Tsuchihashi H, Nishikawa M, Igarashi K, Tatsuno M, Katagi M, Kasuya F, Fukui M
Forensic Science Laboratory, Osaka Prefectural Police Headquarters, Japan.
J Anal Toxicol. 1998 Nov-Dec;22(7):591-5. doi: 10.1093/jat/22.7.591.
A simple, rapid, and sensitive method which allows us to simultaneously determine bromvalerylurea (BVU) and its three metabolites (3-methylbutyrylurea [MVU], alpha-(cystein-S-yl)isovalerylurea [CVU], and alpha-(N-acetylcystein-S-yl)isovalerylurea [AcCVU]) was investigated by frit-fast atom bombardment liquid chromatography-mass spectrometry (frit-FAB LC-MS). The LC-MS analysis was performed after the solid-phase extraction from tissue and urine samples with a Sep-Pak C18 cartridge. Tissue homogenates and urine were adjusted to pH 4.0 and applied to the cartridges. The retained BVU and its metabolites were eluted from the cartridge with 2 mL of acetonitrile/10 mM ammonium acetate buffer (pH 3.5, 50:50, v/v). The eluate was analyzed by LC-MS, which employs a semimicro type L-column ODS column. The proposed conditions are as follows: mobile phase A, 0.4% glycerol in acetonitrile/10 mM ammonium acetate buffer (pH 3.5) (5:95, v/v); mobile phase B, 0.4% glycerol in acetonitrile; elution mode, linear gradient, 100% A (5 min) to 100% B in 15 min; flow rate, 0.2 mL/min; split ratio, 1:40. Extraction recoveries of BVU and its metabolites were 91.90-97.79% from the spiked liver homogenate and 89.68-96.13% from the spiked urine. The detection limits ranged from 10 to 25 ng/g in selected ion monitoring mode.
通过多孔板快速原子轰击液相色谱 - 质谱联用仪（frit - FAB LC - MS）研究了一种简单、快速且灵敏的方法，该方法可让我们同时测定溴戊酰脲（BVU）及其三种代谢物（3 - 甲基丁酰脲[MVU]、α -（半胱氨酸 - S - 基）异戊酰脲[CVU]和α -（N - 乙酰半胱氨酸 - S - 基）异戊酰脲[AcCVU]）。在用Sep - Pak C18柱对组织和尿液样本进行固相萃取后进行LC - MS分析。将组织匀浆和尿液调节至pH 4.0后应用于柱子。保留的BVU及其代谢物用2 mL乙腈/10 mM醋酸铵缓冲液（pH 3.5，50:50，v/v）从柱子上洗脱下来。洗脱液通过LC - MS进行分析，该LC - MS采用半微量型L - 柱ODS柱。建议的条件如下：流动相A，乙腈/10 mM醋酸铵缓冲液（pH 3.5）（5:95，v/v）中含0.4%甘油；流动相B，乙腈中含0.4%甘油；洗脱模式，线性梯度，100%A（5分钟）至15分钟内变为100%B；流速，0.2 mL/分钟；分流比，1:40。从加标的肝脏匀浆中提取BVU及其代谢物的回收率为91.90 - 97.79%，从加标的尿液中提取的回收率为89.68 - 96.13%。在选择离子监测模式下，检测限为10至25 ng/g。
