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GB/T 13089-2020 (GB/T13089-2020)

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GB/T 13089-2020: PDF in English (GBT 13089-2020)
GB/T 13089-2020
ICS 65.120
B 46
Replacing GB/T 13089-1991
Method for Determination of Oxazolidinethione in
Issued by: State Administration for Market Regulation;
Standardization Administration of PRC.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative References ... 4
3 Principle ... 4
4 Reagents and Materials ... 5
5 Apparatus ... 6
6 Samples ... 6
7 Test Procedures ... 7
8 Test Data Processing ... 7
9 Precision ... 8
This Standard was drafted as per the rules specified in GB/T 1.1-2009.
This Standard replaced GB/T 13089-1991 Method for Determination of
Oxazolidinethione in Feeds.
Compared with GB/T 13089-1991, this Standard has the major differences in technical
contents as follows besides the editorial modifications:
--- Modify the scope of application of the method and increase the limit of
quantification (see Clause 1 of this Edition; Clause 1 of 1991 Edition);
--- Modify the principle of the method (see Clause 3 of this Edition; Clause 2 of 1991
--- Modify the preparation method of white myrosinase (see 4.10 of this Edition; 3.4
of 1991 Edition);
---Modify the dosage and test procedures of white myrosinase (see Clause 7 of this
Edition; Clause 6 of 1991 Edition);
--- Modify test data processing (see Clause 8 of this Edition; Clause 7 of 1991
--- Modify the precision (see Clause 9 of this Edition; Clause 7 of 1991 Edition).
Please note some contents of this document may involve patents. The issuing agency
of this document shall not assume the responsibility to identify these patents.
This Standard was proposed by and under the jurisdiction of National Technical
Committee for Standardization of Feed Industry (SAC/TC 76).
Drafting organizations of this Standard: Sichuan Will Test Technology Co., Ltd.;
Institute of Animal Sciences of Chinese Academy of Agricultural Sciences; Public
Monitoring Center for Agro-Product of Guangdong Academy of Agricultural Sciences;
Tongwei Co., Ltd.; and China Feed Industry Association.
Chief drafting staffs of this Standard: Zhang Fengping, Tong Jianming, Wang Weili, Lei
Baoliang, Yang Fashu, Wang Liwen, Song Tao, and Lu Jiawen.
The historical edition replaced by this Standard is as follows:
--- GB/T 13089-1991.
Method for Determination of Oxazolidinethione
in Feeds
1 Scope
This Standard specifies the UV spectrophotometric method for the determination of
oxazolidinthione in feeds.
This Standard is applicable to the determination of oxazolidinthione in rapeseed and
its processed products, as well as compound feeds, concentrated feeds and
concentrate supplements containing rapeseed and its processed products.
The limit of quantification of this Standard method is 150mg/kg.
2 Normative References
The following documents are essential to the application of this document. For the
dated documents, only the versions with the dates indicated are applicable to this
document; for the undated documents, only the latest version (including all the
amendments) is applicable to this document.
GB/T 5520 Inspection of Grain and Oils - Germination Test of Seeds
GB/T 6682 Water for Analytical Laboratory Use - Specification and Test Methods
GB/T 20195 Animal Feeding Stuffs-Preparation of Test Samples
3 Principle
The glucosinolate in the specimen is hydrolyzed into isothiocyanate under the action
of white myrosinase in the buffer solution with pH 7.0. After extraction with
dichloromethane, the isothiocyanate with hydroxyl reacts with ethanol. The
oxazolidinthione is cyclized and formed, which is determined by ultraviolet
4 Reagents and Materials
Unless otherwise specified, only use analytically-pure reagents.
4.1 Water: Class-II water specified in GB/T 6682.
4.2 Petroleum ether (boiling range 30°C ~60°C).
4.3 Dichloromethane.
4.4 Absolute ethanol.
4.5 Citric acid solution (0.1mol/L): accurately take 4.20g of citric acid (C6H8O7·H2O)
and dissolve and make constant volume by water to 200mL; and mix evenly. Prepare
for current use.
4.6 Disodium hydrogen phosphate (0.2mol/L): accurately take 28.39g of anhydrous
disodium hydrogen phosphate (Na2HPO4) and dissolve and make constant volume by
water to 1L; and mix evenly. Prepare for current use.
4.7 Hydrochloric acid (0.01mol/L): accurately pipette 0.9mL of concentrated
hydrochloric acid; dilute and make constant volume by water to 1L; and mix evenly.
4.8 Sodium hydroxide solution (0.01mol/L): accurately take 0.40g of sodium hydroxide
and dissolve and make constant volume by water to 1L; and mix evenly.
4.9 Buffer solution with pH 7.0: pipette 176.5 mL of citric acid solution (4.5) in a 1000
mL volumetric flask; dilute and make constant volume by disodium hydrogen
phosphate solution (4.6) to 1000 mL; mix evenly. Use hydrochloric acid solution (4.7)
or sodium hydroxide solution (4.8) to adjust the pH to 7.0. Prepare for current use.
4.10 White myrosinase: take 50g of white mustard (Sinapis alba L.) seeds (make the
germination rate test in accordance with GB/T 5520; the germination rate must be
greater than 85% within 72h; and the storage period shall not exceed 2 years); after
crushing, place them in a 500mL beaker; add 100mL of petroleum ether (4.2); stir for
2min; let stand; discard the supernatant liquid. Repeat degreasing for 10 times, make
the fat content less than 2%. Put the solvent in a fume hood to evaporate. And then
crush it again, 80% passed through the 0.28mm test sieve; put in an enclosed
container and store at -18°C below. The validity period is 6 months. In order to ensure
the activity of white myrosinase, the environment temperature shall be kept at 30°C
below during the preparation process. Pay attention to a small amount and multiple
times during the two crushing to prevent the crusher from overheating.
(Above excerpt was released on 2021-01-24, modified on 2021-06-07, translated/reviewed by: Wayne Zheng et al.)
Source: https://www.chinesestandard.net/PDF.aspx/GBT13089-2020