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GB/T 13086-2020 (GBT 13086-2020)

GB/T 13086-2020_English: PDF (GBT13086-2020)
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BASIC DATA
Standard ID GB/T 13086-2020 (GB/T13086-2020)
Description (Translated English) Method for determination of free gossypol in feeds
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B46
Classification of International Standard 65.120
Word Count Estimation 6,628
Date of Issue 2020-11-19
Date of Implementation 2021-06-01
Older Standard (superseded by this standard) GB/T 13086-1991
Drafting Organization Jiangxi Veterinary Drug and Feed Supervision Institute, China Feed Industry Association
Administrative Organization National Gold Standardization Technical Committee (SAC/TC 379)
Regulation (derived from) National Standard Announcement No. 26 of 2020
Proposing organization National Feed Industry Standardization Technical Committee (SAC/TC 76)
Issuing agency(ies) State Administration for Market Regulation, National Standardization Administration

GB/T 13086-2020
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 65.120
B 46
Replacing GB/T 13086-1991
Method of Determination of Free Gossypol in Feeds
(ISO 6866:1985, Animal Feeding Stuffs
– Determination of Free and Total Gossypol, MOD)
ISSUED ON: NOVEMBER 19, 2020
IMPLEMENTED ON: JUNE 01, 2021
Issued by: State Administration for Market Regulation;
Standardization Administration of PRC.
Table of Contents
Foreword ... 3
1 Scope ... 6
2 Normative References ... 6
3 Principle ... 6
4 Reagents and Materials ... 6
5 Apparatus ... 7
6 Samples ... 7
7 Test Procedures ... 8
8 Test Data Processing ... 9
9 Precision ... 10
Appendix A (Informative) Structural Changes between this Standard and ISO
6866:1985 ... 11
Foreword
This Standard was drafted as per the rules specified in GB/T 1.1-2009.
This Standard replaced GB/T 13086-1991 Method for Determination of Free Gossypol
in Feeds.
Compared with GB/T 13086-1991, this Standard has the major technical changes as
follows besides the editorial modifications:
--- Modify the scope of application (see Clause 1 of this Edition; Clause 1 of 1991
Edition);
--- Increase the limit of quantification (see Clause 1 of this Edition);
--- Delete reagents in the commercially available form that were only used for
preparation (see Clause 3 of 1991 Edition);
--- Increase the validity period of aniline and solvent A (see 4.2 of this Edition);
--- Modify the constant temperature water bath in the apparatus into a water bath
(see Clause 5 of this Edition; Clause 5 of 1991 Edition);
---Add preparation method for the cottonseed sample (see Clause 6 of this Edition);
--- Modify the sample-weighing amount (see 7.1 of this Edition; 6.1 of 1991 Edition);
--- Add the calculation formula for correcting absorbance (see 8.1 of this Edition);
--- Modify the calculation formula of free gossypol content in the sample (see 8.2 of
this Edition; 7.1 of 1991 Edition);
--- Add the calculation formula for the results of free gossypol in cottonseed (see
8.2 of this Edition);
--- Modify the significant figures of measurement results (see Clause 8 of this Edition;
7.2 of 1991 Edition);
--- Modify the precision provisions (see Clause 9 of this Edition; 7.3 of 1991 Edition);
--- Add the informative Appendix A (see Appendix A of this Edition).
This Standard uses the redrafting method to modify and adopt the international
standard ISO 6866:1985 Animal Feed Stuffs - Determination of Free and Total
Gossypol.
Compared with ISO 6866:1985, this Standard has major adjustments in structure.
Appendix A lists the clause number comparison between this Standard and ISO
6866:1985.
The technical differences and causes between this Standard and ISO 6866:1985 are
as follows:
--- Regarding the normative references, this Standard adjusts the technical
differences to adapt to China's technical conditions. The adjusted conditions and
technologies are reflected in Clause 2 Normative Reference. The specific
adjustments are as follows:
 Delete the reference document ISO 6497 in ISO 6866:1985;
 Replace ISO 6498 by GB/T 20195 which is equivalent to adopting
international standard;
 Add reference of GB/T 6682.
--- The measurement wavelength of the absorbance value is changed from "the
wavelength of maximum absorption (435nm~445nm)" to "the wavelength of
440nm" (see Clause 3, 7.3 of this Edition; 8.3.1.8 of ISO 6866:1985);
--- Delete the content related to the determination of total gossypol (see 8.3.2 of
ISO 6866:1985);
--- Add the calculation formula of corrected absorbance [see Formula (1) of this
Edition];
--- Delete the related content of total gossypol calculation (see 9.1 of ISO
6866:1985);
--- Modify the calculation formula of free gossypol content in the sample [see
Formula (2) of this Edition; 9.1 of ISO 6866:1985];
--- Add the calculation formula for the results of free gossypol in cottonseed [see
Formula (3) of this Edition];
--- Delete the content of the test report (see Clause 10 of ISO 6866:1985).
This Standard also makes the following editorial modifications:
--- Modify the name of the standard; and delete the determination of total gossypol,
which is suitable for the purpose of sampling in China;
Please note that certain contents of this document may involve patents. The issuing
agency of this document is not responsible for identifying these patents.
Method of Determination of Free Gossypol in Feeds
1 Scope
This Standard specifies the spectrophotometric method for the determination of free
gossypol in feeds.
This Standard is applicable to the determination of free gossypol in compound feeds,
concentrated feeds, concentrate supplements, cottonseed, cottonseed meal, and
other feed materials (except cottonseed oil) containing cottonseed and cottonseed
meal.
The limit of quantification of this Standard is 20mg/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 6682 Water for Analytical Laboratory Use - Specification and Test Methods
(GB/T 6682-2008, ISO 3696:1987, MOD)
GB/T 20195 Animal Feeding Stuffs - Preparation of Test Samples (GB/T 20195-
2006, ISO 6498:1998, IDT)
3 Principle
Extract the free gossypol in the sample by a mixture of isopropanol-n-hexane
containing 3-Aminopropanol; and use the aniline to convert the gossypol into aniline
gossypol with aniline. Measure the absorbance value at a wavelength of 440nm.
4 Reagents and Materials
Unless otherwise specified, only the analytically-pure reagents shall be used.
4.1 Water: Class-II specified in GB/T 6682.
7 Test Procedures
7.1 Extraction of sample solution
7.1.1 Sample weighing
Do two tests in parallel. The sample weights for different tests are as follows:
--- Cottonseed: weigh 1g of each cottonwool and cottonseed kernel shell, accurate to
0.0001g;
---Cottonseed meal: weigh 1g, accurate to 0.0001g;
--- Other samples: weigh 5g, accurate to 0.01g.
7.1.2 Extraction of free gossypol
The sample was placed in a 250mL triangular flask with stopper; add 20 pieces of glass
beads; accurately add 50mL of solvent A; and the mixture is shaken and extracted at
room temperature at 120 times/min for 1h. Filter by filter paper; and cover the funnel
by a watch glass during filtering. Discard 5mL of the initial filtrate; and collect all the
remaining filtrate into a 100mL triangular flask with stopper for later-use. For
cottonseed samples, cottonwool and cottonseed kernel shells shall be placed in
different triangular flasks; and the other operations are the same as above.
7.2 Preparation of sample solution
7.2.1 Use a pipette to accurately pipette two aliquots of filtrate (7.1.2) of 5mL~10mL
(each filtrate contains about 50μg~100μg of free gossypol); and place them in two
25mL brown volumetric flasks a and b, respectively. If it is less than 10mL, use solvent
A to make up to 10mL.
7.2.2 Dilute a to the mark with isopropanol-n-hexane mixed solution (4.3); shake up;
this solution is used as a reference solution for the sample measurement solution.
7.2.3 Use a pipette to transfer two 10 mL of portions of solvent A into two 25 mL brown
volumetric flasks a0 and b0.
7.2.4 Dilute a0 to the mark by isopropanol-n-hexane mixed solution (4.3); shake up;
this solution is used as a reference solution for the blank measurement solution.
7.2.5 Add 2.0 mL of aniline (4.2) to b and b0; heat it on a boiling water bath for 30min;
then take it out; cool to room temperature; use isopropanol-n-hexane mixed solution
(4.3) to make the constant volume; shake well; and stand at room temperature for 1h.
7.3 Determination