Kwak Jae-Hwan, Kim Hye Kyung, Choe Sanggil, In Sangwhan, Pyo Jae Sung
College of Pharmacy, Kyungsung University, Busan 608-736, Republic of Korea.
Drug and Toxicology Section, Busan Institute, National Forensic Service, Yangsan 626-815, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Mar 15;1015-1016:209-213. doi: 10.1016/j.jchromb.2016.01.061. Epub 2016 Feb 22.
The main objective of this study was to develop and validate a simpler and less time consuming analytical method for determination of propofol glucuronide from hair sample, by using mixed mode anion exchange cartridge and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The study uses propofol glucuronide, a major metabolite of propofol, as a marker for propofol abuse. The hair sample was digested in sodium hydroxide solution and loaded in mixed-mode anion cartridge for solid phase extraction. Water and ethyl acetate were used as washing solvents to remove interfering substances from the hair sample. Consequently, 2% formic acid in ethyl acetate was employed to elute propofol glucuronide from the sorbent of mixed-mode anion cartridge, and analyzed by LC-MS/MS. The method validation parameters such as selectivity, specificity, LOD, LLOQ, accuracy, precision, recovery, and matrix effect were also tested. The linearity of calibration curves showed good correlation, with correlation coefficient 0.998. The LOD and LLOQ of the propofol glucuronide were 0.2 pg/mg and 0.5 pg/mg, respectively. The intra and inter-day precision and accuracy were acceptable within 15%. The mean values of recovery and matrix effect were in the range of 91.7-98.7% and 87.5-90.3%, respectively, signifying that the sample preparation, washing and extraction procedure were efficient, and there was low significant hair matrix effect for the extraction of propofol glucuronide from hair sample on the mixed mode anion cartridge. To evaluate the suitability of method, the hair of propofol administered rat was successfully analyzed with this method.
本研究的主要目的是开发并验证一种更简单、耗时更少的分析方法,用于通过混合模式阴离子交换柱和液相色谱-串联质谱法(LC-MS/MS)从毛发样本中测定丙泊酚葡萄糖醛酸苷。该研究使用丙泊酚的主要代谢物丙泊酚葡萄糖醛酸苷作为丙泊酚滥用的标志物。将毛发样本在氢氧化钠溶液中消化,然后加载到混合模式阴离子柱上进行固相萃取。用水和乙酸乙酯作为洗涤溶剂,以去除毛发样本中的干扰物质。随后,使用乙酸乙酯中的2%甲酸从混合模式阴离子柱的吸附剂上洗脱丙泊酚葡萄糖醛酸苷,并通过LC-MS/MS进行分析。还测试了方法验证参数,如选择性、特异性、检测限(LOD)、定量下限(LLOQ)、准确度、精密度、回收率和基质效应。校准曲线的线性显示出良好的相关性,相关系数为0.998。丙泊酚葡萄糖醛酸苷的LOD和LLOQ分别为0.2 pg/mg和0.5 pg/mg。日内和日间精密度和准确度在15%以内是可接受的。回收率和基质效应的平均值分别在91.7-98.7%和87.5-90.3%范围内,这表明样品制备、洗涤和萃取程序是有效的,并且在混合模式阴离子柱上从毛发样本中提取丙泊酚葡萄糖醛酸苷时,毛发基质效应较低且不显著。为了评估该方法的适用性,用该方法成功分析了给予丙泊酚的大鼠的毛发。