colour are not necessarily suitable for the
assay of or determination of impurities in the stabilized forms (appropriate
methods should be available from the manufacturer). Chemical
names b-apo-8’-Carotenal,
8’-apo-b-carotenal C.A.S.
number 1107-26-2 Chemical
formula C30H40O Structural
formula ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Formula
weight 416.65 3. Description Deep violet crystals with
metallic lustre or crystalline powder; sensitive to oxygen and light and
should therefore be kept in a light-resistant container under inert gas. 4. Functional
uses Colour. 5. Specifications 5.1. Identification ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Insoluble
in water, slightly soluble in ethanol, sparingly soluble in vegetable oils,
soluble in chloroform. Spectrophotometry Determine the absorbance of the
sample solution (See Method of Assay) at 461 nm and 488 nm. The ratio A488/A461
is between 0.80 and 0.84. Carotenoid Passes test (described in
“Tests”). Carr-Price
reaction A solution of the sample in chloroform
turns blue on addition of an excess of Carr-Price reagent TS. 5.2. Purity ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Sulfated
ash Not more than 0.1%. Subsidiary
colouring matters Not more than 3% of total
colouring matters. See description under TESTS. Lead Not more than 2 mg/kg. 5.3. Content Not less
than 96% of total colouring matters. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 5.1.
Identification Carotenoid The colour of a solution of the
sample in acetone disappears after successive additions of a 5% solution of
sodium nitrite and 1 N sulfuric acid. 6.2. Purity Sulfated
ash - Tested as directed under JECFA
monograph 1. Vol.4. - Test 2 g of the sample (Method
I) ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Dissolve about 80 mg of sample in
100 ml chloroform. Apply 400 µl of this solution as a streak 2 cm from the
bottom of a TLC-plate (Silicagel 0.25 mm). Pretreat the thin-layer plate by
soaking in a tank with 3% KOH in methanol so that it is completely wetted.
Then dry the plate for 5 min in the air and activate for 1 h at 110o
in an oven. Let cool over CaCl2 and keep in a desiccator over CaCl2.
Immediately after applying the
carotenoid solution to the plate, develop the chromatogram with
n-hexane/chloroform/ethylacetate (70+20+10) in a saturated chamber suitably
protected from light, until the solvent front has moved 10 cm above the
initial streak. Remove the plate, allow the main part of the solvent to
evaporate at room temperature and mark the principal band as well as the
bands corresponding to other carotenoids Remove the silicagel adsorbent
that contains the principal band, transfer it to a glass-stoppered 100 ml
centrifuge tube and add 40.0 ml chloroform (solution 1). Remove the silicagel adsorbent
that contains the combined bands corresponding to the other carotenoids,
transfer it to a glass-stoppered, 50 ml centrifuge tube and add 20.0 ml
chloroform (solution 2). Shake the centrifuge tubes by
mechanical means for 10 min and centrifuge for 5 min. Dilute 10.0 ml of
Solution 1 to 50.0 ml with chloroform (solution 3). Determine, with a
suitable spectrophotometer, the absorbances of Solutions 2 and 3 in 1-cm
cells at the wavelength maximum in chloroform at about 474 nm, using
chloroform as a blank. Calculation: Carotenoids other than b-apo-8'-carotenal: 
where: ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. A3 = absorbance of
Solution 3 Lead - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.” 6.3.
Method of assay Weigh accurately about 80 mg of
the sample and proceed as directed under Total Content by Spectrophotometry
in JECFA monograph 1-Vol. 4 with the following conditions. Absorptivity (a) = 2640 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. ANNEX 26 SPECIFICATIONS AND TESTS FOR β-APO-8'-CAROTENOIC ACID ETHYL ESTER 1. Synonyms CI food
Orange 7; CI (1975) No. 40825 INS 160f ADI = 0 -
5 mg/kg bw. 2. Definition Specifications apply to
predominantly all trans (Z) isomer of ß-apo-8'- carotenoic acid ethyl ester
together with minor amounts of other carotenoids; diluted and stabilized
forms are prepared from ß-apo-8'-carotenoic acid ethyl ester meeting these
specifications and include solutions or suspensions of ßapo-8'-carotenoic
acid ethyl ester in edible fats or oils, emulsions and water dispersible
powders; these preparations may have different cis/trans isomer ratios. The
analytical methods described for the parent colour are not necessarily
suitable for the assay of or determination of impurities in the stabilized
forms (appropriate methods should be available from the manufacturer). ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. b-Apo-8'-carotenoic
acid ethyl ester, ethyl 8'-apo-b-caroten-8'-oate. C.A.S.
number 1109-11-1 Chemical
formula C32H44O2 Structural
formula 
Formula
weight 460.70 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Red to violet-red crystals or
crystalline powder; sensitive to oxygen and light and should therefore be
kept in a light-resistant container under inert gas. 4. Functional
uses Colour. 5. Specifications 5.1. Identification Solubility Insoluble
in water, very slightly soluble in ethanol, slightly soluble in vegetable
oils, soluble in chloroform. Spectrophotometry ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. The ratio A475/A449
is between 0.82 and 0.86. Carotenoid Passes test (described in
“Tests”). Carr-Price
reaction A solution of the sample in
chloroform turns blue on addition of an excess of Carr-Price reagent TS. 5.2. Purity Sulfated
ash Not more than 0.1%. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more than 3% of total
colouring matters. Lead Not more than 2 mg/kg. 5.3. Content Not less
than 96% of total colouring matters. 6. Tests 5.1.
Identification Carotenoid ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 6.2.
Purity Sulfated
ash Tested as directed under JECFA
monograph 1-Vol. 4 - Method I, test 2 g of the sample Subsidiary
colouring matters Carotenoids other than
ß-apo-8'-carotenoic acid ethyl ester: Dissolve about 80 mg of sample in 100
ml chloroform. Apply 400 µl of this solution as a streak 2 cm from the bottom
of a TLC-plate (Silicagel 0.25 mm). Pretreat the thin-layer plate by soaking
in a tank with 3% KOH in methanol so that it is completely wetted. Then dry
the plate for 5 min in the air and activate for 1 h at 110o in an
oven. Let cool over CaCl2 and keep in a desiccator over CaCl2. Immediately after applying the
carotenoid solution to the plate, develop the chromatogram with
n-hexane/chloroform/ethylacetate (70+20+10) in a saturated chamber suitably
protected from light, until the solvent front has moved 10 cm above the
initial streak. Remove the plate, allow the main part of the solvent to
evaporate at room temperature and mark the principal band as well as the
bands corresponding to other carotenoids. Remove the silicagel adsorbent
that contains the principal band, transfer it to a glass-stoppered 100 ml
centrifuge tube and add 40.0 ml chloroform (solution 1). Remove the silicagel adsorbent
that contains the combined bands corresponding to the other carotenoids,
transfer it to a glass-stoppered, 50 ml centrifuge tube and add 20.0 ml
chloroform (solution 2). ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Calculation: Carotenoids other than b-apo-8'-carotenoic
acid ethyl ester (%): 
where: A2 = absorbance of
Solution 2 A3 = absorbance of
Solution 3 Lead - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in JECFA monograph 1-Vol. 4, “Instrumental Methods.” ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Weigh accurately about 80 mg of
the sample and proceed as directed under Total Content by Spectrophotometry
in JECFA monograph 1-Vol. 4 with the following conditions: Absorptivity (a) = 2550 lmax ~ 449
nm. ANNEX 27 SPECIFICATIONS AND TESTS FOR GRAPE SKIN EXTRACT 1. Synonyms
... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. INS 163 (ii) ADI = 0 - 2.5 mg/kg bw. 2. Definition
Obtained by aqueous extraction of
grape skin or marc after the juice has been expressed from it; contains the
common components of grape juice, namely: anthocyanine, tartaric acid,
tannins, sugars, minerals, etc., but not in the same proportions as found in
grape juice. During the extraction process, sulphur dioxide is added and most
of the extracted sugars are fermented to alcohol; the extract is concentrated
by vacuum evaporation during which practically all the alcohol is removed; a
small amount of sulphur dioxide may be present. Chemical
names The principal colouring matters
are anthocyanins, glucosides of anthocyanidins (2-phenylbenzopyrylium salts)
such as peonidin, malvidin, delphinidin, and petunidin. Chemical
formula Peonidin : C16H13O6
X Malvidin : C17H15O7
X ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Petunidin : C16H13O7
X X: acid moiety Structural
formula 
Peonidin : R = OCH3 ;
R’= H Malvidin : R, R’= OCH3
Delphinidin : R, R’= OH Petunidin : R = OCH3 ;
R’= OH X: acid moiety ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Purplish-red liquid, lump, powder
or paste, having a slight characteristic odour. 4. Functional
uses Colour 5. Specifications 5.1.
Identification Solubility
Soluble in water Spectrophotometry ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Colour
reaction Passes test (described in
“Tests”). 5.2.
Purity Sulfur
dioxide Not more
than 0.005% per 1 colour value. Basic
colouring matters Passes test (described in
“Tests”). Other
acidic colouring matters ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Lead Not more than 2 mg/kg 5.3.
Content The colour intensity is not less
than declared. 6. Tests 6.1.
Identification Colour
reaction Add 0.1 g of the sample to 50 ml
of water and shake thoroughly. Filter if necessary. The solution shows red to
purplish-red colour and it turns to blue or dark green on the addition of
sodium hydroxide TS. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Sulfur
dioxide Distil 1 g of the sample with 100
ml of water and 25 ml of phosphoric acid solution (2 in 7) in a distilling
flask with the Wagner tube (Figure 1). In an absorption flask, place 25 ml of
lead acetate solution (1 in 50) previously prepared. Insert the lower end of
condenser into lead acetate solution in the absorption flask. Distil until
the liquid in the absorption flask reaches about 100 ml and rinse the end of
the condenser with a little amount of water. To the distilled solution add 5
ml of hydrochloric acid and 1 ml of starch TS, and titrate with 0.01 N
iodine. Each ml of 0.01 N iodine is
equivalent to 0.3203 mg of SO2  
Figure
1 : Wagner Tube Basic
colouring matters Add 1 g of the sample to 100 ml
sodium hydroxide solution (1 in 100) and shake well. Take 30 ml of this
solution and extract with 15 ml of ether. Extract this ether extract twice
with each 5 ml of dilute acetic acid TS. The acetic acid extract is
colourless. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Add 1 ml of ammonia TS and 10 ml
of water to 1 g of the sample and following the directions Chromatography
place 0.002 ml of the solution on the chromatographic sheet and dry it. Use a
mixture of pyridine and ammonia TS (2:1 by volume) as developing solvent and
stop the development when the solvent front reaches about 15 cm height from
the point where the sample solution
was placed. No spot is observed at the
solvent front after drying under daylight. If any spot is observed, it should
be decolourized when sprayed with a solution of stannous chloride in hydrochloric
acid (2 in 5). Lead Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph 1-Vol. 4, “Instrumental Methods.” 6.3.
Method of assay In the absence of an assay
method, a measurement of colour intensity by the following method may be
used. Prepare approximately 200 ml of
pH 3.0 citric acid - dibasic sodium phosphate buffer solution: Mix 159
volumes of 2.1% citric acid solution and 41 volumes of 0.16% dibasic sodium
phosphate solution, and adjust the pH to 3.0, using the citric acid solution
or Na2HPO4 solution. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Colour value =  m = weight of sample (g) ANNEX 28 SPECIFICATIONS AND TESTS FOR TITANIUM DIOXIDE 1. Synonyms
Titania, CI pigment white 6, CI
(1975) No. 77891 Titanium dioxide INS 171 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 2. Definition
Titanium dioxide is produced by
either the sulfate or the chloride process. Processing conditions determine
the form (anatase or rutile structure) of the final product. In the sulfate process, sulfuric
acid is used to digest ilmenite (FeTiO3) or ilmenite and titanium
slag. After a series of purification steps, the isolated titanium dioxide is
finally washed with water, calcined, and micronized. In the
chloride process, chlorine gas is reacted with a titaniumcontaining mineral
under reducing conditions to form anhydrous titanium tetrachloride, which is
subsequently purified and converted to titanium dioxide either by direct
thermal oxidation or by reaction with steam in the vapour phase. Alternatively,
concentrated hydrochloric acid can be reacted with the titanium-containing
mineral to form a solution of titanium tetrachloride, which is then further
purified and converted to titanium dioxide by hydrolysis. The titanium
dioxide is filtered, washed, and calcined. Commercial titanium dioxide may
be coated with small amounts of alumina and/or silica to improve the
technological properties of the product. C.A.S.
number 13463-67-7 Chemical
formula TiO2 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 79.88 3. Description White to slightly coloured powder 4. Functional
uses Colour 5. Specifications 5.1.
Identification ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Insoluble in water, hydrochloric
acid, dilute sulfuric acid, and organic solvents. Dissolves slowly in
hydrofluoric acid and hot concentrated sulfuric acid. Colour
reaction Passes test (described in
“Tests”). 5.2.
Purity Loss on drying Not more than 0,5%. At 105°, 3h. Loss on ignition Not more than 1.0% on the dried
basis. At 800°. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more than 2%, either singly
or combined Acid-soluble substances Not more than 0.5%; Not more than
1.5% for products containing alumina or silica. Water-soluble matter Not more than 0,5%. Impurities soluble in 0.5 N hydrochloric acid Antimony Not more than 2 mg/kg ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more than 1 mg/kg Cadmium Not more than 1 mg/kg Lead Not more than 10 mg/kg Mercury Not more than 1 mg/kg 5.3.
Method of assay Not less than 99.0% on the dried
basis (on an aluminium oxide and silicon dioxide-free basis) ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 6.1.
Identification Colour reaction Add 5 ml sulfuric acid to 0.5 g
of the sample, heat gently until fumes of sulfuric acid appear, then cool. Cautiously
dilute to about 100 ml with water and filter. To 5 ml of this clear
filtrate, add a few drops of hydrogen
peroxide; an orange-red colour appears immediately. 6.2.
Purity Acid-soluble substances ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Suspend 5 g of the sample in 100
ml 0.5 N hydrochloric acid and on steam bath for 30 min with occasional
stirring. Filter through a Gooch crucible fitted with a glass fibre filter
paper. Wash with three 10-ml portions of 0.5 N hydrochloric acid, evaporate
the combined filtrate and washings to dryness and ignite at a dull red heat
to constant weight. Water-soluble matter Proceed as directed under acid
soluble substances (above), using water in place of 0.5 N hydrochloric acid. Impurities
soluble in 0.5 N hydrochloric acid Antimony, arsenic, cadmium and lead Tested as directed under JECFA
monograph 1-Vol. 4: General tests. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Determine antimony, cadmium, and
lead using an AAS/ICP-AES technique appropriate to the specified level.
Determine arsenic using the ICP-AES/AAS-hydride technique. Alternatively,
determine arsenic using Method II of the Arsenic Limit Test, taking 3 g of
the sample rather than 1 g. The selection of sample size and method of sample
preparation may be based on the principles of the methods described in JECFA
monograph 1-Vol. 4: general tests. Mercury Determine using the cold vapour
atomic absorption technique. Select a sample size appropriate to the
specified level. Tested as directed under JECFA monograph 1-Vol. 4. Aluminium oxide Reagents
and sample solutions 0.01 N
Zinc Sulfate Dissolve 2.9 g of zinc sulfate
(ZnSO4·7H2O) in sufficient water to make 1000 ml. Standardize
the solution as follows: Dissolve 500 mg of high-purity (99.9%) aluminium
wire, accurately weighed, in 20 ml of concentrated hydrochloric acid, heating
gently to effect solution, then transfer the solution into a 1000-ml
volumetric flask, dilute to volume with water, and mix. Transfer a 10 ml
aliquot of this solution into a 500 ml Erlenmeyer flask containing 90 ml of
water and 3 ml of concentrated hydrochloric acid, add 1 drop of methyl orange
TS and 25 ml of 0.02 M disodium ethylenediaminetetraacetate (EDTA) Add,
dropwise, ammonia solution (1 in 5) until the colour is just completely
changed from red to orange-yellow. Then, add: ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. (b): 10
ml of diammonium hydrogen phosphate solution (150 g of diammonium hydrogen
phosphate in 700 ml of water, adjusted to pH 5.5 with a 1 in 2 solution of
hydrochloric acid, then dilute to 1000 ml with water). Boil the
solution for 5 min, cool it quickly to room temperature in a stream of
running water, add 3 drops of xylenol orange TS, and mix. Using the zinc sulfate solution as titrant, titrate the solution
to the first yellow-brown or pink end-point colour that persists for 5-10
sec. (Note: This titration should be performed quickly near the end-point by
adding rapidly 0.2 ml increments of the titrant until the first colour change
occurs; although the colour will fade in 5-10 sec, it is the true end-point.
Failure to observe the first colour change will result in an incorrect
titration. The fading end-point does not occur at the second end-point.) Add 2 g
of sodium fluoride, boil the mixture for 2-5 min, and cool in a stream of
running water. Titrate this solution, using the zinc sulfate solution as
titrant, to the same fugitive yellow-brown or pink end-point as described
above. Calculate
the titre T of zinc sulfate solution by the formula: T = 18,896 m / V where: T is the
mass (mg) of Al2O3 per ml of zinc sulfate solution m is the
mass (g) of aluminium wire V is the
ml of the zinc sulfate solution consumed in the second titration ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. R is the
ratio of the formula weight of aluminium oxide to that of elemental
aluminium. Sample
Solution A Accurately
weigh 1 g of the sample and transfer to a 250-ml high-silica glass Erlenmeyer
flask. Add 10 g of sodium bisulfate (NaHSO4·H2O).
(Note: Do not use more sodium bisulfate than specified, as an excess
concentration of salt will interfere with the EDTA titration later on in the
procedure.) Begin heating the flask at low heat on a hot plate, and then
gradually raise the temperature until full heat is reached. (Caution:
perform this procedure in a well ventilated area. ) When spattering has
stopped and light fumes of SO3 appear, heat in the full flame of a Meeker
burner, with the flask tilted so that the fusion of the sample and sodium
bisulfate is concentrated at one end of the flask. Swirl constantly until the
melt is clear (except for silica content), but guard against prolonged
heating to avoid precipitation of titanium dioxide. Cool, add 25 ml sulfuric
acid solution (1 in 2), and heat until the mass has dissolved and a clear
solution results. Cool, and dilute to 120 ml with water. Introduce a magnetic
stir bar into the flask. Sample
Solution B Prepare
200 ml of an approximately 6.25 M solution of sodium hydroxide. Add 65 ml of this solution to Sample Solution A,
while stirring with the magnetic stirrer; pour the remaining 135 ml of the
alkali solution into a 500-ml volumetric flask. Slowly,
with constant stirring, add the sample mixture to the alkali solution in the
500-ml volumetric flask; dilute to volume with water, and mix. (Note: If the
procedure is delayed at this point for more than 2 hours, store the contents
of the volumetric flask in a polyethylene bottle.) Allow most of the
precipitate to settle (or centrifuge for 5 min), then filter the supernatant
liquid through a very fine filter paper. Label the filtrate Sample Solution
B. Sample
Solution C Transfer
100 ml of the Sample Solution B into a 500-ml Erlenmeyer flask, add 1 drop of
methyl orange TS, acidify with hydrochloric acid solution (1 in 2), and then
add about 3 ml in excess. Add 25 ml of 0.02 M disodium EDTA, and mix. [Note:
If the approximate Al2O3 content is known, calculate
the optimum volume of EDTA solution to be added by the formula: ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Add,
dropwise, ammonia solution (1 in 5) until the colour is just completely
changed from red to orange-yellow. Then add10 ml each of Solutions 1 and 2
(see above) and boil for 5 min. Cool quickly to room temperature in a stream
of running water, add 3 drops of xylenol orange TS, and mix. If the solution
is purple, yellow-brown, or pink, bring the pH to 5.3 - 5.7 by the addition
of acetic acid. At the desired pH, a pink colour indicates that not enough of
the EDTA solution has been added, in which case, discard the solution and
repeat this procedure with another 100 ml of Sample Solution B, using 50 ml,
rather than 25 ml, of 0.02 M disodium EDTA. Procedure Using the
standardized zinc sulfate solution as titrant, titrate Sample Solution C to
the first yellow-brown or pink end-point that persists for 5-10 sec. (Important:
See Note under “0.01 Zinc sulfate”.) This first titration should
require more than 8 ml of titrant, but for more accurate work a titration of
10-15 ml is desirable. Add 2 g
of sodium fluoride to the titration flask, boil the mixture for 2-5 min, and
cool in a stream of running water. Titrate this solution, using the
standardized zinc sulfate solution as titrant, to the same fugitive
yellow-brown or pink end-point as described above. Calculation: Calculate
the percentage of aluminium oxide (Al2O3) in the sample
taken by the formula: % Al2O3 = 100 × (0,005VT)/m where: V is the
number of ml of 0.01 N zinc sulfate consumed in the second titration, ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. m is the
mass (g) of the sample taken, and 0.005 =
500 ml / (1000 mg/g × 100 ml). Silicon dioxide Accurately
weigh 1 g of the sample and transfer to a 250-ml high-silica glass Erlenmeyer
flask. Add 10 g of sodium bisulfate (NaHSO4 · H2O). Heat gently over a Meeker
burner, while swirling the flask, until decomposition and fusion are complete
and the melt is clear, except for the silica content, and then cool.
(Caution: Do not overheat the contents of the flask at the beginning, and
heat cautiously during fusion to avoid spattering.) To the
cooled melt add 25 ml of sulfuric acid solution (1 in 2) and heat carefully
and slowly until the melt is dissolved. Cool, and carefully add 150 ml of
water by pouring very small portions down the sides of the flask, with
frequent swirling to avoid over-heating and spattering. Allow the contents of
the flask to cool, and filter through fine ashless filter paper, using a 60
degree gravity funnel. Rinse out all the silica from the flask onto the
filter paper with sulfuric acid solution (1 in 10). Transfer the filter paper
and its contents into a platinum crucible, dry in an oven at 120°, and heat
the partly covered crucible over a Bunsen burner. To prevent flaming of the
filter paper, first heat the cover from above, and then the crucible from
below. When the
filter paper is consumed, transfer the crucible to a muffle furnace and
ignite at 1000o for 30 min. Cool in a desiccator, and weigh. Add 2
drops of sulfuric acid (1 in 2) and 5 ml of concentrated hydrofluoric acid
(sp.gr. 1.15), and carefully evaporate to dryness, first on a low-heat hot
plate (to remove the HF) and then over a Bunsen burner (to remove the H2SO4).
Take precautions to avoid spattering, especially after removal of the HF. Ignite
at 1000o for 10 min, cool in a desiccator, and weigh again. Record
the difference between the two weights as the content of SiO2 in
the sample. 6.3. Method of assay ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Accurately
weigh about 150 mg of the sample, previously dried at 105o for 3
hours, and transfer into a 500-ml conical flask. Add 5 ml of water and shake
until a homogeneous, milky suspension is obtained. Add 30 ml of sulfuric acid
and 12 g of ammonium sulfate, and mix. Initially heat gently, then heat
strongly until a clear solution is obtained. Cool, then cautiously dilute
with 120 ml of water and 40 ml of hydrochloric acid, and stir. Add 3 g of
aluminium metal, and immediately insert a rubber stopper fitted with a
U-shaped glass tube while immersing the other end of the U-tube into a
saturated solution of sodium bicarbonate contained in a 500-ml wide-mouth
bottle, and generate hydrogen. Allow to stand for a few minutes after the
aluminium metal has dissolved completely to produce a transparent purple
solution. Cool to below 50o in running water, and remove the
rubber stopper carrying the U-tube. Add 3 ml of a saturated potassium
thiocyanate solution as an indicator, and immediately titrate with 0.2 N
ferric ammonium sulfate until a faint brown colour that persists for 30 seconds is obtained. Perform a blank determination and make any
necessary correction. Each ml
of 0.2 N ferric ammonium sulfate is equivalent to 7.990 mg of TiO2. ANNEX 29 SPECIFICATIONS AND TESTS FOR IRON OXIDES 1. Synonyms
Iron oxides; Iron Oxide yellow: CI Pigment
Yellow 42 and 43; CI(1975) No. 77492; INS No. 172(iii) Iron Oxide Red: CI Pigment Red
101 and 102; CI (1975) No. 77491; INS No. 172(ii) ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. ADI = 0 - 0.5 mg/kg bw. 2. Definition
Iron oxides are produced from
ferrous sulfate by heat soaking, removal of water, decomposition, washing,
filtration, drying and grinding. They are produced in either
anhydrous or hydrated forms. Their
range of hues includes yellows, reds, browns and blacks. The food-quality
iron oxides are primarily distinguished from technical grades by their
comparatively low levels of contamination by other metals; this is achieved
by the selection and control of the source of the iron or by the extent of
chemical purification during the manufacturing process. Chemical
names Iron Oxide Yellow: Hydrated
ferric oxide, hydrated iron (III) oxide Iron Oxide Red: Iron sesquioxide,
anhydrous iron (III) oxide Iron Oxide Black: Ferroso ferric
oxide, iron (II,III) oxide C.A.S.
number ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Iron Oxide Red: 1309-37-1 Iron Oxide Black: 1317-61-9 Chemical
formula Iron Oxide Yellow: FeO(OH) .
xH2O Iron Oxide Red: Fe2O3 Iron Oxide Black: FeO · Fe2O3 Formula
weight 88.85
FeO(OH) 159.70 Fe2O3 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 3. Description Yellow, red, brown or black
powder 4. Functional
uses Colour 5. Specifications 5.1.
Identification Solubility
Insoluble in water and organic
solvents; soluble in concentrated mineral acids. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more than 1,0%. 5.2.
Purity Loss on
drying Iron Oxide Red : Not more than
1.0% (105°, 4 h) Arsenic Not more than 3 mg/kg. Cadmium Not more than 1 mg/kg. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more than 10 mg/kg Mercury Not more than 1 mg/kg 5.3.
Content Not less than 60% of iron. 6. Tests 6.1.
Purity Arsenic,
cadmium and lead ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Mercury - Tested as directed under JECFA
monograph 1-Vol. 4. - Determine using an atomic
absorption technique appropriate to the specified level. The selection of
sample size and method of sample preparation may be based on the principles
of the method described in “Instrumental Methods.” 6.2.
Method of assay Weigh accurately about 0.2 g of
the sample, add 10 ml of 5 N hydrochloric acid and heat cautiously to boiling
in a 200-ml conical flask until the sample has dissolved. Allow to cool, add
6 to 7 drops of 30% hydrogen peroxide solution and again heat cautiously to
boiling until all the excess hydrogen peroxide has decomposed (about 2-3
min). Allow to cool, add 30 ml of water and about 2 g of potassium iodide and
allow to stand for 5 min. Add 30 ml of water and titrate with 0.1 N sodium
thiosulfate adding starch TS as the indicator towards the end of the
titration. Each ml of 0.1 N sodium thiosulfate is equivalent to 5.585 mg of
Fe (III). ANNEX 30 ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 1. Synonyms CI food
green 3,CI (1975): 42053 INSL: 143
ADI = 0 -
25 mg/kg bw. 2. Definition Consists essentially of disodium
3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-
sulfonatobenzyl)amino]phenyl](4-hydroxy-2-sulfonatophenyl)methylene]-2,5-cyclohexadien-1-ylidene]ammoniomethyl]benzenesulfonate
and isomers and subsidiary colouring matters together with water, NaCl and/or
Na2SO4 as the principal uncoloured components. May be converted to the
corresponding aluminium lake in which case only the General Specifications
for Aluminium Lakes of Colouring Matters applies. Chemical
names Disodium
3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-sulfonatobenzyl) amino]
phenyl](4-hydroxy-2-sulfonatophenyl)methylene]-2,5-cyclohexadien-1- ylidene]
ammoniomethyl]-benzenesulfonate; ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. C.A.S.
number 2353-43-9 Chemical
formula C37H34N2Na2O10S3 Structural
formula 
Formula
weight 808.86 3. Description ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. 4. Functional
uses Colour. 5. Specifications 5.1. Identification Solubility Soluble
in water; sparingly soluble in ethanol. Identification
of colouring matters Passes
test. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Loss on
drying at 135°C Not more
than 15% together with chloride and sulfate calculated as sodium salts. Water
insoluble matter Not more
than 0.2%. Lead Not more than 2 mg/kg. Chromium Not more
than 50 mg/kg. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more
than 6%. Organic
compounds other than colouring matters Not more
than 0.5%, sum of 2-, 3-, and 4-Formylbenzenesulfonic acids, sodium salts. Not more
than 0.3%, sum of 3- and 4-[N-Ethyl-N-(4-
sulfophenyl)amino]methylbenzenesulfonic acid, disodium salts. Not more
than 0.5% of 2-Formyl-5-hydroxybenzenesulfonic acid, sodium salt. Unsulfonated primary
aromatic amines Not more
than 0.01% calculated as aniline. Leuco
base Not more than 5.0%. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Not more
than 0.4%. 5.3. Content Not less
than 85% total colouring matter. 6. Tests 6.1.
Purity Lead Tested as directed under JECFA
monograph 1-Vol. 4. Determine using an atomic absorption technique
appropriate to the specified level. The selection of sample size and method
of sample preparation may be based on the principles of the method described
in JECFA monograph 1-Vol. 4, “Instrumental Methods.” Subsidiary
colouring matters ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Note: In the calculation
of the subsidiary colour, it is assumed that its absorptivity at its peak is
the same as that of the parent compound at its peak. Standard solutions must
be prepared and their absorptivities measured within 1 h. Spectrophotometric
measurements of the extracted subsidiary colours must be made as promptly as
possible. Organic
compounds other than colouring matters - Tested as directed under JECFA
monograph 1-Vol. 4. - Proceed as directed under
Column Chromatography, using, for example, the following absorptivities: 3-formylbenzenesulfonic acid:
0.495 mg L-1 cm-1 at 246 nm in dilute HCl 3-[(ethyl)(4-sulfophenyl)amino]methylbenzenesulfonic
acid: 0.078 mg L-1 cm-1 at 277 nm in dilute ammonia 2-formyl-5-hydroxybenzenesulfonic
acid: 0.080 mg L-1 cm-1 at 335 nm in dilute ammonia Leuco
base - Tested as directed under JECFA
monograph 1-Vol. 4. ... ... ... Hãy đăng nhập hoặc đăng ký Thành viên Pro tại đây để xem toàn bộ văn bản tiếng Anh. Absorptivity (a) = 0.156 mg L-1
cm-1 at approx. 625 nm. Ratio = 0.971. 6.2.
Method of assay Proceed as directed under Total
Content by Titration with Titanous Chloride (under JECFA monograph 1-Vol. 4),
using the following: Weight of
sample: 1.9 - 2.0 g; Buffer:
15 g sodium hydrogen tartrate; Weight
(D) of colouring matters equivalent to 1.00 ml 0.1 N TiCl3: 40.45
mg.
Quy chuẩn kỹ thuật Quốc gia QCVN 4-10:2010/BYT về phụ gia thực phẩm - phẩm màu do Bộ trưởng Bộ Y tế ban hành
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TP. HCM, ngày 31/05/2021
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