目的 采用液相色譜-質(zhì)譜聯(lián)用法(LC-MS/MS)測定人血漿中烏苯美司的藥物濃度,并應(yīng)用于健康受試者體內(nèi)藥動學(xué)研究。方法 以文拉法辛為內(nèi)標(biāo),血漿樣品經(jīng)甲醇沉淀蛋白后,以甲醇-醋酸銨(2 mmol·L^-1,含 0.2% 甲酸)水溶液(50:50)為流動相,Waters 公司 XTerra^® MS C₁₈(150 mm×4.6 mm,5 μm)色譜柱分離,體積流量為 900 μL·min^-1,每個樣品的分析時間為 4.20 min。樣品經(jīng)電噴霧離子源正離子化后,通過三重四極桿串聯(lián)質(zhì)譜儀,在多反應(yīng)監(jiān)測模式下測定烏苯美司(m/z 309.3→120.2)和內(nèi)標(biāo)文拉法辛(m/z 278.2→215.1)的濃度。20 名受試者空腹口服烏苯美司膠囊 20 mg,250 mL溫開水送服,于用藥前和用藥后10、20、30、45 min及1.0、1.5、2.0、3.0、4.0、6.0、8.0、10.0 h由肘靜脈取血3 mL,制備血漿,按建立的LC-MS/MS法測定血漿中烏苯美司濃度,計(jì)算藥動學(xué)參數(shù)。結(jié)果 烏苯美司的血漿質(zhì)量濃度在1.0~2 500.0 ng·L^-1范圍內(nèi)線性關(guān)系良好,定量下限為 1.000 ng·mL^-1,批內(nèi)、批間精密度(RSD)均在 2.2%~5.1%,相對偏差(RE)在±15% 范圍內(nèi)。烏苯美司血漿樣品室溫放置 24 h,反復(fù)凍融(-20℃)3 次及冰凍(-20℃)保存 50 d 的情況下均穩(wěn)定。健康受試者空腹口服烏苯美司膠囊20mg后,血漿中烏苯美司的達(dá)峰濃度(C_max)為(1 375±298) ng·mL^-1, 藥時曲線下面積(AUC_{0~10 h})為(2 106±296) h·ng·mL^-1,AUC_0~∞為(2 116±299) h·ng·mL^-1,半衰期(t_1/2)為(1.38±0.20)h,達(dá)峰時間(t_max)為(0.71±0.23) h。結(jié)論 建立的LC-MS/MS分析方法簡便、選擇性高、靈敏度高,可用于受試者空腹口服 20 mg 烏苯美司膠囊后血漿樣品中烏苯美司的藥動學(xué)研究,烏苯美司膠囊口服吸收迅速,0.71 h 后血藥濃度可達(dá)峰值。

Objective To establish an LC-MS/MS method for the determination of ubenimex, which was used subsequently to investigate the pharmacokinetics of ubenimex in healthy Chinese volunteers. Methods Venlafaxine was used as internal standard. Following a deproteinization procedure, ubenimex and the internal standard venlafaxine were eluted isocratically using a mobile phase containing of methanol-2 mmol·L^-1 ammonium acetate and 0.2% formic acid aqueous solution (50: 50) at a flow rate of 900 μL·min^-1 within 4.20 min. The analysis was carried out by using Waters XTerra^® MS C₁₈ column (150 mm × 4.6 mm, 5 μm). Ubenimex and the internal standard were measured by a triple-quadrupole mass spectrometer in positive electron electronic spray ion (ESI) mode using multiple reaction monitoring (MRM). The extracted ions monitored following MRM transitions were m/z 309.3→ 120.2 for ubenimex and m/z 278.2→215.1 for the internal standard venlafaxine. Twenty subjects took orally 20 mg of Ubenmex Capsule on an fasted state, 250 mL of warm boiled water, and took 3 mL of blood from the elbow vein before administration and 10, 20, 30, 45 minutes and 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 10.0 h after administration, to prepare plasma. The concentration of ubenmex in plasma was determined by the established LC-MS/MS method, and the pharmacokinetic parameters were calculated. Results The calibration curve of ubenimex in human plasma was linear over the concentration rang of (1.0-2 500.0) ng·mL^-1. The lower limit of quantitation was 1.000 ng·mL^-1. The intra-and inter-run precisions at three quality control levels were within 2.2%-5.1%, the relative deviation of the assay was within -15.0%-15.0%. The plasma samples were stable at room temperature for 24 h, at -20℃ for 50 days and during three freeze-thaw cycles. After the healthy subjects took Ubenimex Capsule 20 mg on an fasted state, the main pharmacokinetic parameters of ubenimex were as follows: C_max was (1 375 ± 298) ng·mL^-1, AUC_0-10h was (2 106 ± 296) h·ng·mL^-1, AUC_0-∞ was (2 116 ± 299) ng·h·mL^-1, t_1/2 was (1.38 ± 0.20) h, t_max was (0.71 ± 0.23) h. Conclusion The method was proved to be convenient, accurate and sensitive. The method was proved to be suitable for the pharmacokinetics of ubenimex in healthy Chinese volunteers after a single oral dose of 20 mg Ubenimex Capsule. Ubenimex Capsule were rapidly absorbed by oral administration, and the blood drug concentration reached the peak after 0.71 hours.

R285.61

蘇州市藥學(xué)會-江蘇恒瑞臨床藥學(xué)科研基金項(xiàng)目(Syhky201807)