CAJ | 학술논문

目的：建立测定人体肝脏组织中CYP3 A4酶活性的HPLC方法。方法：取21例肝癌、16例肝血管瘤、13例胆组织病变患者的肝组织，制作肝微粒体，体外孵育后以咪达唑仑（ MDZ）为探针药物，用高效液相色谱法测定CYP3A4的Km值。 HPLC色谱条件：色谱柱为Angel C-18柱（4．6 mm ×250 mm，5μm），流动相为甲醇：乙腈：水（体积比53砄13砄34），流速1 mL/min，检测波长220 nm，柱温25℃，进样量40μL。结果：1’-OH MDZ与MDZ的出峰时间分别为10．05与15．02 min，分离充分（R＞1．5），1’-OH MDZ质量浓度的范围为200～6000μg/L，高、中、低浓度回收率均＞90％，日内、日间精密度RSD均＜5％，稳定性RSD均＜5％。50例患者Km值1．75～30．17μmol/L；不同性别、年龄及肝胆病变患者的Km值差异无统计学意义（P＞0．05）。结论：该法稳定、快速、重复性好，可用于人体肝组织中CYP3A4酶活性的测定；性别、年龄及肝胆病变情况对CYP3A4酶活性无影响。
목적：건립측정인체간장조직중CYP3 A4매활성적HPLC방법。방법：취21례간암、16례간혈관류、13례담조직병변환자적간조직，제작간미립체，체외부육후이미체서륜（ MDZ）위탐침약물，용고효액상색보법측정CYP3A4적Km치。 HPLC색보조건：색보주위Angel C-18주（4．6 mm ×250 mm，5μm），류동상위갑순：을정：수（체적비53결13결34），류속1 mL/min，검측파장220 nm，주온25℃，진양량40μL。결과：1’-OH MDZ여MDZ적출봉시간분별위10．05여15．02 min，분리충분（R＞1．5），1’-OH MDZ질량농도적범위위200～6000μg/L，고、중、저농도회수솔균＞90％，일내、일간정밀도RSD균＜5％，은정성RSD균＜5％。50례환자Km치1．75～30．17μmol/L；불동성별、년령급간담병변환자적Km치차이무통계학의의（P＞0．05）。결론：해법은정、쾌속、중복성호，가용우인체간조직중CYP3A4매활성적측정；성별、년령급간담병변정황대CYP3A4매활성무영향。
Aim:To establish a HPLC method for determination of CYP 3A4 activity in human hepatic tissue .Meth-ods:Human liver microsome of 21 cases of liver cancer ,16 cases of hepatic hemangioma ,and 13 cases of gallbladder lesion was prepared for HPLC analysis .The analytical column was Angel C-18 column (4.6 mm ×250 mm, 5 μ), mobile phase consisted of methanol-acetonitrile-water( volume ratio =53:13:34 ) with flow rate at 1 mL/min, detective wave-length at 220 nm, and column temperature at 25℃,and the injection volume was 40μL..Results:1’-OH MDZ and MDZ retention time was 10.05 and 15.02 min with symmetrical peak and full separation (R>1.5) .The linear range of 1'-OH MDZ was 200~6 000 μg/L.Extraction recoveries at low , medium and high concentrations were all higher than 90%, re-spectively .The intra-day and inter-day precision RSD was less than 5%, and the stability RSD was less than 5%.The sta-tistical analysis showed that the liver diseases, gender or age had no significant effect on CYP3A4 activity(P>0.05). Conclusion:The HPLC method is rapid, stable with high repeatability.It is suitable for the determination of CYP3A4 ac-tivity in human liver .The liver diseases , gender and age have no significant effects on the Km.