GB 5009.124-2016_English: PDF (GB5009.124-2016)
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| GB 5009.124-2016 | English | 85 |
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National Food Safety Standard -- Determination of Amino Acid in Foods
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GB 5009.124-2016
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| GB/T 5009.124-2003 | English | 70 |
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Determination of amino acids in foods
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GB/T 5009.124-2003
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| Standard ID | GB 5009.124-2016 (GB5009.124-2016) | | Description (Translated English) | National Food Safety Standard -- Determination of Amino Acid in Foods | | Sector / Industry | National Standard | | Classification of Chinese Standard | X09 | | Word Count Estimation | 8,885 | | Date of Issue | 2016-12-23 | | Date of Implementation | 2017-06-23 | | Older Standard (superseded by this standard) | GB/T 5009.124-2003 | | Regulation (derived from) | National Health and Family Planning Commission Notice No.17 of 2016 | | Standard ID | GB/T 5009.124-2003 (GB/T5009.124-2003) | | Description (Translated English) | Determination of amino acids in foods | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C53 | | Classification of International Standard | 67.040 | | Word Count Estimation | 5,562 | | Date of Issue | 2003-08-11 | | Date of Implementation | 2004-01-01 | | Older Standard (superseded by this standard) | GB/T 14965-1994 | | Drafting Organization | Chinese Academy of Preventive Medicine, Institute of Nutrition and Food Hygiene | | Administrative Organization | People's Republic of China Ministry of Health | | Proposing organization | Ministry of Health of the People Republic of China | | Issuing agency(ies) | The People Republic of China Ministry of Health, China National Standardization Management Committee | | Summary | This standard specifies: Determination of amino acids in foods amino acid automatic analyzer methods. This standard applies to: foods aspartic acid, threonine, serine, glutamic Yan, proline, glycine, alanine, stretch histidine. Methionine, isoleucine, nitric acid, leucine, tyrosine, phenylalanine, histidine, lysine and arginine sixteen kinds of amino acids determination. The minimum detection limit of 10pmol This standard does not apply to low protein content Fruits, vegetables, drinks and starchy foods amino acid determination. |
GB 5009.124-2016
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard
– Determination of Amino Acid in Foods
ISSUED ON. DECEMBER 23, 2016
IMPLEMENTED ON. JUNE 23, 2017
Issued by. National Health and Family Planning Commission of PRC;
China Food and Drug Administration.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Principle ... 4
3 Reagents and Materials ... 4
4 Instrument and Equipment ... 5
5 Analytical Procedures ... 6
6 Expression of Analytical Results ... 8
7 Precision ... 10
8 Others ... 10
Appendix A Chromatogram ... 12
Foreword
This Standard replaces GB/T 5009.124-2003 Determination of Amino Acid in Foods.
Compared with GB/T 5009.124-2003, this Standard has the major changes as follows.
--- Modify the standard name into “National Food Safety Standard – Determination
of Amino Acid in Foods”;
--- Expand the applicable scope;
--- Add the limit of detection and quantitative limit of the method;
--- Modify the result calculation formula.
National Food Safety Standard
– Determination of Amino Acid in Foods
1 Scope
This Standard specifies using the amino acid analyzer (ninhydrin post-column
derivatization ion exchange chromatography) to determine the amino acid in foods.
This Standard is applicable to determine the amino acid hydrolyzed by acid in foods,
there are totally 16 kinds of amino acids such as aspartic acid, threonine, serine,
glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine,
phenylalanine, histidine, lysine, and arginine.
2 Principle
The protein in food is hydrolyzed by hydrochloric acid into free amino acid; after being
separated by ion exchange column, it occurs color reaction with ninhydrin solution;
then determine the amino acid content through the visible light spectrophotometer.
3 Reagents and Materials
Unless otherwise specified, the reagents used in this method are analytically pure;
while water is Class-I water stipulated in GB/T 6682.
3.1 Reagents
3.1.1 Hydrochloric acid (HCl). concentration≥36%, guarantee reagent.
3.1.2 Phenol (C6H5OH).
3.1.3 Nitrogen. with purity of 99.9%.
3.1.4 Sodium citrate (Na3C6H5O7•2H2O). guarantee reagent.
3.1.5 Sodium hydroxide (NaOH). guarantee reagent.
3.2 Reagents preparation
3.2.1 Hydrochloric acid solution (6mol/L). take 500mL of hydrochloric acid, dilute with
water to 1000mL, mix evenly.
3.2.2 Refrigerant. mix the commercial salt and ice by the mass ratio of 1.3.
Place the hydrolysis tube into the refrigerant, frozen for 3min~5min; connect to the
suction tube of vacuum pump; pump vacuum (close to 0Pa); then inject nitrogen;
repeatedly pump vacuum, inject nitrogen, after 3 times, seal the cap or tighten the
screw cap under nitrogen injecting state.
Place the sealed hydrolysis tube into 110°C±1°C electric blower thermostat or
hydrolysis furnace; hydrolyze for 22h, then take out; cooling off to the room
temperature.
Open the hydrolysis tube, filter the hydrolysate into 50mL volumetric flask; use small
amount of water to wash the hydrolysis tube for several times; transfer the washing
liquid into the same 50mL volumetric flask; finally use water to make constant volume
to the scale; shake and mix evenly.
Accurately pipette 1.0mL of filtrate into 15mL or 25mL test tube; use the test tube
concentrator or parallel evaporator to reduce pressure to dry at the 40°C~50°C heating
temperature environment; after drying, the residuals shall use 1mL~2mL of water to
dissolve; then reduce pressure to dry; finally evaporate to dryness.
Add 1.0mL~2.0mL of sodium citrate buffer solution with pH 2.2 into the test tube after
drying to dissolve; after shaking and mixing evenly; absorb the solution to get through
the 0.22µm filter film; transfer into the instrument sample-injecting bottle; it is the
sample assay solution for the measurement of the instrument.
5.4 Determination
5.4.1 Instrument conditions
Inject the mixed amino acid standard working solution into the automatic amino acid
analyzer; refer to the test protocol and instrument manual of the amino acid analyzer
stipulated in JJG1064-2011; appropriately adjust the instrument operation procedures,
parameters, and reagent ratio of buffer solution; confirm the instrument operating
conditions.
5.4.2 Chromatographic reference conditions
a) chromatographic column. sulfonic acid cation resin;
b) detection wavelength. 570nm and 440nm.
5.4.3 Determination of specimen
Inject the same volume of amino acid standard working solution and sample assay
solution into the amino acid analyzer; calculate the concentration of amino acid in the
sample assay solution through the peak area by external standard method.
......
GB/T 5009.124-2003
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 67.040
C 53
Replacing GB/T 14965-1994
Determination of Amino Acids in Foods
ISSUED ON. AUGUST 11, 2003
IMPLEMENTED ON. JANUARY 1, 2004
Issued by. Ministry of Health of the PRC;
Standardization Administration of China.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Principle ... 4
3 Reagents ... 4
4 Apparatus ... 5
5 Specimen Treatment ... 6
6 Analytical Procedures ... 6
7 Determination ... 6
8 Result Calculation ... 7
9 Precision ... 7
10 Standard Map ... 8
Foreword
This Standard replaces GB/T 14965-1994 Method for Determination of Amino Acids in
Foods.
Compared with GB/T 14965-1994, this Standard mainly has the following changes.
--- Modify the Chinese name of the standard, which was changed into Determination
of Amino Acids in Foods.
--- Modify the structure of the original standard as per GB/T 20001.4-2001 Rules for
Drafting Standards - Part 4. Methods of Chemical Analysis.
This Standard was proposed by and shall be under the jurisdiction of the Ministry of
Health of the PRC.
Drafting organizations of this Standard. Institute of Nutrition and Food Hygiene,
Chinese Academy of Preventive Medicine.
Chief drafting staffs of this Standard. Jia Jianbin, and Zhao Xihe.
The original standard was initially published in 1994; and this is the first-time
amendment.
Determination of Amino Acids in Foods
1 Scope
This Standard specifies the method of using the automatic amino acid analyzer to
determine the amino acids in foods.
This Standard is applicable to the determination of 16 kinds of amino acids in foods,
such as aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine,
methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, arginine, and
etc.. The minimum detection limit is 10 pmol.
This Standard is not applicable to the determination of amino acids in low-protein-
content fruits, vegetables, beverages, and starchy foods.
2 Principle
Protein in food is decomposed by hydrochloric acid water into the free amino acids;
after being separated by ion exchange column of amino acid analyzer, it occurs color
reaction with ninhydrin solution; then the amino acid content is determined by
spectrophotometer colorimetric method.
3 Reagents
3.1 Concentrated hydrochloric acid. guaranteed reagent.
3.2 6 mol/L hydrochloric acid. concentrated hydrochloric acid is mixed with water by
1+1.
3.3 Phenol. requires re-distillation.
3.4 (0.0025 mol/L) mixed amino acid standard solution (sold by apparatus
manufacturer).
3.5 Buffer solution
3.5.1 Sodium citrate buffer solution with pH2.2. weigh 19.6g of sodium citrate
(Na3C6H5O7•2H2O) and 16.5 mL of concentrated hydrochloric acid; add water and
dilute to 1000 mL; adjust the pH to be 2.2 by concentrated hydrochloric acid or 500 g/L
5 Specimen Treatment
After the specimen collection, use the homogenizer to make it homogenate (or try to
crush the specimen); freeze and preserve in the low-temperature refrigerator; thaw the
specimen before analyzing.
6 Analytical Procedures
6.1 Weighing specimen
Accurately weigh certain amount of specimen with good uniformity, for instance milk
powder, and etc.; accurate to 0.0001g, (specimen protein content shall be within the
range of 10mg~20mg); for the specimen with poor uniformity, for instance the fresh
meat, and etc.; in order to reduce the error, the specimen weighing amount can be
increased, dilute the specimen before measurement. Place the weighed specimen into
the hydrolysis tube.
6.2 Hydrolysis
Add 10 mL ~ 15 mL of 6 mol/L hydrolysis acid (the amount depends on the protein
content of the specimen) into the hydrolysis tube; the specimen with high water content
(for instance, milk) can be added with equivalent volume of concentrated hydrochloric
acid; add 3~4 drops of freshly distilled phenol; place the hydrolysis tube into the
refrigerant to freeze for 3 min ~5 min; then connect to the suction pipe of the vacuum
pump, pump vacuum (close to 0 Pa); inject the high-purity nitrogen, then pump vacuum
again and inject nitrogen; after repeating for three times, seal the hydrolysis tube under
the nitrogen injecting state, or tighten the screw cap, and place the sealed hydrolysis
tube within the 110°C±1°C constant temperature drying oven, hydrolyze for 22h, take
out for cooling.
Open the hydrolysis tube, after filtering the hydrolysate, rinse the hydrolysis tube with
deionized water for several times; transfer all hydrolysate into 50 mL volumetric flask,
and use deionized water to make volume. Take 1 mL of filtrate into 5 mL volumetric
flask; use vacuum dryer to dry at 40°C~50°C; the residue is dissolved by 1 mL ~ 2 mL
of water, then dry again; repeat for twice; finally evaporate to dryness; dissolve by 1
mL of buffer solution with pH2.2; then prepare for the apparatus determination.
7 Determination
Accurately take 0.200 mL of mixed amino acid standard solution; dilute it to 5 mL by
buffer solution with pH2.2; the concentration of such standard diluent is 5.00 nmol/50
µL; which is used for amino acid standard of on-machine determination; use the
standard method beyond the automatic amino acid analyzer to determine the content
......
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