Amendment to the Enforcement Ordinance of the Food Sanitation Law and the 
Standards and Specifications fo r Foods and Food Additives.  
 
The government of Japan will designate Glutamy l-valyl-glycine as an authorized food 
additive. 
 Summary 
Under Article 10 of the Food Sanitation Law (hereinafter referred to as the “Law”), 
food additives shall not be used or marketed  without authorization by the Minister of 
Health, Labour and Welfare (hereinafter referred to as “the Minister”). In addition, when 
specifications or standards are established for f ood additives based on Article 11 of the Law and 
stipulated in the Ministry of Health, Labour a nd Welfare Notification (Ministry of Health and 
Welfare Notification No. 370, 1959), those additiv es shall not be used or marketed unless they 
meet the standards or specifications.  
 
In response to a request from the Minister, the Committee on Food Additives of the 
Food Sanitation Council that is establishe d under the Pharmaceutical Affairs and Food 
Sanitation Council has discussed the adequacy of the designation of Glutamyl-valyl-glycine as a food additive. Th e conclusion of the committee is outlined 
below.  Outline of conclusion  
The Minister, based on Article 10 of the Law, should designate Glutamyl-valyl-glycine, 
as a food additive unlikely to harm human health, and establish standards for use and 
compositional specifications, based on Ar ticle 11 of the Law (see Attachment). 
    Attachment 
Glutamyl -valyl-glycine 
L--Glutamyl -L-valyl-glycine 
グルタミルバリルグリシン  
 
 
 
Standard for use  
Not established.   Compositional specifications  
Substance name  Glutamyl -valyl-glycine 
Molecular formula  C12H21N3O6 
Molecular weight   303.31 
Chemical name [CAS number]  
(2S)-2-amino-4-{(1 S)-1-[(carboxymethyl)carbamoyl]-2-m ethylpropyl}carbamoylbutanoic  
acid [38837-70-6] 
Content Glutamyl -valyl-glycine, when calculated on the dried basis, contains 95.0–
102.0% of  glutamyl -valyl-glycine (C 12H21N3O6). 
Description   Glutamyl -valyl-glycine occurs as a white to light red powder. 
Identification  Determine the infrared absorption spectrum of Glutamyl -valyl-glycine 
as directed in the Potassium Bromide Disk Method under Infrared Spectrophotometry. 
It exhibits the absorption at wavenumbers of about 3321 cm-1, 3282 cm-1, 1712 cm-1, 
1654 cm-1, 1619 cm-1, and 1541 cm-1. 
Purity  
(1) Lead  Not more than 1.0 g/g as Pb. 
Test Solution  Weigh 4.0 g of Glutamyl-valyl-glycine in a platinum, quartz, or 
porcelain crucible or a quartz beaker. Moisten the sample with diluted sulfuric acid (1 in 
4), and heat by increasing the temperature gradually until the sample  is carbonized and 
white fumes are no longer evolved. If necessary , add diluted sulfuric acid (1 in 4) again, 
and heat until the sample is almost carbon ized. Loosely lid the crucible if necessary, 
heat in an electric furnace by increasing the temperature gradually, and incinerate at 
450–600 C. If any carbonized matter still remain s, crush it with a glass rod, moisten 
with 1 ml of diluted sulfuric acid (1 in 4) and 1 ml of nitric acid, and heat until white fumes are no longer evolved. Then, ignite in the electric furnace to complete 
incineration. To the residue, add 10 ml of diluted hydrochloric acid (1 in 4), and 
evaporate on a water bath to dryness. To th e residue, add a little amount of diluted 
nitric acid (1 in 100), and di ssolve it while heating. After co oling, add diluted nitric acid 
(1 in 100) again to make exactly 10 ml.  
Control Solution  Add water to exactly measured 1 ml of Lead Standard Stock 
Solution to make exactly 100 ml. To exactly me asured 4 ml of this solution, add diluted 
nitric acid (1 in 100) to make exactly 10 ml. 
Procedure  Proceed as directed under Method 1 in the Lead Limit Test. 
(2) Arsenic  Not more than 1.0 g/g as As 2O3. 
Test Solution   To 2.0 g of  Glutamyl -valyl-glycine, add 20 ml of water, and heat to 
dissolve. If necessary, use ultrasonic treatment. 
Apparatus   Use Apparatus B. 
Loss on Drying   Not more than 1.0% (105 C, 1 hour). 
Assay   
Test Solution  and Standard Solution   Weigh accurately about 0.05 g each of 
Glutamyl -valyl-glycine and glutamyl -valyl-glycine for assay, add water to each to 
make exactly 50 ml.  Measure exactly 5 ml ea ch of these solutions, add water to make 
exactly 20 ml each. 
Procedure   Analyze 20 l portions of the test solution and the standard solution by 
liquid chromatography using the operating co nditions given below. Measure the peak 
areas (A T and A S) for the test solution and the standard solution, and calculate the 
content by the formula: 
଺ሻ 
ൌ		Dry	basis	weight	 ሺgሻ	of		Glutamylെvalylെglycine	for	assay
	Dry	basis	weight	 ሺgሻ	of		the	sample்
௦ൈ100  
 
Detector: Ultraviolet spectrop hotometer (wavelength 210 nm). 
Column: A stainless steel tube of about 4. 6 mm internal diameter and about 25 cm 
length. 
Column packing material: 5- m octadecylsilanized silica gel for liquid 
chromatography. 
Column temperature: A constant temperature 30–40 C. 
Mobile phase 
A: Dissolve 6.8 g of potassium dihydrogen phosphate in 1000 ml of water, and 
adjust the pH to 3.0 with phosphoric acid. B: Add 600 ml of acetonitrile to 400 ml of Mobile phase A. 
Concentration gradient (A/B): Mainta in 100/0 for 25 minutes, and run a linear 
gradient from 100/0 to 0/100 over 25 minutes. 
Flow rate: 1.0 ml/minute. 
 
Regents and Solutions  
Glutamyl-valyl-glycine for Assay   C 12H21N3O6   A white to light red powder.  
Content   Not less than 99.0% of Glutamyl -valyl-glycine (C 12H21N3O6) when 
calculated on the dried basis.   
Identification  Determine the infrared absorption spectrum of 
Glutamyl -valyl-glycine for Assay as directed in the Potassium Bromide Disk Method 
under Infrared Spectrophotometry. It exhibits the absorption at wavenumbers of 
about 3321 cm-1, 3282 cm-1, 1712 cm-1, 1654 cm-1, 1619 cm-1, and 1541 cm-1. 
Purity Related substances  No t more than 0.50%.  
Prepare a test solution by addi ng water to 25 mg of Glutamyl -valyl-glycine for assay 
to dissolve and to make 25 ml. Prepare a st andard solution as follows: Dilute exactly 
measured 5 ml of the test solution with wa ter to make 50 ml, and dilute again exactly 
measured 5 ml of the resulting solution  with water to make 50 ml. Analyze a 20 μl 
portion each of the test solution and the standard solution by liquid chromatography 
using operating conditions below. Measure the sum of the areas of all peaks, other than the main peak, for the test solution and the main peak area for the standard solution. 
Calculate the amount of the related substances by the formula:  
Amount	ሺ%ሻof	related	substances	
of	the	areas	of	all	peaks,other	than	main	peak, for	the	test 	solution		
݊݋݅ݐݑ݈݋ݏ	݀ݎ݀݊ܽݐݏ	݄݁ݐ	ݎ݋݂	ܽ݁ݎܽ	݇ܽ݁݌	݊݅ܽܯ
 
Operating conditions  Use the conditions given in the Assay for 
Glutamyl -valyl-glycine. 
Loss on drying   Not more than 1.0% (105 C, 1 hour). 
Assay  To about 0.4 g of Glutamyl -valyl-glycine for assay, accurately weighed, add 3 
ml of formic acid to dissolve, and add 50  ml of acetic acid. Titrate with 0.1 mol/L  
perchloric acid. To confirm the endpoint, usually, a potentiometer is used. Perform a 
blank test to make necessary correction. Then, calculate the content on the dried 
basis.  
Each ml of 0.1 mol/L  perchloric acid=30.33 mg of C 12H21N3O6