高效液相色谱-蒸发光散射法测定蛇皮中胶原蛋白含量  

Determination of collagen content in snake skin by HPLC-ELSD

作　　者：丁瑞敏 周华锋 李赟高[3] 蔡衡[3] 段明星 靳清贤[1] 

机构地区：[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

出　　处：《日用化学工业》2017年第11期659-662,共4页China Surfactant Detergent & Cosmetics

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,5μm)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 m L/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10μL。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。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 is0. 4 m L/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 is10 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%.

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 

分 类 号：O657[理学—分析化学] R284.1[理学—化学]