GB/T 40980-2021 in English

VALID

Determination of reducing sugar in biochemical products—High-performance liquid chromatography with pre-column derivatization

Issued on:2021-11-26
Implemented on:2022-06-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $150.00
$146.00

Standard No:	GB/T 40980-2021
Document status:	VALID
Superseded by:	GB/T 40980-2026 Determination of reducing sugars in biochemical products
Superseded on:	2026-10-01
Title in English:	Determination of reducing sugar in biochemical products—High-performance liquid chromatography with pre-column derivatization
Title in Chinese:	生化制品中还原糖的测定柱前衍生高效液相色谱法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2021-11-26
Implemented on:	2022-06-01
ICS Classification:	07.080-Biology. Botany. Zoology
Chinese Classification:	A40-Basic Subject in general
Professional Classification:	GB-National Standard
Related Keywords:	pre-column derivatization introduction technical background biochemical products high-performance liquid chromatography derivatization time derivatization efficiency biochemical products
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《GB/T 40980-2021生化制品中还原糖的测定柱前衍生高效液相色谱法》由TC387（全国生化检测标准化技术委员会）归口，主管部门为国家标准化管理委员会。

Introduction

Technical background of the standard

This standard aims at the common needs of reducing sugar detection in biochemical products, and innovatively adopts PMP pre-column derivatization-HPLC technology to solve the problems of poor selectivity and time-consuming and labor-intensive traditional DNS method. The method development took 3 years and was jointly verified by 8 laboratories, with a precision of ±10%.

Principle of the core method

Steps	Key parameters	Mechanism of action
Derivatization reaction	70℃/60min	PMP condenses with the reducing sugar aldehyde group to form a stable derivative
Purification treatment	Chloroform extraction 3 times	Remove interference from excess derivatization reagents
Chromatographic separation	pH7.8 phosphate gradient	Achieve baseline separation of 10 sugars

Key operating points

Case: When a vaccine manufacturer tested the glucose content in human immunoglobulin, it was found that the derivatization efficiency was insufficient. After investigation, it was found that:

The water bath temperature did not meet the standard requirement of 70℃±1℃
Nitrogen drying was not thorough and HCl remained

After correction, the measured recovery rate increased from 82% to 98%.

Method performance comparison

Indicators	This method	DNS method	Enzyme method
Detection limit (μg/mL)	0.19-0.45	5-10	1-2
Analysis time	90min	180min	30min
Sugar types	10 types	Total amount	Monosaccharide

Implementation suggestions

Quality control points: Each batch of samples needs to be subjected to blank test and spike recovery simultaneously (control range 85-110%)
Chromatographic column maintenance: Immediately rinse with 10% acetonitrile after use to avoid phosphate crystallization
Derivatization optimization: For disaccharides such as lactose, the derivatization time can be extended to 90min

More samples: DL/T 5154-2012 JGJ 94-2008 GB 50021-2001(2009)

Sample only — not a preview of GB/T 40980-2021

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