GB/T 40175.2-2021 PDF in English
GB/T 40175.2-2021 (GB/T40175.2-2021, GBT 40175.2-2021, GBT40175.2-2021)
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Textiles - Methods of biochemical analysis - Part 2: Pyrethroid pesticides(enzyme-linked immunosorbent assay)
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Standards related to: GB/T 40175.2-2021
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GB/T 40175.2-2021: PDF in English (GBT 40175.2-2021) GB/T 40175.2-2021
GB
NATIONAL STANDARD OF THE
PEOPLE’S 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’s 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 μL, 100 μL, 1 000 μL.
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 μL 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 μL of the series fenpropathrin standard working solution 5.1d)
and 50 μL 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 μL 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 μL 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 μL 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 μL 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
......
Source: Above contents are excerpted from the PDF -- translated/reviewed by: www.chinesestandard.net / Wayne Zheng et al.
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