Wen Qi, Li Hai-Long, Chen Feng, Gong Jing-Wen, Guan Wei-Wei, Fu Nai-Guang, Tan Yin-Feng
Department of Pharmacy, Hainan Medical College, Haikou, China.
Department of Pharmacy, Hainan Medical College, Hainan Provincial Key Laboratory of R&D on Tropical Herbs, Haikou, China.
Biomed Chromatogr. 2018 Dec;32(12):e4373. doi: 10.1002/bmc.4373. Epub 2018 Sep 26.
The aim of the present study was to develop a rapid, specific and sensitive LC-MS/MS method for the determination of DPHB [7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-4-hepten-3-one] in rat plasma using yakuchinone A as an internal standard (IS). n-Hexane was used for the extraction of DPHB from rat plasma. Chromatographic separation of DPHB was achieved using a Kinetex XB-C column (2.10 × 50 mm, 2.6 μm) at 40°C. The mobile phase consisted of water (containing 0.1‰ formic acid, A) and acetonitrile (containing 0.1‰ formic acid, B) under a gradient elution at a flow rate of 0.3 mL min . Positive electrospray ionization and multiple reaction monitoring mode were used for detection. The selected precursor ion to product ion pairs, m/z 311.3 → 137.0 for DPHB and m/z 313.1 → 137.0 for yakuchinone A, were monitored. Good linearity was observed over the concentration range from 2 to 2000 ng mL (r = 0.9969). The recovery efficiency of DPHB from rat plasma was 54.8-69.7%, while the matrix effect ranged from 99.7 to 113%. Intra- and inter-day precision and accuracy values were within ±15% at three different quality control concentration levels. This validated method was successfully applied to pharmacokinetic studies in rats after a single p.o. or i.v. dose of DPHB solution. The route of administration significantly influenced systemic exposure to DPHB, and low bioavailability of DPHB was observed. The method developed here will be further improved and used in future pharmacokinetic studies.
本研究的目的是开发一种快速、特异且灵敏的液相色谱-串联质谱（LC-MS/MS）方法，以犛牛儿酮A作为内标物（IS）来测定大鼠血浆中的DPHB [7-(4″-羟基-3″-甲氧基苯基)-1-苯基-4-庚烯-3-酮]。正己烷用于从大鼠血浆中提取DPHB。使用Kinetex XB-C柱（2.10×50 mm，2.6μm）在40°C下实现DPHB的色谱分离。流动相由水（含0.1‰甲酸，A）和乙腈（含0.1‰甲酸，B）组成，采用梯度洗脱，流速为0.3 mL/min。采用正电喷雾电离和多反应监测模式进行检测。监测选定的前体离子到产物离子对，DPHB为m/z 311.3→137.0，犛牛儿酮A为m/z 313.1→137.0。在2至2000 ng/mL的浓度范围内观察到良好的线性关系（r = 0.9969）。DPHB从大鼠血浆中的回收率为54.8 - 69.7%，而基质效应范围为99.7至113%。在三个不同的质量控制浓度水平下，日内和日间精密度及准确度值均在±15%以内。该验证方法成功应用于大鼠单次口服或静脉注射DPHB溶液后的药代动力学研究。给药途径显著影响DPHB的全身暴露，且观察到DPHB的生物利用度较低。此处开发的方法将进一步改进并用于未来的药代动力学研究。
