HPLC-MS를 이용한 생체시료 중 니세르골린의 주대사체인 10α-Methoxy-9,10-dihydrolysergol(MDL)의 분석 및 이를 이용한 한국인 성인 남성에 대한 생체이용률 응용

Determination of 10α-Methoxy-9,10-dihydrolysergol(MDL), Main Metabolite of Nicergoline, in Human Plasma by HPLC-MS and Applicability to Oral Bioavailability in Korean Heathy Male Volunteers

A simple and sensitive HPLC-MS method for quantitation of 10α-Methoxy-9,10-dihydrolysergol(MDL), the main metabolite of nicergoline, in human plasma was developed and the bioavailability parameters of MDL was assessed in Korean healthy male volunteers. Clomipramine was used as an internal standard. MDL and internal standard in plasma sample were extracted using ethyl acetate. A centrifuged upper layer was then evaporated and reconstituted with mobile phase of 10 mM ammonium acetate-acetonitrile (10:90, v/v). The reconstituted samples were injected into a Zorbax SB-C8 column (2.1×150mm, 5μM) at a flow-rate of 0.3㎖/min. Using MS with selected ion monitoring (SIM) mode, MDL and clomipramine were detected without sere interference from human plasma matrix. MDL produced a protonated molecular ion ([M+H]+) at m/z 287. Internal standard produced a protonated molecular ion ([M+H]+) at m/z 315. A linear relationship for MDL was found in the range of 2.5~100ng/㎖. The lower limit of quantitation (LLOQ) was 2.5ng/㎖ with acceptable precision and accuracy. The intra- and inter-day validation for all coefficients of variation (R.S.D.%) were found less than 15%. Main pharmacokinetic parameters of 30mg of nicergoline were revealed as follow: AUCt 321.1±64.5ng․hr/㎖, Cmax 51.2±25.3ng/㎖, Tmax 3.6±1.5hr, Kel0.12±0.07hr-1 and t1/2 7.6±3.4 hr. Inter subject variations and race differences were shown in comparison with the published data in the literature.