Ferrous Glycinate (Processed With Citric Acid) : Synonyms

FERROUS GLYCINATE (processed with CITRIC ACID)

New specifications prepared at the 61st JECFA (2003), published in FNP 52

Add 11 (2003). The compound is suitable for use as a source of iron,
provided that the total intake of iron does not exceed the provisional
maximum tolerable daily intake of 0.8 mg/kg of body weight for iron from all
sources allocated by the 27th meeting.
SYNONYMS Ferrous bisglycinate

DEFINITION Ferrous glycinate is an iron (II) chelate with an iron to glycine ratio of 1:2.
Ferrous glycinate (processed with citric acid) also contains citric acid. It is
manufactured by reaction of reduced iron with glycine in the presence of citric
acid. The resulting product is spray dried without prior removal of the citric
acid. The substance is highly hygroscopic and may contain water in variable
amounts.
Assay Not less than 21.6% and not more than 23.0% of iron (II) on the dried basis
DESCRIPTION A free-flowing, grey green, hygroscopic, fine powder.
FUNCTIONAL USES Nutrient
CHARACTERISTICS
IDENTIFICATION
Solubility (Vol. 4) Freely soluble in water, practically insoluble in acetone and in ethanol
Ferrous ion (vol. 4) Passes test
Infrared spectrum (Vol. 4) The infrared spectrum of a potassium bromide dispersion of the sample dried
at 105° overnight corresponds to the reference infrared spectrum below.
PURITY
Loss on drying (Vol. 4) Not more than 9% (105°, 3 h)
Citric acid Not more than 19.5% on the dried basis

See description under TESTS
Total iron Between 19 and 24% on the dried basis

Ferric iron Not more than 4% on the dried basis

Nitrogen (Vol. 4) Between 11 and 12% on the dried basis
Lead (Vol. 4) Not more than 1 mg/kg

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 on Volume 4, "Instrumental
methods".
TESTS
PURITY TESTS
Citric acid Determine by HPLC using the following conditions:
Column: C18 reversed phase (4.6 x 100 mm)

Detection: UV at 206 nm.
Mobile phase: acetonitrile: 0.1 M diammonium hydrogen phosphate (3:97).
Flow rate: 1.0 ml/min.
Injection: 10 µl.
Run time: 10 min.
Retention time
for citric acid: about 3 min.
0.1 M diammonium hydrogen phosphate: Aqueous solution of 13.20 g/l of

diammonium hydrogenphosphate, (NH4) 2HPO4, pH adjusted to 2.5 with
phosphoric acid TS.
Standard stock solution: Accurately weigh about 200 mg of anhydrous citric
acid, dissolve in 0.1 M diammonium hydrogen phosphate and dilute to 100.0
ml.
Calibration solution: Dilute 10.00 ml of the standard stock solution to 50.0 ml
with 0.1 M diammonium hydrogenphosphate.
Sample solution: Accurately weigh about 200 mg of the dried sample,
dissolve in 0.1 M diammonium hydrogen phosphate and dilute to 100.0 ml.
Procedure
Separately inject equal amounts of the sample solution and the calibration
solution into the chromatograph. Record the chromatograms and measure
the response of citric acid peak. Calculate the content of citric acid in the
sample by the following formula:
A × WC × 50 ×100
% citric acid on the dried basis = S
A C × WS ×10
where,
A S = Peak area of sample solution
A C = Peak area of calibration solution
W S = Weight of dry sample in g
W C = Weight of anhydrous citric acid in g
Total iron Accurately weigh about 0.500 g of thoroughly homogenised sample, and
place the sample into a digestion vessel. Add 5 ml of concentrated nitric acid,
mix the slurry, and cover the vessel with a watch glass or vapour recovery
device. Without bringing to a boil, heat the sample at 95±5° for a period of
30 to 40 min. If brown fumes evolve following heating for the prescribed
time, add 2 ml of concentrated nitric acid repeatedly, with heating for 15 to 20
min, until brown fumes cease to evolve. Continue heating the sample digest
until the volume has been reduced to about 3 ml, ensuring that the bottom of
the vessel is covered with the sample digest at all times. Remove the
heating vessel from the heating source, and allow its contents to cool
thoroughly. Add 2 ml of concentrated hydrochloric acid to the sample digest,
and cover with a watch glass. Replace the vessel on the heating source, and
reflux the sample digest at 95±5° for 15 to 20 min. Before removing the
vessel from the heating source, verify that the evolving vapour is clear. Add 2
ml of water and bring the solution to boiling. Allow to cool to room
temperature, dilute to 50 ml with water, add 3 g of potassium iodide, shake
well, and let stand in the dark for 5 min. Titrate any liberated iodine with 0.1
N sodium thiosulfate, using starch TS as the indicator; 1 ml of 0.1 N sodium
thiosulfate is equivalent to 5.585 mg of total iron.
Ferric iron Accurately weigh about 5 g of the sample into a 250 ml glass-stoppered
flask, and dissolve in a mixture of 100 ml of water and 10 ml of hydrochloric
acid. To the solution, add 3 g of potassium iodide, shake well, and let stand
in the dark for 5 min. Titrate any liberated iodine with 0.1 N sodium
thiosulfate, using starch TS as the indicator; 1 ml of 0.1 N sodium thiosulfate
is equivalent to 5.585 mg of ferric iron.
METHOD OF Dissolve about 1 g, accurately weighed, of the dried sample in a mixture of

ASSAY 150 ml of water and 10 ml of sulfuric acid TS in a 300-ml flask. Add 1 drop of
orthophenanthroline TS, and immediately titrate with 0.1 N ceric sulfate.
Perform a blank determination in an identical manner, and perform any
necessary correction; 1 ml of 0.1 N ceric sulfate is equivalent to 5.585 mg of
ferrous ion.
FTIR spectrum
The absorptions around 2250 cm-1 are due to CO2 absorption

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FERROUS GLYCINATE (processed with CITRIC ACID)

New specifications prepared at the 61st JECFA (2003), published in FNP 52

SYNONYMS Ferrous bisglycinate

DESCRIPTION A free-flowing, grey green, hygroscopic, fine powder.

FUNCTIONAL USES Nutrient

Ferrous ion (vol. 4) Passes test

Loss on drying (Vol. 4) Not more than 9% (105°, 3 h)

Citric acid Not more than 19.5% on the dried basis

Total iron Between 19 and 24% on the dried basis

Ferric iron Not more than 4% on the dried basis

Nitrogen (Vol. 4) Between 11 and 12% on the dried basis

Lead (Vol. 4) Not more than 1 mg/kg

Citric acid Determine by HPLC using the following conditions:

Column: C18 reversed phase (4.6 x 100 mm)

0.1 M diammonium hydrogen phosphate: Aqueous solution of 13.20 g/l of

METHOD OF Dissolve about 1 g, accurately weighed, of the dried sample in a mixture of

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