大鼠血浆中灭多威的分析及其代谢物的质谱研究
发布时间：2018-07-13 16:29
【摘要】： 目的 本文建立了血浆中灭多威的固相萃取-高效液相色谱(SPE-HPLC)快速定性定量分析方法,并利用固相萃取-高效液相色谱-质谱(SPE-HPLC-MS)法对大鼠血浆中的灭多威及其代谢物进行了研究。 方法 以空白大鼠血浆添加灭多威标准品及内标物安定标准品对血浆样品的前处理方法、仪器测试条件、特异性、回收率、灵敏度、精密度、准确度、线性关系、稳定性进行全面考察,建立血浆中灭多威的SPE-HPLC快速定性定量分析方法;HPLC-MS法在大鼠血浆中检出了灭多威及其代谢物。 采用HPLC法:测定不同时间点大鼠血浆中灭多威的药物浓度。按照灭多威5mg/kg给雄性大鼠灌胃后,收集0.33h,0.67h,1h,2h,3h,5h,7h,9h,11h,13h,15h,17h,19h,21h,23h的大鼠血浆,SPE-HPLC法测定血浆中灭多威,并对24h内大鼠血浆中灭多威浓度的变化规律进行了描述;模拟法医鉴定,按照灭多威50mg/kg给雄性大鼠灌胃后,将大鼠尸体室温存放72h,取心血,SPE-HPLC法检测大鼠血浆中灭多威的含量;色谱柱:kromasil C_(18)柱(4.6mm i.d.×250mm,5μm粒径);流动相:甲醇:水=40:60(V/V);检测波长:235nm;安定作为内标。 采用HPLC-MS法:分析大鼠血浆中灭多威及其代谢物。模拟法医鉴定,按照灭多威50mg/kg给雄性大鼠灌胃后,将大鼠尸体室温存放72h,取心血,SPE-HPLC-MS法检测大鼠血浆中的灭多威及其代谢物。色谱柱:Xterra C_(18)柱(2.1mm i.d.×150mm,5μm粒径);流动相:甲醇:水=15:85(V/V);检测波长:235nm;质谱:采用电喷雾电离方式(ESI+),毛细管电压:3.0kV;离子源温度:110℃;干燥气温度:300℃;干燥气流速:450L/h;扫描范围:50～300m/z。 结果 SPE-HPLC法测得灭多威在血浆中的线性范围是0.1μg/ml～20μg/ml(r=0.9993,P0.001),血浆中的检测限是0.03μg/ml(S/N=3),日内、日间精密度的RSD值分别在8.33%与11.11%以内,日内、日间准确度值分别在90%与120%之间,回收率值在88%±4.4%以上;并且在灌服大量的灭多威引起中毒死亡72h后的大鼠血浆中仍能检测到灭多威。 SPE-HPLC-MS法发现,给药后的大鼠血浆中,除灭多威原体外,还有一种代谢物,并测得其准分子离子峰及其各级碎片离子,经与对照品比较及质谱断裂规律推断灭多威在大鼠血浆中的代谢产物为S-甲基-N-羟基硫代乙酰亚胺酯。 结论 所建立的分析方法可靠、实用、便捷,可对血浆中的灭多威进行定性定量地快速测定,并且可检出其代谢物,为灭多威中毒的法医鉴定及临床检验进一步提供了参考。
[Abstract]:Objective to establish a rapid qualitative and quantitative analysis method for methomyl in plasma by solid phase extraction and high performance liquid chromatography (SPE-HPLC). Solid phase extraction-high performance liquid chromatography-mass spectrometry (SPE-HPLC-MS) was used to study methomyl and its metabolites in rat plasma. Methods Plasma samples were pretreated by adding methomyl standard and diazepam in blank rat plasma, the instrument test conditions, specificity, recovery rate, sensitivity, precision and accuracy. The linear relationship and stability were investigated. A SPE-HPLC rapid qualitative and quantitative analysis method was established for the determination of methomyl and its metabolites in rat plasma by HPLC-MS. HPLC method: to determine the concentration of methomyl in rat plasma at different time points. After intragastric administration of medovir 5mg/kg in male rats, we collected 0.33 h 0.67 h ~ 1 h ~ 2 h ~ 3 h ~ 5 h ~ 7 h ~ 9 h ~ 1 ~ 11 h ~ (13) h ~ (15) h ~ (17) h ~ (19) h ~ (21) h ~ (23) h ~ (23 h) for the determination of methomyl in plasma, and described the change rule of the concentration of methomyl in rat plasma within 24 h. The rat cadavers were stored at room temperature for 72 hours according to methomyl 50mg/kg. The content of methomyl in rat plasma was determined by SPE-HPLC. The column was 4.6mm i.d. 脳 250mm-1 (5 渭 m). The mobile phase was methanol: water 40: 60 (V / V); the detection wavelength was: 235nm; diazepam was used as internal standard. The method of HPLC-MS was used to analyze methomyl and its metabolites in rat plasma. After the male rats were fed with methomyl 50mg/kg at room temperature for 72 hours, the blood samples were collected for the determination of methomyl and its metabolites in the plasma of rats by SPE-HPLC-MS. The mobile phase consisted of methanol: water 15: 85 (V / V); detection wavelength: 1: 235 nm; mass spectrometry: electrospray ionization (ESI), capillary voltage: 3.0 kV; ion source temperature: 110 鈩，
本文编号：2120042
[Abstract]:Objective to establish a rapid qualitative and quantitative analysis method for methomyl in plasma by solid phase extraction and high performance liquid chromatography (SPE-HPLC). Solid phase extraction-high performance liquid chromatography-mass spectrometry (SPE-HPLC-MS) was used to study methomyl and its metabolites in rat plasma. Methods Plasma samples were pretreated by adding methomyl standard and diazepam in blank rat plasma, the instrument test conditions, specificity, recovery rate, sensitivity, precision and accuracy. The linear relationship and stability were investigated. A SPE-HPLC rapid qualitative and quantitative analysis method was established for the determination of methomyl and its metabolites in rat plasma by HPLC-MS. HPLC method: to determine the concentration of methomyl in rat plasma at different time points. After intragastric administration of medovir 5mg/kg in male rats, we collected 0.33 h 0.67 h ~ 1 h ~ 2 h ~ 3 h ~ 5 h ~ 7 h ~ 9 h ~ 1 ~ 11 h ~ (13) h ~ (15) h ~ (17) h ~ (19) h ~ (21) h ~ (23) h ~ (23 h) for the determination of methomyl in plasma, and described the change rule of the concentration of methomyl in rat plasma within 24 h. The rat cadavers were stored at room temperature for 72 hours according to methomyl 50mg/kg. The content of methomyl in rat plasma was determined by SPE-HPLC. The column was 4.6mm i.d. 脳 250mm-1 (5 渭 m). The mobile phase was methanol: water 40: 60 (V / V); the detection wavelength was: 235nm; diazepam was used as internal standard. The method of HPLC-MS was used to analyze methomyl and its metabolites in rat plasma. After the male rats were fed with methomyl 50mg/kg at room temperature for 72 hours, the blood samples were collected for the determination of methomyl and its metabolites in the plasma of rats by SPE-HPLC-MS. The mobile phase consisted of methanol: water 15: 85 (V / V); detection wavelength: 1: 235 nm; mass spectrometry: electrospray ionization (ESI), capillary voltage: 3.0 kV; ion source temperature: 110 鈩，
本文编号：2120042
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