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日用化学工业（中英文）

日用化学工业 ›› 2017, Vol. 47 ›› Issue (11): 659-662.doi: 10.13218/j.cnki.csdc.2017.11.013

• 分析与检测 • 上一篇    

高效液相色谱-蒸发光散射法测定蛇皮中胶原蛋白含量

丁瑞敏1,2, 周华锋2,3, 李赟高3, 蔡衡3, 段明星2, 靳清贤1   

  1. 1.郑州轻工业学院 材料与化学工程学院,河南 郑州 450001;
    2.清华大学 生命科学学院,北京 100084;
    3.江苏隆力奇生物科技股份有限公司,江苏 常熟 215500
  • 收稿日期:2017-08-12 出版日期:2017-11-22 发布日期:2019-03-21
  • 通讯作者: 周华锋,高级工程师,电话:13426015613,E-mail:zhf031689@163.com。靳清贤,讲师,电话:18203684694,E-mail:13253525730@163.com。
  • 作者简介:丁瑞敏(1992-),女,河南开封人,硕士研究生,电话:18801065578,E-mail:dingrm0422@126.com。

Determination of collagen content in snake skin by HPLC-ELSD

DING Rui-min1,2, ZHOU Hua-feng2,3, LI Yun-gao3, CAI Heng3, DUAN Ming-xing2, JIN Qing-xian1   

  1. 1.Department of Material and Chemical Engineering,Zhengzhou University of Light Industry,Zhengzhou, Henan 450001,China;
    2.School of Life Sciences,Tsinghua University,Beijing 100084,China;
    3.Jiangsu Longliqi Biotechnology Limited Company,Changshu,Jiangsu 215500,China
  • Received:2017-08-12 Online:2017-11-22 Published:2019-03-21

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摘要： 建立高效液相色谱-蒸发光散射检测(HPLC-ELSD)法测定蛇皮胶原蛋白含量的方法。胶原蛋白经盐酸水解后,采用Thermo Hypersil Gold C18(250 mm×4.6 mm,5 μm)色谱柱进行分离,以乙腈(含0.2%甲酸,A相)-0.2%甲酸水溶液(B相)为流动相进行梯度洗脱,洗脱时间15 min,柱温30 ℃,流速0.4 mL/min。ELSD参数:雾化器温度85 ℃,蒸发器温度85 ℃,氮气作载气,蒸发器气流1.2 L/min。进样量10 μL。在此条件下,羟脯氨酸的线性范围为10~100 mg/L,R2=0.999 6,检测限为5.56 mg/L,定量限为10 mg/L。蛇皮胶原蛋白的平均回收率为94.53%,RSD为2.15%。蛇皮中胶原蛋白质量分数平均值为40.21%,为蛇皮的主要蛋白质。

关键词: 化妆品原料, 胶原蛋白, 羟脯氨酸, 高效液相色谱-蒸发光散射法, 蛇皮

Abstract: A high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method for determination of content of collagen in snake skin was established.After the collagen in snake skin sample was hydrolyzed with hydrochloric acid,separation was performed by a Thermo Hypersil Gold C18(250 mm×4.6 mm,5 μm) chromatographic column and acetonitrile (containing 0.2% formic acid,mobile phase A)-water (containing 0.2% formic acid,mobile phase B) as the mobile phase were used for gradient elution.The elution time is 15 min.The column temperature is 30 ℃;and the flow rate is 0.4 mL/min.The ELSD detector parameters are as follows:nebulization temperature,85 ℃;evaporation temperature,85 ℃;carrier gas,N2;air flow of the evaporator,1.2 L/min.The injection volume of the sample is 10 μL.Under these chromatographic conditions,the linearity range of hydroxyproline is 10 - 100 mg/L,R2 = 0.999 6;the lowest limit of detection is 5.56 mg/L,and the lowest limit of quantification is 10 mg/L.The average recovery rate of snake skin collagen is 94.53%;the RSD is 2.15%;and the average of the mass fraction of collagen in snake skin is 40.21%.

Key words: cosmetic ingredient, collagen, hydroxyproline, HPLC-ELSD, snake skin

中图分类号: 

  • TQ658