Nara Akina, Saka Kanju, Yamada Chiho, Kodama Takanori, Takagi Tetsuya
1Division of Legal Medicine, Faculty of Medicine, Tohoku Medical and Pharmaceutical University, 1-15-1 Fukumuro, Miyagino-ku, Sendai-shi, Miyagi 983-8536 Japan.
2Department of Forensic Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033 Japan.
Forensic Toxicol. 2019;37(1):197-206. doi: 10.1007/s11419-018-0452-7. Epub 2018 Nov 19.
The potato glycoalkaloids (PGAs), α-solanine and α-chaconine can exert adverse effects on human health when consumed in excess. This study aimed to investigate the optimal extraction method for the quantitative analysis of PGAs in whole blood by using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and to apply this validated method to postmortem blood.
A total of 200 µL of human whole blood was prepared for PGA extraction. For validation, a solid-phase extraction (SPE) using Oasis PRiME HLB, in which extraction could be performed in three simple steps (sample loading, washing, and elution) was used, with no need for both conditioning and equilibration of columns for sample preparation.
In this method, the limit of detection and the lower limit of quantification (LLOQ) of both α-solanine and α-chaconine were 1 and 2 µg/L, respectively. The calibration curves of the two compounds were obtained with good linearity in the range of 2-100 µg/L. The recovery rates at the LLOQ of α-solanine and α-chaconine were ≥ 91.8% and ≥ 85.9%, respectively. The validation data (intra- and inter-day combined) for accuracy ranged from 93.5 to 106.6% for α-solanine and from 93.9 to 107.7% for α-chaconine. This validated method was successfully applied to one forensic autopsy case, and the concentrations of α-solanine and α-chaconine in the postmortem cardiac blood were 45.1 and 35.5 µg/L, respectively.
This validated UHPLC-MS/MS with SPE for quantitative analysis of PGAs could be useful in forensic toxicology.
马铃薯糖生物碱(PGAs),即α-茄碱和α-查茄碱,过量摄入时会对人体健康产生不利影响。本研究旨在探讨采用超高效液相色谱-串联质谱法(UHPLC-MS/MS)对全血中PGAs进行定量分析的最佳提取方法,并将该验证方法应用于死后血液。
制备200μL人全血用于PGA提取。为进行验证,使用了Oasis PRiME HLB固相萃取(SPE),其提取可通过三个简单步骤(样品加载、洗涤和洗脱)完成,无需对样品制备柱进行预处理和平衡。
在该方法中,α-茄碱和α-查茄碱的检测限和定量下限(LLOQ)分别为1μg/L和2μg/L。两种化合物的校准曲线在2-100μg/L范围内具有良好的线性。α-茄碱和α-查茄碱在LLOQ时的回收率分别≥91.8%和≥85.9%。α-茄碱的准确度验证数据(日内和日间合并)范围为93.5%至106.6%,α-查茄碱为93.9%至107.7%。该验证方法成功应用于一例法医尸检案例,死后心脏血液中α-茄碱和α-查茄碱的浓度分别为45.1μg/L和35.5μg/L。
这种经过验证的采用SPE的UHPLC-MS/MS法用于PGAs的定量分析,在法医毒理学中可能会有用。