在线固相萃取-液质联用法定量分析生物基质中丁香醛及其在大鼠体内药代动力学研究中的应用

The development and validation of an on-line solid phase extraction HPLC-MS/MS method for quantification of syringaldehyde in biological matrix and its application in Sprague-Dawley rat pharmacokinetic study

目的建立测定SD大鼠血浆中丁香醛的在线固相萃取-液质联用法(on-line SPE LC-MS/MS)。方法采用乙腈沉淀蛋白后,取上清经on-line SPE系统进行血浆样品预处理,选用香兰素为内标,以LC-MS/MS法测定大鼠血浆中丁香醛的含量。色谱柱为GRACE Alltima HP C18(50 mm×2.1 mm,5μm),流动相为乙腈-0.1%甲酸(60∶40,V∶V),流速为0.4 ml·min-1。电喷雾离子化(ESI)方式,采用多反应监测,检测离子为正离子,分别选择m/z 182.3→m/z123.1和m/z 153.1→m/z 93.0作为丁香醛和内标物香兰素的检测离子对。结果丁香醛在10.0～2 000μg·L-1范围内线性关系良好(r=0.9997)。方法的准确度(相对误差,RE)范围为-8.5%～5.9%;日内精密度(相对标准偏差,RSD)<16.3%,日间精密度RSD<13.8%。SD大鼠灌胃给予丁香醛(17.5 mg.kg-1和70 mg.kg-1)后,主要药动学参数:T12分别为41.9 min和54.0 min、Cmax分别为361.0μg·L-1和944.0μg·L-1、Tmax分别为10.0 min和27.5 min、CL分别为848.1 ml·min-1.kg-1和683.9 ml·min-1·kg-1。结论建立的基质中丁香醛的测定方法快速、准确、特异性好,适用于丁香醛在SD大鼠体内的药代动力学研究。

Aim To establish an on-line solid phase extraction high performance liquid chromatography tandem mass spectrometric（on-line SPE LC-MS/MS） method for quantification of syringaldehyde in Sprague-Dawley rat plasma.Methods After precipitation with acetonitrile,the supernatant was separated by on-line SPE system on an extract column.Vanillin was used as internal standard.Syringaldehyde in Sprague-Dawley rat plasma was determined by LC-MS/MS.GRACE Alltima HP C18 column（50 mm×2.1 mm,5 μm） was used as analytical column.The mobile phase consisted of acetonitrile and 0.1 % formic acid（60 ∶40,V ∶V）,which was pumped at flow rate with 0.4 ml·min-1.The analyte was detected using electro-spray ionization（ESI） interface with positive ionization mode.Ions were monitored in multiple reaction monitoring（MRM） modes.The fragmentation transitions were m/z 182.3→m/z 123.1 for syringaldehyde and m/z 153.1→m/z 93.0 for vanillin.Results The linear range（r=0.9997） of the present method was 10.0~2 000 μg·L-1.The accuracy（relative error,RE） of method ranged from-8.5% to 5.9%.The intra-day and inter-day precision（Relative Standard Deviation,RSD） were within 16.3% and 13.8%,respectively.After intragastric administration（at dosage of 17.5 mg·kg-1 and 70 mg·kg-1） to Sprague-Dawley rats,the main pharmacokinetic parameters were as follows： the elimination half life were 41.9 min and 54.0 min,the peak plasma concentration were 361.0 μg·L-1 and 944.0 μg·L-1,the time of arriving to peak plasma concentration were 10.0 min and 27.5 min,the clearance were 848.1 ml·min-1·kg-1 and 683.9 ml·min-1·kg-1,respectively.Conclusion This newly developed method is accuracy,precision and of high throughput,which could be applied to pharmacokinetic study of syringaldehyde in Sprague-Dawley rats.