【摘要】： 目的 本文建立了血漿中滅多威的固相萃取-高效液相色譜(SPE-HPLC)快速定性定量分析方法,并利用固相萃取-高效液相色譜-質(zhì)譜(SPE-HPLC-MS)法對(duì)大鼠血漿中的滅多威及其代謝物進(jìn)行了研究。 方法 以空白大鼠血漿添加滅多威標(biāo)準(zhǔn)品及內(nèi)標(biāo)物安定標(biāo)準(zhǔn)品對(duì)血漿樣品的前處理方法、儀器測(cè)試條件、特異性、回收率、靈敏度、精密度、準(zhǔn)確度、線性關(guān)系、穩(wěn)定性進(jìn)行全面考察,建立血漿中滅多威的SPE-HPLC快速定性定量分析方法;HPLC-MS法在大鼠血漿中檢出了滅多威及其代謝物。 采用HPLC法:測(cè)定不同時(shí)間點(diǎn)大鼠血漿中滅多威的藥物濃度。按照滅多威5mg/kg給雄性大鼠灌胃后,收集0.33h,0.67h,1h,2h,3h,5h,7h,9h,11h,13h,15h,17h,19h,21h,23h的大鼠血漿,SPE-HPLC法測(cè)定血漿中滅多威,并對(duì)24h內(nèi)大鼠血漿中滅多威濃度的變化規(guī)律進(jìn)行了描述;模擬法醫(yī)鑒定,按照滅多威50mg/kg給雄性大鼠灌胃后,將大鼠尸體室溫存放72h,取心血,SPE-HPLC法檢測(cè)大鼠血漿中滅多威的含量;色譜柱:kromasil C_(18)柱(4.6mm i.d.×250mm,5μm粒徑);流動(dòng)相:甲醇:水=40:60(V/V);檢測(cè)波長(zhǎng):235nm;安定作為內(nèi)標(biāo)。 采用HPLC-MS法:分析大鼠血漿中滅多威及其代謝物。模擬法醫(yī)鑒定,按照滅多威50mg/kg給雄性大鼠灌胃后,將大鼠尸體室溫存放72h,取心血,SPE-HPLC-MS法檢測(cè)大鼠血漿中的滅多威及其代謝物。色譜柱:Xterra C_(18)柱(2.1mm i.d.×150mm,5μm粒徑);流動(dòng)相:甲醇:水=15:85(V/V);檢測(cè)波長(zhǎng):235nm;質(zhì)譜:采用電噴霧電離方式(ESI+),毛細(xì)管電壓:3.0kV;離子源溫度:110℃;干燥氣溫度:300℃;干燥氣流速:450L/h;掃描范圍:50～300m/z。 結(jié)果 SPE-HPLC法測(cè)得滅多威在血漿中的線性范圍是0.1μg/ml～20μg/ml(r=0.9993,P0.001),血漿中的檢測(cè)限是0.03μg/ml(S/N=3),日內(nèi)、日間精密度的RSD值分別在8.33%與11.11%以?xún)?nèi),日內(nèi)、日間準(zhǔn)確度值分別在90%與120%之間,回收率值在88%±4.4%以上;并且在灌服大量的滅多威引起中毒死亡72h后的大鼠血漿中仍能檢測(cè)到滅多威。 SPE-HPLC-MS法發(fā)現(xiàn),給藥后的大鼠血漿中,除滅多威原體外,還有一種代謝物,并測(cè)得其準(zhǔn)分子離子峰及其各級(jí)碎片離子,經(jīng)與對(duì)照品比較及質(zhì)譜斷裂規(guī)律推斷滅多威在大鼠血漿中的代謝產(chǎn)物為S-甲基-N-羥基硫代乙酰亞胺酯。 結(jié)論 所建立的分析方法可靠、實(shí)用、便捷,可對(duì)血漿中的滅多威進(jìn)行定性定量地快速測(cè)定,并且可檢出其代謝物,為滅多威中毒的法醫(yī)鑒定及臨床檢驗(yàn)進(jìn)一步提供了參考。
[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，

本文編號(hào)：2120042