畜產品中動物用藥Albendazole、Thiabendazole、Mebendazole及其代謝產物檢驗方法之建立   全文下載

Simultaneous Determination of Albendazole, Thiabendazole, Mebendazole and Their Metabolites in Livestock by High Performance Liquid Chromatography

本研究建立以高效液相層析法（high performance liquid chromatography, HPLC）同時檢測畜產品中thiabendazole、5-hydroxythiabendazole、albendazole sulfoxide、albendazole sulfone、albendazole 2-aminosulfone及mebendazole等六種benzimidazoles之分析方法。檢體中之benzimidazoles在鹼性情況下以乙?萃取，經Sep-Pak C18萃取匣淨化後，再以HPLC分析。結果發現利用Cosmosil 5C18 MS-II（5μm, 4.6mm i.d. × 250mm）層析管柱，以乙?與0.02M磷酸二氫鈉溶液（pH3.3）為梯度移動相溶液，於光二極體列陣檢出器及螢光檢出器偵測下，benzimidazoles之分離效果及重複性皆佳。經以0.05~2.50μg／mL之六種benzimidazoles混合標準溶液所製作標準曲線之相關係數皆大於0.999。檢體中添加相當於0.02、0.10及1.00ppm六種benzimidazoles之平均回收率，除了低濃度之5-hydroxythiabendazole及mebendazole介於71.4~87.1%外，其餘回收率介於81.6~104.3%之間，且變異係數皆小於5.9%。本方法之最低檢出限量，紫外光檢測介於0.005~0.030 ppm，螢光檢測介於0.004~0.020 ppm，同日內及異日間之變異係數分別小於1.92%及7.22%，顯示本方法之精密度可接受。進而利用本方法分析市售豬肉、豬肝、牛肉、羊肉、牛奶及羊奶各5件，結果均未檢出benzimidazoles。"

An analytical method using high performance liquid chromatography (HPLC) was developed for the simultaneous determination of thiabendazole, 5-hydroxythiabendazole, albendazole sulfoxide, albendazole sulfone, albendazole 2-aminosulfone and mebendazole in livestock. These six benzimidazoles were extracted with acetonitrile under basic conditions, followed by a Sep-Pak C18 cartridge cleaning procedure. The HPLC separation was achieved on a Cosmosil 5C18-MS-II column (5 µm, 4.6 mm i.d. × 250 mm) using gradient elution with acetonitrile and 0.02M sodium dihydrogen phosphate (pH 3.3), and detection was performed with photodiode array and fluorescence detector. Good linearity was observed from the calibration plot at concentrations from 0.05 to 2.50 µg/mL. Recovery studies of the analytes were performed at 0.02, 0.10 and 1.00 ppm spiked levels. The average recovery of 5-hydroxythiabendazole and mebendazole at low concentrations ranged from 71.4 to 87.1% and ranged from 81.6 to 104.3% at other concentrations, with coefficient of variation less than 5.9%. The detection limit for benzimidazoles was between 0.005~0.030 ppm under photodiode array detection and 0.004~0.020 ppm under fluorescence detection. The coefficient of variation of intra-day and inter-day assays was lower than 1.92% and 7.22%, respectively. These results indicated that the developed method had an acceptable precision. This method was applied to detect benzimidazoles in samples of pork, swine liver, beef, lamb, bovine milk and goat milk purchased from various markets in Taipei. The results showed that none of the six benzimidazoles were detected in the thirty samples. 本研究建立以高效液相層析法（high performance liquid chromatography, HPLC）同時檢測畜產品中thiabendazole、5-hydroxythiabendazole、albendazole sulfoxide、albendazole sulfone、albendazole 2-aminosulfone及mebendazole等六種benzimidazoles之分析方法。檢體中之benzimidazoles在鹼性情況下以乙腈萃取，經Sep-Pak C18萃取匣淨化後，再以HPLC分析。結果發現利用Cosmosil 5C18 MS-II（5μm, 4.6mm i.d. × 250mm）層析管柱，以乙腈與0.02M磷酸二氫鈉溶液（pH3.3）為梯度移動相溶液，於光二極體列陣檢出器及螢光檢出器偵測下，benzimidazoles之分離效果及重複性皆佳。經以0.05～2.50μg／mL之六種benzimidazoles混合標準溶液所製作標準曲線之相關係數皆大於0.999。檢體中添加相當於0.02、0.10及1.00ppm六種benzimidazoles之平均回收率，除了低濃度之5-hydroxythiabendazole及mebendazole介於71.4～87.1%外，其餘回收率介於81.6～104.3%之間，且變異係數皆小於5.9%。本方法之最低檢出限量，紫外光檢測介於0.005～0.030 ppm，螢光檢測介於0.004～0.020 ppm，同日內及異日間之變異係數分別小於1.92%及7.22%，顯示本方法之精密度可接受。進而利用本方法分析市售豬肉、豬肝、牛肉、羊肉、牛奶及羊奶各5件，結果均未檢出benzimidazoles。