Page 110 - 《中国医疗器械杂志》2025年第6期

Chinese Journal of Medical Instrumentation 2025年 第49卷 第6期

监 管 与 测 试

文章编号：1671-7104(2025)06-0696-07

高效液相色谱法测定重组人源化胶原蛋白敷料中

胶原蛋白含量

【作 者】 徐文慧，何振瑞，岳海霞，范玉洁
山西锦波生物医药股份有限公司，太原市，030032
【摘 要】 目的 建立一种高效液相色谱法用于定量检测重组人源化胶原蛋白敷料中重组Ⅲ型人源化胶原蛋白的含量，
对该胶原蛋白以及相关产品进行质量控制。方法 采用高效液相色谱法，参数：Agilent Eclipse XDB-C18色
谱柱（4.6×150 mm, 5 μm）；以0.1%三氟乙酸水溶液和0.1%三氟乙酸乙腈溶液为流动相；柱温30 ℃；
流速1.0 mL/min；检测波长214 nm。结果 该法专属性强。对重组Ⅲ型人源化胶原蛋白含量＞0.1 mg/mL
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的敷料用水解法处理后，在0.1~1.2 mg/mL的范围内其浓度与峰面积具有良好的线性关系（R =0.999 4），
加标平均回收率在90.91%~94.14%，检出限和定量限分别为0.05、0.1 mg/mL。对重组Ⅲ型人源化胶原蛋
白含量≤0.1 mg/mL的敷料用酶解法处理后，在0.05~1.0 mg/mL的范围内其浓度与峰面积具有良好的线性关
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系（R =0.999 5），可以用来指导低浓度敷料蛋白的检测。结论 该方法快速、准确、重复性好，可以用来
检测敷料中胶原蛋白的含量。
【关 键 词】 高效液相色谱法；重组Ⅲ型人源化胶原蛋白；质量控制
【中图分类号】 TQ937
【文献标志码】 A doi: 10.12455/j.issn.1671-7104.250207
Determination of Collagen Content in Recombinant Human Collagen
Dressing by High Performance Liquid Chromatography

【 Authors 】 XU Wenhui, HE Zhenrui, YUE Haixia, FAN Yujie
Shanxi Jinbo Biotechnology Co., Ltd., Taiyuan, 030032
【 Abstract 】 Objective To establish a high performance liquid chromatography method for the determination of
recombinant type Ⅲ human collagen in recombinant human collagen dressing for quality control of the
product. Methods HPLC was utilized, and its parameters were as follows: an Agilent Eclipse XDB-C18
column (4.6×150 mm, 5 μm) was used with 0.1% TFA solution and 0.1% TFA acetonitrile solution as
mobile phase at 30 ℃, the flow rate was 1.0 mL/min, and the detection wavelength was 214 nm. Results
This method demonstrates high specificity. For dressings with a recombinant type Ⅲ human collagen
content greater than 0.1 mg/mL, after hydrolysis treatment, a good linear relationship between
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concentration and peak area was observed within the range of 0.1 to 1.2 mg/mL (R =0.999 4). The
average spiked recovery ranged from 90.91% to 94.14%, with a detection limit (LOD) of 0.05 mg/mL and
a quantification limit (LOQ) of 0.1 mg/mL. For dressings with a recombinant type Ⅲ human collagen
content of 0.1 mg/mL or less, after enzymatic digestion treatment, a good linear relationship between
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concentration and peak area was obtained within the range of 0.05 to 1.0 mg/mL (R =0.999 5). This
validates the method’s suitability for guiding the detection of low-concentration dressing proteins.
Conclusion The method is rapid, accurate and reproducible, and can be used to determine the content of
type Ⅲ collagen in dressings.
【Key words】 high performance liquid chromatography (HPLC), recombinant type III human collagen, quality control

0 引言 的皮肤、骨、软骨、肌腱、韧带等组织中，是结缔
组织的重要结构蛋白质，也是细胞外基质中最重要
胶原蛋白是一种生物高分子，广泛存在于动物 的组成部分，不仅拥有机械稳定性，更有促进细胞

[1]
生长和迁移的能力 。在人体皮肤细胞的外层基质
收稿日期：2025-03-29
作者简介：徐文慧，E-mail: 1067950089@qq.com 中，胶原蛋白占80%。它是人体内数量最庞大、含

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