GB/T 40152-2021Determination of saccharase activity in honey—Spectrophotometric method (English PDF)
蜂蜜中蔗糖转化酶的测定 分光光度法
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
May 21, 2021
Implementation date
December 1, 2021
Scope
GB/T 40152-2021 is the English-translated version of 蜂蜜中蔗糖转化酶的测定 分光光度法.
GB/T 40152-2021 gives a spectrophotometric method for measuring saccharase, or sucrose invertase, activity in honey.
The enzyme in a honey solution converts the substrate p-nitrophenyl-alpha-D-glucopyranoside into p-nitrophenol; after reaction at 40 °C for 20 minutes a stop solution ends it, and the absorbance read at 400 nm is directly proportional to the invertase value, with a blank control run alongside.
The standard lists the reagents and how to prepare them, including potassium dihydrogen phosphate, disodium hydrogen phosphate dihydrate, Tris, a 0.1 mol/L phosphate buffer at pH 6.0, a 0.02 mol/L substrate solution and a 3 mol/L stop solution, with water of the third grade in GB/T 6682.
Honey testing laboratories use it. It was issued on 21 May 2021 and has applied since 1 December 2021.
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Document preview — GB/T 40152-2021
National Standard of the People's Republic of China
- ICS
- 67.180.10
- Classification
- X 31
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- Foreword
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Principles of the method
- 5 Reagents and materials
Foreword
This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1: Structure and Drafting Rules of Standardization Documents" Drafting.
1 Scope
This document specifies the spectrophotometric method for the determination of sucrose invertase in honey.
This document is applicable to the determination of sucrose invertase in honey.
2 Normative references
The contents of the following documents constitute the indispensable clauses of this document through normative references in the text. Among them, dated quotations Only the version corresponding to that date is applicable to this document; for undated reference documents, the latest version (including all amendments) is applicable to this document.
GB/T 6682 Analytical laboratory water specifications and test methods
3 Terms and definitions
There are no terms and definitions that need to be defined in this document.
4 Principles of the method
The sucrose invertase contained in honey can quantitatively convert the substrate p-nitrobenzene-alpha-D-glucopyranoside into p-nitrophenol, and pass the test Determine the absorbance value at a specific wavelength to determine the content of p-nitrophenol to determine the sucrose invertase in honey. Add honey solution to substrate solution Mix well. After reacting at 40°C for 20 minutes, add a stop solution to terminate the reaction. Measure the absorbance at a wavelength of 400nm.
The absorbance value is directly proportional to the sucrose invertase value. At the same time, make a blank control.
5 Reagents and materials
Unless otherwise specified, the reagents used in this method are all analytically pure, and the water is the third grade water specified in GB/T 6682.
5.1 Potassium dihydrogen phosphate (KH2PO4).
5.2 Disodium hydrogen phosphate dihydrate (Na2HPO4·2H2O).
5.3 P-Nitrophenyl-alpha-D-glucopyranoside (CASNO. 3767-28-0).
5.4 Tris.
5.7 3mol/L stop solution.
5.5 0.1mol/L phosphate buffer (pH 6.0)
Weigh 11.66g potassium dihydrogen phosphate and 2.56g disodium hydrogen phosphate dihydrate (Na2HPO4·2H2O), dissolve in water, and dilute to 1L.
5.6 0.02mol/L substrate solution (pH 6.0)
Weigh 6.025g of p-nitrophenyl-alpha-D-glucopyranoside, add about 900mL of phosphate buffer (5.5) to dissolve, and if necessary, heat to Solubility at 60°C, cool to room temperature, dilute to 1L, store at 4°C in the dark, valid period 7d.
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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
Referenced standards
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