LC-MS/MS法测定大鼠血浆中DPHC浓度及药动学应用

Determination of DPHC in Rat Plasma by LC-MS/MS and Its Application in Pharmacokinetics Study

目的建立快速、灵敏测定大鼠血浆中5-羟基-1,7-双苯-3-庚酮(DPHC,5-Hydroxy-1,7-diphenyl-3-heptanone)的LC-MS/MS方法,并应用于药动学研究。方法血浆样品经叔丁基甲醚萃取后进行分析,采用Kinetex XB-C_(18)色谱柱(2.10 mm×50mm,2.6μm),柱温40℃,以水(含0.01%甲酸)—甲醇(含0.01%甲酸)为流动相梯度洗脱,流速0.30 m L·min^(-1),ESI离子源,多反应离子监测(MRM),用于定量分析的离子对为m/z 283.3→265.1,内标化合物益智酮甲为313.1→137.0。结果 DPHC的线性范围为2.0~2 000.0 ng·mL^(-1)(r=0.995 7),最低定量限为2.0 ng·mL^(-1);提取回收率为75.62%~77.75%,基质效应为85.21%~90.24%;日内和日间精密度RSD均<15%。口服(po.)和静脉注射(iv.)DPHC主要药代动力学参数:C_(max)分别为(61.2±28.8),(246.7±6.7)ng·mL^(-1);t_(1/2)分别为(1.90±0.68),(0.34±0.20)h;AUC0-t分别为(100.4±33.8),(78.1±1.8)h·ng·mL^(-1)。结论本法经方法学验证,适用于大鼠血浆中DPHC浓度的测定,适合药动学研究。

OBJECTIVE To develop a rapid sensitive LC-MS/MS method for determination of 5-Hydroxy-1,7-diphenyl-3- heptanone（DPHC） in rat plasma, and apply the method in pharmacokinetics study furtherly. METHODS The plasma samples were extracted by methyl tert-butyl ether（MTBE）. The chromatographic analysis was carried out by Kinetex XB-C18 column （2.10 mm× 50 mm,2.6 μm）and the column temperature was 40 %. The mobile phase consisted of water（ contain 0.01% formic acid）and methanol （contain 0.01% formic acid） with gradient elution at a flow rate of 0.3 mL.min-I .Electrospray ionization（ESI） with a positive-ion and multiple reaction monitoring mode（MRM） was adopted.Yakucinone A was used as the internal standard, the precursor ion → product ion transitions m/z 283.3 →265.1 for DPHC and m/z 313.1 →137.0 for internal standard were monitored. RESULTS The plasma standard curve of DPHC were linear in the range of 2.0-2 000.0 ng· mL-1（ r = 0.995 7）.The limit of quantification was 2.0 ng· mL-1 .The plasma extraction recovery of DPHC was 75.62%- 77.75%.Matrix effect were 85.21%- 90.24%.The intra-day and inter-day RSDs were less than 15%. The main pharmaeokinetic parameters of po. and iv. were as follows ： Cmax were （ 61.2 ± 28.8 ） , （ 246.7± 6.7 ）ng· mL- 1 respectively ; t1/2 were（ 1.90± 0.68 ）, （ 0.34 ± 0.20 ） h respectively ; AUC0-t were （ 100.4 ± 33.8 ）, （ 78.1 ± 1.8 ） h· ng· mL-1 respectively. CONCLUSION The established method is proved to be suitable for determination of DPHC in rat plasma, it can be used as an effective method for the pharmacokineties studies.