毛細管電泳場放大進樣──反向遷移膠束在線富集分析鯰魚和鯉魚中的孔雀石綠、結晶紫及其代謝產物   全文下載

Analysis of Malachite Green, Gentian Violet and Their Leuco Metabolites in Catfish and Carp by Micellar Electrokinetic Capillary Chromatography

本文以孔雀石綠、無色孔雀石綠、結晶紫和無色結晶紫為研究對象，開展了毛細管電泳反向膠束遷移在線富集方法的研究。結果表明，該方法具有簡單、重現、靈敏等優點。通過對分離和富集條件的優化，研究建立了毛細管電泳反向遷移膠束在線富集技術測定上述4種化合物的方法。通過對SDS 、有機改性劑濃度、水塞注射時間、進樣電壓對分離效率和富集效率的影響進行了調查。確定了緩衝液組成為50 mmol/L磷酸-140 mmol/L SDS-15% (v/v) 乙?-15% (v/v)異丙醇(pH = 2.0)的最佳分離條件，和- 8 kV電壓下壓力進樣，進樣時間為15秒，進樣前註入水柱2 s，樣品溶液基質為5 mM SDS的最佳富集條件。優化條件下，4種分析物富集率分別為121、 146、150 和87。利用該方法測定了對加標樣的?魚和鯉魚中的孔雀石綠、無色孔雀石綠、結晶紫和無色結晶紫。

A simple, reproducible and sensitive micellar electrokinetic chromatography (MEKC) method was developed for the separation and determination of malachite green (MG), leucomalachite green (LMG), gentian violet (GV) and leucogentian violet (LGV) from catfish and carp. Field-enhanced sample injection with reverse migrating micelles (FESI-RMM) was used for on-line concentration of the analytes. The buffer contained 50 mM H3PO4-NaOH, 140 mM sodium dodecyl sulfate (SDS), 15% acetonitrile and 15% 2-propanol, at the pH of 2.0. The sample solution was diluted with water containing 5 mM SDS and injected for 15 s with -8 kV after injection of 2 s water plug. The effects of concentration of SDS and organic modifier, the sample matrix, the injection time of water plug, the injection voltage and injection time of sample on the separation and stacking efficiency were investigated. Under the optimum conditions, the analytes were well separated and by optimizing the stacking conditions, about 121, 146, 150 and 87 fold improvement in the detection sensitivities were obtained for GV, MG, LMG and LGV, respectively. The instrument detection limits (S/N = 3) of GV, MG, LMG and LGV were 40, 70, 50 and 50 ng/mL, respectively. The recoveries of GV, MG, LMG and LGV in the extracts of catfish were 87.8 - 96.7%, 87.9 - 100.2%, 96.7 - 101.4% and 89.8 - 94.2%, respectively. The GV, MG, LMG and LGV in spiked (1 µg/mL) catfish and carp extracts were successfully determined with satisfactory repeatability and recovery.