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GB/T 40175.2-2021 English PDF (GBT40175.2-2021)

GB/T 40175.2-2021 English PDF (GBT40175.2-2021)

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GB/T 40175.2-2021: Textiles - Methods of biochemical analysis - Part 2: Pyrethroid pesticides(enzyme-linked immunosorbent assay)

This document specifies the enzyme-linked immunosorbent assay method for the detection of residues of seven pyrethroid pesticides, including fenpropathrin, deltamethrin, cypermethrin, beta-cypermethrin, cyfluthrin, RU 38702, and cyphenothrin, in textiles. This document applies to all kinds of textiles.
GB/T 40175.2-2021
GB
NATIONAL STANDARD OF THE
PEOPLE REPUBLIC OF CHINA
ICS 59.080.01
CCS W 04
Textiles - Methods of biochemical analysis - Part 2:
Pyrethroid pesticides (enzyme-linked immunosorbent
assay)
ISSUED ON: MAY 21, 2021
IMPLEMENTED ON: DECEMBER 01, 2021
Issued by: State Administration for Market Regulation;
Standardization Administration of the PEOPLE Republic of
China.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Terms and definitions ... 4
4 Principle ... 4
5 Reagents and materials ... 5
6 Equipment and utensils ... 6
7 Test procedure ... 6
8 Calculation and expression of results ... 7
9 Detection limit ... 9
10 Test report ... 9
Appendix A (Informative) Example of pyrethroid pesticide enzyme-linked
immunosorbent assay kit ... 10
References ... 11
Textiles - Methods of biochemical analysis - Part 2:
Pyrethroid pesticides (enzyme-linked immunosorbent
assay)
WARNING: The personnel who uses this document shall have hands-on
experience in formal laboratory work. This document does not address all possible security issues. It is the responsibility of the user to take
appropriate safety and health measures and to ensure compliance with
the conditions which are set by the relevant national regulations.
1 Scope
This document specifies the enzyme-linked immunosorbent assay method for the detection of residues of seven pyrethroid pesticides, including fenpropathrin, deltamethrin, cypermethrin, beta-cypermethrin, cyfluthrin, RU 38702, and cyphenothrin, in textiles.
This document applies to all kinds of textiles.
2 Normative references
The contents of the following documents constitute the indispensable clauses of this document through normative references in the text. For dated references, only the version corresponding to that date is applicable to this document; for undated references, the latest version (including all amendments) is applicable to this document.
GB/T 6682, Water for analytical laboratory use - Specification and test methods
3 Terms and definitions
There are no terms and definitions that need to be defined in this document. 4 Principle
The pyrethroid pesticides in the sample and the pyrethroid pesticide antigens that have been coated on the microplate in the kit competitively combine with the pyrethroid pesticide antibody; then, add the enzyme-labeled secondary 5.2 Washing liquid: Before use, add water to dilute the concentrated washing solution 5.1h) in a volume ratio of 1:9 (1 part of concentrated washing solution + 9 parts of water).
5.3 Color-developing agent: Mix substrate A solution 5.1e) and substrate B solution 5.1f) in a volume ratio of 1:1 before use.
5.4 N-hexane.
5.5 Petroleum ether.
5.6 Methanol.
5.7 Phosphate buffer solution (0.01 mol/L): Dissolve 8.0 g of sodium chloride, 0.2 g of potassium chloride, 1.44 g of disodium hydrogen phosphate and 0.24 g of potassium dihydrogen phosphate in 800 mL of deionized water; use 0.01 mol/L hydrochloric acid to adjust the pH value of the solution to 7.4; add water to make the volume up to 1 000 mL.
5.8 Methanol-phosphate buffer solution: Mix methanol and buffer solution (5.7) in a volume ratio of 1:4.
6 Equipment and utensils
6.1 Microplate reader: The wavelength is 450 nm.
6.2 Balance: sensitivity of 0.1 g and 0.000 1 g.
6.3 Micro adjustable pipette and matching tips: 50 ??L, 100 ??L, 1 000 ??L. 6.4 Temperature-controllable ultrasonic bath: The working frequency is 40 kHz; the temperature control accuracy is ??5 ??C.
6.5 Nitrogen-blowing instrument.
6.6 Oven: The temperature control accuracy is ??1 ??C.
6.7 pH meter: It is equipped with glass electrode; the measurement accuracy is at least accurate to 0.1.
7 Test procedure
7.1 Sample pretreatment
Weigh 5 g of the representative sample; cut it to 5 mm ?? 5 mm or less; mix well. Weigh about 1.0 g of sample from the mixed sample; place it in a 50 mL
centrifuge tube with a screw cap. Accurately add 20 mL of a mixed solvent of n- hexane (5.4) and petroleum ether (5.5) at a volume ratio of 1:1; place it in an ultrasonic bath (6.4) at 45 ??C for extraction for 30 minutes; take 1 mL of the extract; use nitrogen to dry it; use 100 ??L of methanol-phosphate buffer solution (5.8) to redissolve for test.
7.2 Enzyme-linked immunosorbent assay
7.2.1 Before testing, all reagents need to be placed at room temperature (20 ??C ~ 25 ??C) before they can be used.
7.2.2 Take 50 ??L of the series fenpropathrin standard working solution 5.1d) and 50 ??L of the to-be-tested sample solution (7.1) respectively; add them to the corresponding hole of the microtiter plate 5.1a); test 2 of each solution in parallel; then, add 50 ??L of pyrethroid antibody working solution 5.1b) respectively; let them react in an oven (6.6) at 37??C for 30 minutes.
7.2.3 Discard the liquor in the hole; pat dry the remaining liquid of the microtiter plate 5.1a) on the absorbent paper. Fill each hole with washing liquid (5.2); shake gently; leave it for 2 minutes; discard the liquid in the hole; pat dry on absorbent paper; repeat the washing 4 times.
7.2.4 Add 50 ??L of enzyme-labeled secondary antibody 5.1c) to each hole; react in an oven (6.6) at 37??C for 30 minutes.
7.2.5 Discard the liquor in the hole; pat dry the remaining liquor of the microtiter plate on the absorbent paper; fill each hole with washing liquid (5.2); shake gently; leave it for 2 minutes; discard the liquid in the hole; pat dry on absorbent paper; repeat the washing 4 times.
7.2.6 Add 50 ??L of color-developing agent (5.3) to each hole; develop color in an oven (6.6) at 37??C in the dark for 15 minutes.
7.2.7 Add 50 ??L of stop solution (5.1g) to each hole to stop the reaction; use a microplate reader (6.1) of a wavelength of 450 nm to measure the absorbance value within 10 minutes.
7.3 Blank test
In the case of no sample, carry out the test according to the above steps. 8 Calculation and expression of results
8.1 Calculation of relative absorbance value

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