Cho Byungsuk, Kim Suncheun, In Sangwhan, Choe Sanggil
National Forensic Service, 10 Ipchun-ro, Wonju, Gangwon-do 26460, South Korea.
National Forensic Service, 10 Ipchun-ro, Wonju, Gangwon-do 26460, South Korea.
Forensic Sci Int. 2017 Sep;278:304-312. doi: 10.1016/j.forsciint.2017.07.024. Epub 2017 Jul 29.
A liquid chromatography-tandem mass spectrometry method with solid-phase extraction (SPE) was developed and validated for the detection and quantitation of bentazone and its two hydroxylated metabolites, 6-hydroxybentazone and 8-hydroxybentazone, in postmortem blood. Sample cleanup was performed using a hydrophilic-lipophilic balanced (HLB) SPE cartridge and then separated on a C18 LC column using a gradient elution of 0.1% formic acid in distilled water and 0.1% formic acid in methanol. The identification of bentazone and its hydroxylated metabolites was performed using tandem mass spectrometry with electrospray ionization in negative ion mode with selective reaction monitoring. The retention times of bentazone, 6-hydroxybentazone, 8-hydroxybentazone, and 2-methyl-4-chlorophenoxyacetic acid (MCPA, internal standard) appeared separately in the chromatogram. The matrix effect, recovery, and process efficiency of bentazone were 75.3%, 103.6% and 77.9%, respectively. In addition, good accuracy (88.2-110.5%), precision (0.5-7.5%, bias), and linearity (5-500ng/mL) were obtained with this method. The limit of detection (LOD) of bentazone, 6-hydroxybentazone, and 8-hydroxybentazone were 0.05, 0.5, and 0.5ng/mL, respectively. The method developed herein was applied to authentic samples from three fatal cases from 2016 for the determination of the corresponding bentazone and its metabolites levels. The concentration ranges of bentazone, 6-hydroxybentazone, and 8-hydroxybentazone in the heart blood from the three victims were 46.0-91.8, 4.2-6.2, and 0.2-0.6μg/mL, respectively.
建立了一种采用固相萃取（SPE）的液相色谱-串联质谱法，用于检测和定量死后血液中苯达松及其两种羟基化代谢物6-羟基苯达松和8-羟基苯达松。使用亲水亲脂平衡（HLB）固相萃取柱进行样品净化，然后在C18液相色谱柱上进行分离，采用0.1%甲酸水溶液和0.1%甲酸甲醇溶液进行梯度洗脱。采用电喷雾电离负离子模式下的串联质谱和选择性反应监测对苯达松及其羟基化代谢物进行鉴定。苯达松、6-羟基苯达松、8-羟基苯达松和2-甲基-4-氯苯氧基乙酸（MCPA，内标）的保留时间在色谱图中分别出现。苯达松的基质效应、回收率和处理效率分别为75.3%、103.6%和77.9%。此外，该方法具有良好的准确度（88.2-110.5%）、精密度（0.5-7.5%，偏差）和线性（5-500ng/mL）。苯达松、6-羟基苯达松和8-羟基苯达松的检测限（LOD）分别为0.05、0.5和0.5ng/mL。本文建立的方法应用于2016年三例死亡病例真实样本中相应苯达松及其代谢物水平的测定。三名受害者心血中苯达松、6-羟基苯达松和8-羟基苯达松的浓度范围分别为46.0-91.8、4.2-6.2和0.2-0.6μg/mL。
