Printed on: Thu Feb 09 2023, 12:57:50 AM(EST) Status: Not Yet Official on 09-Feb-2023 DocId: GUID-AC406E18-3767-48C7-850B-C29FF498A6DB_5_en-US
Printed by: Dang Van Vu Official Date: 01-Jun-2023 Document Type: DIETARY SUPPLEMENTS @2023 USPC
Do Not Distribute DOI Ref: nad3p DOI: https://doi.org/10.31003/USPNF_M3372_05_01
1
Acceptance criteria: If the Silver manganese paper shows a
Glutathione gray color, it is not more intense than that of the Standard
(NMT 200 ppm).
• ARSENIC á211ñ: NMT 2 ppm
• CHLORIDE AND SULFATE, Chloride á221ñ: Dissolve 0.7 g in
water to make 15 mL. The solution shows no more chloride
than corresponds to 0.20 mL of 0.020 N hydrochloric acid
(NMT 200 ppm).
• CHLORIDE AND SULFATE, Sulfate á221ñ: Dissolve 0.8 g in
water to make 15 mL. The solution shows no more sulfate
than corresponds to 0.25 mL of 0.020 N sulfuric acid (NMT
C10H17N3O6S 307.32 300 ppm).
al
Pentanoic acid, 2-amino-5-[(R)-1-(carboxymethylamino)-3-
mercapto-1-oxopropan-2-ylamino]-5-oxo, (S); Change to read:
N-(N-L-γ-Glutamyl-L-cysteinyl)glycine CAS RN®: 70-18-8.
• ▲IRON á241ñ, Procedures, Procedure 1▲ (CN 1-Jun-2023): NMT
DEFINITION 10 ppm
Glutathione contains NLT 98.0% and NMT 101.0% of • RESIDUE ON IGNITION á281ñ: NMT 0.1%
ci
glutathione (C10H17N3O6S), calculated on the dried basis. • RELATED COMPOUNDS
Mobile phase: 6.8 g/L of potassium dihydrogen phosphate
IDENTIFICATION with 2.02 g/L of sodium 1-heptane sulfonate. Adjust with
• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared phosphoric acid to a pH of 3.0. Mix 970 mL of this solution
Spectroscopy: 197K with 30 mL of methanol.
• B. OPTICAL ROTATION, Specific Rotation á781Sñ
ffi
System suitability solution: 0.1 mg/mL of USP
Sample solution: 40 mg/mL in water l-Phenylalanine RS, 0.5 mg/mL of USP Glutathione RS, and
Acceptance criteria: −15.5° to −17.5°, at 20° 0.5 mg/mL of USP Ascorbic Acid RS in Mobile phase
ASSAY Standard solution: 0.01 mg/mL of USP Glutathione RS in
• PROCEDURE Mobile phase. [NOTE—This solution has a concentration
Sample: 500 mg of Glutathione previously dried equivalent to 2.0% of that of the Sample solution.]
Blank: 50 mL of metaphosphoric acid (1 in 50)
O Sample solution: 50 mg of Glutathione in 100 mL of
Titrimetric system Mobile phase. [NOTE—Allow the solution to stand for 5 min
(See Titrimetry á541ñ.) before use.]
Mode: Direct titration Chromatographic system
Titrant: 0.1 N iodine VS (See Chromatography á621ñ, System Suitability.)
Endpoint detection: Visual Mode: LC
Analysis: Dissolve the Sample in 50 mL of metaphosphoric Detector: UV 210 nm
e
acid (1 in 50) and titrate with the Titrant. Column: 4.6-mm × 15-cm; 5-µm packing L1
Calculate the percentage of glutathione (C10H17N3O6S) in Column temperature: 30°
Flow rate: Adjust so that the retention time of glutathione
the portion of Glutathione taken:
is about 5 min.
B
Result = [(V U − V B) × N × F × 100]/W Injection volume: 10 µL
System suitability
VU = titrant volume of the Sample (mL) Sample: System suitability solution
VB = titrant volume of the Blank (mL) Suitability requirements
Resolution: NLT 5.0 between the ascorbic acid and
N = titrant normality (mEq/mL)
glutathione peaks; and NLT 5.0 between the
To
F = equivalency factor, 307.32 mg/mEq
glutathione and L-phenylalanine peaks
W = weight of the Sample (mg)
Relative standard deviation: NMT 1.5% for replicate
Acceptance criteria: 98.0%–101.0% on the dried basis injections
Analysis
IMPURITIES Samples: Standard solution and Sample solution
• AMMONIUM Calculate the percentage of any impurity in the portion of
Standard solution: 10 µg of ammonium from a diluted Glutathione taken:
ammonium chloride solution
Sample solution: 50 mg of Glutathione Result = (r U/r S) × (C S/C U) × 100
Silver manganese paper: Immerse strips of slow filter paper
(Whatman® filter paper, grade 5 or equivalent) into a rU = peak response of any peak from the Sample
solution containing 8.5 mg/mL of manganese sulfate and solution other than glutathione
8.5 mg/mL of silver nitrate. Maintain for a few min and rS = peak response of the glutathione peak from the
allow to dry over phosphorus pentoxide protected from Standard solution
acid and alkaline vapors. CS = concentration of USP Glutathione RS in the
Analysis: Transfer the Sample solution and the Standard Standard solution (mg/mL)
solution to separate 25-mL jars fitted with caps, and dissolve CU = concentration of Glutathione in the Sample
in 1 mL of water. Add 0.30 g of magnesium oxide. Close solution (mg/mL)
immediately after placing a 5-mm square piece of Silver
manganese paper, wetted with a few drops of water, under Acceptance criteria
the caps. Swirl, avoiding projections of liquid, and allow to Individual impurity: NMT 1.5% for the impurity with the
stand at 40° for 30 min. relative retention time of about 4
Total impurities: NMT 2.0%
https://online.uspnf.com/uspnf/document/1_GUID-AC406E18-3767-48C7-850B-C29FF498A6DB_5_en-US 1/2
Printed on: Thu Feb 09 2023, 12:57:50 AM(EST) Status: Not Yet Official on 09-Feb-2023 DocId: GUID-AC406E18-3767-48C7-850B-C29FF498A6DB_5_en-US
Printed by: Dang Van Vu Official Date: 01-Jun-2023 Document Type: DIETARY SUPPLEMENTS @2023 USPC
Do Not Distribute DOI Ref: nad3p DOI: https://doi.org/10.31003/USPNF_M3372_05_01
2
SPECIFIC TESTS • LOSS ON DRYING á731ñ
• CLARITY AND COLOR OF SOLUTION Analysis: Dry at 105° for 3 h.
Sample solution: 0.1 g/mL in water Acceptance criteria: NMT 0.5%
Analysis: Using identical tubes of colorless, transparent,
neutral glass with a flat base and an internal diameter of 15– ADDITIONAL REQUIREMENTS
25 mm, compare the liquid to be examined with water, the • PACKAGING AND STORAGE: Preserve in tight containers.
depth of the layer being 40 mm. Compare the colors in • USP REFERENCE STANDARDS á11ñ
diffused daylight, viewing vertically against a white USP Ascorbic Acid RS
background. USP Glutathione RS
Acceptance criteria: The solution is clear and colorless. USP l-Phenylalanine RS
al
ci
ffi
O
e
B
To
https://online.uspnf.com/uspnf/document/1_GUID-AC406E18-3767-48C7-850B-C29FF498A6DB_5_en-US 2/2
