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GB/T 40175.3-2021 English PDF (GBT40175.3-2021)

GB/T 40175.3-2021 English PDF (GBT40175.3-2021)

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GB/T 40175.3-2021: Textiles - Methods of biochemical analysis - Part 3: Organophosphorus pesticide (enzyme-linked immunosorbent assay)
This document specifies the enzyme-linked immunosorbent assay for four organophosphorus pesticide residues, including parathion-methyl, DIMETHYLP-NITROPHENYLPHOSPHATE, fenitrothion, and fenthion, in textiles. This document applies to all kinds of textiles.
GB/T 40175.3-2021
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 59.080.01
CCS W 04
Textiles - Methods of biochemical analysis - Part 3:
Organophosphorus pesticide (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 organophosphorus pesticide enzyme-
linked immunosorbent assay kit ... 10
References ... 11
Textiles - Methods of biochemical analysis - Part 3:
Organophosphorus pesticide (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 for four
organophosphorus pesticide residues, including parathion-methyl, DIMETHYL- P-NITROPHENYLPHOSPHATE, fenitrothion, and fenthion, 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 organophosphorus pesticides in the sample and the organophosphorus
pesticide antigens that have been coated on the ELISA plate in the kit
competitively combine with the organophosphorus pesticide antibody; then, add the enzyme-labeled secondary antibody to form an antigen-primary antibody- 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: Before use, mix substrate A solution 5.1e) and substrate B solution 5.1f) in a volume ratio of 1:1.
5.4 N-hexane.
5.5 Ethyl acetate.
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 1000 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: The sensitivities are 0.1 g and 0.000c1 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 1.0 g of sample from the mixed sample; place it in a 50 mL centrifuge Calculate the average absorbance value of the standard solution and the sample solution respectively; calculate the relative absorbance value of the standard solution or the sample solution according to Formula (1).
Where:
A – relative absorbance value;
B – average absorbance value of the series standard working solution or sample solution;
B0 – average absorbance value of 0 μg/L standard working solution.
8.2 Drawing of the standard working curve
Use the relative absorbance value as the ordinate (%) and the logarithm of the standard working solution concentration (μg/L) as the abscissa to draw a standard working curve. A standard working curve shall be drawn for each test. 8.3 Calculation and result expression of the concentration of
organophosphorus pesticide
From the standard working curve drawn in 8.2, read the concentration of organophosphorus pesticide corresponding to the relative absorbance of the sample solution. Calculate the content of organophosphorus pesticide in the sample according to Formula (2).
Where:
X – the content of organophosphorus pesticide in the sample, in micrograms per kilogram (μg/kg);
c – the concentration of organophosphorus pesticide in the sample, which is obtained from the standard working curve, in micrograms per liter (μg/L); V – the volume of the sample solution after reconstitution, in milliliters (mL); m – sample mass, in grams (g).
The calculation result shall be deducted from blanks and accurate to one decimal place.
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