L-02细胞中3-羟基-3-甲基戊二酰辅酶A还原酶活性检测方法的建立及应用  

Establishment and application of detection method for determinating the activity of 3-hydroxy-3-methyl glutaric acyl coenzyme A reductase in L-02 cells

作　　者：孙旭[1,2] 罗余萍[2] 熊玉卿[2] 

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心/药物非临床安全评价研究北京市重点实验室,北京100176 [2]南昌大学医学院临床药理研究所,南昌330006

出　　处：《中国临床药理学杂志》2015年第20期2046-2048,共3页The Chinese Journal of Clinical Pharmacology

摘　　要：目的建立L-02细胞中3-羟基-3-甲基戊二酰辅酶A(HMG-Co A)还原酶活性的检测方法。方法色谱柱:C18色谱柱,流动相:甲醇-水(20μmol·L-1磷酸二氢钾,p H 7.2)=6∶94,检测波长:340 nm,柱温:25℃,流速:1m L·min-1。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性。结果还原型辅酶Ⅱ线性范围为2.5~1000μmol·L-1,定量下限为0.5μmol·L-1,日内、日间RSD分别在(5~6)%和(4~8)%内,相对回收率在(96.97~99.57)%内。结论建立的L-02细胞中HMG-Co A还原酶活性检测方法符合实验要求,且可用于测定氟伐他汀对L-02细胞HMG-Co A还原酶活性的抑制作用。Objective To establish a method of determination the activi- ty of 3 -hydroxy- 3 -methyl glutaric acyl coenzyme A reductase in L-02 cells. Methods Chromatographic column was Cls chromato- graphic column. Mobile phase was methanol -water （20 p^mol ~ L-l po- tassium dihydrogen phosphate, pH 7.2） = 6： 94. Detection wavelength was 340 nm. Column temperature was 25 ~C . Flow rate was 1 mL. min-1 The specificity, standard curve and quantitative limit, precision and recovery, and stability were investigated. Results The concentration range of nicotinamide adenine dinucleotide phosphate was 2. 5 -1000 p, mol~ L-1 with a good linear relationship. Lower limit of quantitation was 0.5 p, mol ~ L- 1. Intra - and inter - day RSD were （ 5 - 6） % and （ 4 - 8 ） %, respectively. Relative recovery was （ 96. 97 - 99.57） %. Conclusion The established detection method for activity de- termination of HMG - CoA reductase in L - 02 cells complied with experi- mental requirements, which could be used for the determination of inhibitory effect of iluvastatin on the activity of HMG - CoA reductase in L - 02 cells.

关 键 词：3-羟基-3-甲基戊二酰辅酶A还原酶活性 L-02细胞 高效液相色谱 还原型辅酶Ⅱ 氟伐他汀 

分 类 号：R965.2[医药卫生—药理学] R966[医药卫生—药学]