HPLC检测人重组组织因子脂化物磷脂含量  

Quantification of phospholipids forming a liposome by high performance liquid chromatography

作　　者：赵生旺 梁欣欣 杨欣 赵峰梅 赵邑 彭秀英 ZHAO Sheng-wang;LIANG Xin-xin;YANG Xin;ZHAO Feng-mei;ZHAO Yi;PENG Xiu-ying(Shanxi Key Laboratory of pharmaceutical Biotechnology,Shanxi Biological Research Institute Co.Ltd.,Taiyuan 030006,China;Department of Environment and Safety Engineering,Taiyuan Institute of Technology,Taiyuan 030008,China)

机构地区：[1]山西省生物研究院有限公司,医药生物技术山西省重点实验室,山西太原030006 [2]太原工业学院环境与安全工程系,山西太原030008

出　　处：《化学研究与应用》2023年第1期53-58,共6页Chemical Research and Application

基　　金：山西省基础研究计划项目(202103021223351,201901D211597)资助。

摘　　要：利用脂质体技术制备凝血酶原时间测定试剂人重组组织因子脂化物,建立高效液相色谱(HPLC)方法用于同时分析人重组组织因子脂化物中的主要成分磷脂酰丝氨酸(DOPS),磷脂酰胆碱(DOPC),以及人重组组织因子(TF)的半定量测定。分析测定条件:色谱柱:Waters Symmetry300 C4色谱柱(150×4.6 mm,5μm,300),流动相甲醇水,梯度洗脱,流速1 mL·min^(-1),柱温:30℃,检测波长210 nm,DOPS保留时间12.275 min,TF保留时间15.4 min,DOPC保留时间18.917 min。研究发现,采用建立的分析方法,DOPS在1.875~15 mg·mL^(-1)的浓度范围内呈良好的线性关系(R 2=0.9974),检出限为1.875 mg·mL^(-1)(S/N=3);DOPC在4.375~35 mg·mL^(-1)浓度范围内线性关系(R 2=0.9993)良好,检出限为4.375 mg·mL^(-1)(S/N=3)。在结合多次制备的人重组组织因子脂化物的产品测试中,该方法也表现出了良好的稳定性和精密度。总之,该方法可同时测定脂化物成分组成,并具有良好的检测效果,可为化妆品、制药和食品补充剂行业提供理想的参考价值。Based on the liposomal technology of encapsulation for the preparation of human recombinant tissue factor as the prothrombin time assay reagent,a high-performance liquid chromatography(HPLC)method was developed to simultaneously analysis three widely used active substances such as phosphatidylserine(DOPS),phosphatidylcholine(DOPC),and tissue factor(TF).In this new approach,the optimization conditions were found including current timing and costs in the analyses of these active substances,with the mobile phase comprising of methanol flowing at 1.0 mL·min^(-1) in a gradient elution manner,and the detection wavelength was set at 210 nm.The chromatographic column of Waters Symmetry300 C4(150×4.6 mm,5μm,300)was used for analysis.With constrained optimization,DOPS showed good linear relationships in a range of 1.875~15 mg·mL^(-1) with a LOD of 1.875 mg·mL^(-1)(S/N=3),and DOPC with a linear range of 4.375~35 mg·mL^(-1) and a LOD of 4.375 mg·mL^(-1)(S/N=3).Through the actual sample determination,the developed method was demonstrated by the promising results on the simultaneous analyses of DOPS and DOPC.Thus,this method could result in an ideal analytic method for the cosmetics,pharmaceutical,and food supplement industry.

关 键 词：高效液相色谱 人重组组织因子脂化物 磷脂酰胆碱 磷脂酰丝氨酸 

分 类 号：O657.7[理学—分析化学]