Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China; Modern Research Center for Traditional Chinese Medicine, Second Military Medical University, Shanghai 200433, China.
Modern Research Center for Traditional Chinese Medicine, Second Military Medical University, Shanghai 200433, China; Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
Chin J Nat Med. 2014 Feb;12(2):154-60. doi: 10.1016/S1875-5364(14)60026-1.
To develop an HPLC-MS/MS method for the quantification of platycodin D (PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract (PRE) containing PD.
Plasma samples were pretreated with solid-phase extraction using an Oasis® HLB SPE cartridge. Madecassoside was used as the internal standard (IS). Chromatographic separation was achieved on an ODS column (100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water (30 : 70, V/V) containing 0.1 mmol·L(-1) ammonium acetate at a flow rate of 0.25 mL·min(-1). The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization (ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside (IS), respectively.
The calibration curve was linear from 5 to 2 000 ng·mL(-1) (r(2) >0.99) with a lower limit of quantification (LLOQ) of 5 ng·mL(-1). The intra- and inter-day precision (relative standard deviation, RSD) values were below 15% and the accuracy (relative error, RE) was from -15% to +15% at three quality control (QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be (0.48 ± 0.19)% when administered PD, and to be (1.81 ± 0.89)% when administered PRE.
The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.
建立一种 HPLC-MS/MS 法测定大鼠血浆中桔梗皂苷 D（PD）的含量，并获取口服纯 PD 或含 PD 的桔梗提取物（PRE）后的 PD 主要药代动力学参数。
采用 Oasis®HLB SPE 小柱固相萃取法预处理血浆样品，以玛咖皂苷元为内标（IS）。色谱分离在 ODS 柱（100 mm×2.1 mm i.d.，3.5 μm）上进行，流动相为乙腈/水（30：70，V/V），含 0.1 mmol·L（-1）乙酸铵，流速为 0.25 mL·min（-1）。采用电喷雾电离（ESI）源，三重四极串联质谱进行检测，色谱运行时间为 3.0 min。检测时，PD 的监测离子对为 m/z 1 223.6→469.2，IS 为 m/z 973.6→469.2，采用多重反应监测（MRM）模式进行检测。
PD 的校准曲线在 52 000 ng·mL（-1）范围内线性良好（r（2）>0.99），定量下限（LLOQ）为 5 ng·mL（-1）。三个质控（QC）浓度水平的日内和日间精密度（相对标准差，RSD）均低于 15%，准确度（相对误差，RE）在-15%+15%范围内。分别静脉注射 PD 和口服 PD、PRE 后 24 h 测定大鼠血浆中 PD 浓度。发现大鼠口服 PD 的绝对生物利用度为（0.48±0.19）%，口服 PRE 的生物利用度为（1.81±0.89）%。
该 HPLC-MS/MS 法可用于评价 PD 及桔梗提取物在大鼠体内的药代动力学参数和口服生物利用度。
