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GB/T 44828-2024Assay method of glucose oxidase activity (English PDF)

葡萄糖氧化酶活性检测方法

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Issued by

SAMR; SAC

Level / Type

National · Recommended

Issue date

October 26, 2024

Implementation date

October 26, 2024

Scope

GB/T 44828-2024 is the English-translated version of 葡萄糖氧化酶活性检测方法.

GB/T 44828-2024 is the Chinese national standard covering how much glucose oxidase activity a preparation actually has — the principle, in which the peroxide released from beta-D-glucose colours o-dianisidine, the reagents and the buffer, the test procedure and the rate at which absorbance rises at 500 nm, and the quality control, without which two laboratories quoting an activity in units for the same preparation are not quoting the same number. An enzyme is bought by activity rather than by weight, so the assay is what the price rests on. First edition, in force from 26 October 2024. Issued on 26 October 2024, it has been in force since 26 October 2024.

Document preview — GB/T 44828-2024

National Standard of the People's Republic of China

ICS: 07.080
Classification: C 27

Issued by: State Administration for Market Regulation; Standardization Administration of the PRC

Foreword3
1 Scope4
2 Normative References4
3 Terms and Definitions4
4 Principle4
5 Reagents or Materials5
6 Test Procedures5
7 Quality Control7

Foreword

This document was drafted in accordance with the rules provided in GB/T 1.1-2020 Directives for Standardization - Part 1: Rules for the Structure and Drafting of Standardizing Documents.

Please be noted that certain content of this document may involve patents. The issuing institution of this document does not undertake the responsibility of identifying these patents.

This document was proposed by and shall be under the jurisdiction of National Technical Committee 11 on Research Use Enzyme of Standardization Administration of China (SAC/SWG 11).

The drafting organizations of this document. Nansheng (Xiamen) Analytical Testing Co., Ltd.;

Center Testing International Co., Ltd.; Ningxia Sunson Industrial Group Co., Ltd.; Shandong Provincial Laboratory of Weihai Advanced Medical Materials and High-end Medical Devices;

Xiamen Yin Xiang Group Co., Ltd.; Shenzhen New Industries Biomedical Engineering Co., Ltd.; Wuhan SunHY Biology Co., Ltd.; Langfang Nuodao Zhongke Medical Laboratory Co., Ltd.; Shenzhen Heto Medical Tech Co., Ltd.; Tianjin Bofeide Technology Co., Ltd.; Guangzhou Jinde Biotechnology Co., Ltd.; Shanxi Dayu Bioengineering Co., Ltd.; Hebei Research Institute of Microbiology Co., Ltd.; Wuhan Hzymes Biotechnology Co., Ltd.; Shenzhen Huatongwei International Inspection Co., Ltd.; Hangzhou Assure Tech Co., Ltd.; Guangdong Cardlo Biotechnology Co., Ltd.; Bolido Biotechnology (Shijiazhuang) Co., Ltd.; SAHO (Shenzhen) Biotechnology Co., Ltd.; SAHO (Hangzhou) Biotechnology Co., Ltd.; Fujian Nansheng Technology Co., Ltd.; Fudan University; Shandong University; Soochow University; Xiamen University; Fujian Agriculture and Forestry University; Xiamen Huaxia University.

The main drafters of this document. Huang Facan, Zheng Dengzhong, Bai Honghai, Shen Tao, Zhang Zhigang, Ma Fuqiang, Du Kai, Xu Li, Pang Tieliang, He Yi, Wang Pengyuan, Li Minyou, Zhang Guangmin, Ma Qinghe, Shi Jingni, Yu Liangqing, Chen Dong, Xu Sheng, Zhang Gengbo, Zhao Yi, Guo Yanwei, Lin Yanna, Wang Tianhua, Zheng Tianye, Huang Enming, Tang Lin, Pan Wei, Zhong Jiang, Chen Xiulan, Zhu Li, Zhang Yongyou, Liu Bin, Zhao Chao, Zhou Zhongai, Pan Paifeng, Huang Haiyan.

Assay Method of Glucose Oxidase Activity

1 Scope

This document describes the assay method of glucose oxidase activity.

This document applies to the assay of glucose oxidase activity.

2 Normative References

GB/T 6682

3 Terms and Definitions

The following terms and definitions are applicable to this document.

3.1 Glucose oxidase

Under the condition where there is oxygen, a dehydrogenase that specifically catalyzes beta-D- glucose to generate D-gluconic acid and hydrogen peroxide.

3.2 Activity unit of glucose oxidase

At pH 5.1 and 35 °C, the amount of enzyme required to oxidize 1.0 mol of beta-D-glucose to D- gluconic acid and hydrogen peroxide per minute is defined as one activity unit (U).

4 Principle

Under the action of glucose oxidase, glucose reacts with oxygen to generate gluconic acid and hydrogen peroxide. Hydrogen peroxide reacts with colorless reduced o-dianisidine under the action of peroxidase to generate water and red oxidized o-dianisidine. This red substance has a specific absorption peak at a wavelength of 500 nm. By determining the rate of increase in absorbance at 500 nm of the red substance, the number of activity units of the enzyme can be calculated.

5 Reagents or Materials

5.1 Water

Grade-2 water conforming to the provisions of GB/T 6682.

5.2 50 mmol/L Sodium Acetate Buffer Solution Weigh-take 0.82 g of anhydrous sodium acetate and use 150 mL of pure water to dissolve it. At 35 °C, use 1 mol/L hydrochloric acid to adjust the pH to 5.1 and reach a constant volume of 200 mL. 5.3 0.21 mmol/L o-Dianisidine Weigh-take 50 mg of o-dianisidine dihydrochloride and use 7.5 mL of pure water to dissolve it.

Take 1 mL and use 50 mmol/L sodium acetate buffer solution (pH 5.1) to dilute it and reach a constant volume of 100 mL.

5.4 10% beta-D(+) Glucose Solution Weigh-take 1 g of beta-D(+) glucose, use pure water to dissolve it and reach a constant volume of 10 mL.

5.5 0.17 mmol/L o-Dianisidine-1.72% Glucose Solution Take 24 mL of 0.21 mmol/L o-Dianisidine and 5 mL of 10% -D(+) glucose solution, mix well, and keep it at a constant temperature of 35 C for later use. Prepare it right before use.

5.6 Horseradish Peroxidase Solution

Take an appropriate amount of horseradish peroxidase, use ice pre-cooled pure water to prepare it and dilute it to a solution of 60 red phenol units per mL (U/mL).

5.7 Test Sample Solution

Weigh-take an appropriate amount of the test sample solid or solution, use ice pre-cooled 50 mmol/L sodium acetate buffer solution (pH 5.1) to dissolve it and dilute to a solution of 0.4 U/mL ~ 0.8 U/mL.

6 Test Procedures

6.1 Substrate Solution

In accordance with Table 1, prepare the substrate working solution, and maintain it in a water bath at 35 C for 5 min. ......

......
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

Normative references

GB/T 6682Water for analytical laboratory use - Specification and test methods

Editions of GB/T 44828

Edition	Title	Revision	Status
GB/T 44828-2024	Assay method of glucose oxidase activity	current edition	Current

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