Durix A, Jaussaud P, Garcia P, Bonnaire Y, Bony S
INRA-ENVL Research Laboratory of Comparative Metabolism and Toxicology of Xenobiotics, National School of Veterinary Medicine of Lyon, Marcy l'Etoile, France.
J Chromatogr B Biomed Sci Appl. 1999 Jun 11;729(1-2):255-63. doi: 10.1016/s0378-4347(99)00166-8.
A high-performance liquid chromatographic method for the determination of the mycotoxin ergovaline in goat's milk is described here. Ergotamine was used as an internal standard. For a sample size of 5.0 ml, the cleanup method included precipitation of milk protein with acetone. Then, ergovaline was extracted twice with chloroform and purified by elution on an Ergosil column. HPLC separation of the extract was accomplished on a C18 column: an isocratic elution, using acetonitrile-ammonium carbonate, was performed, and the analyte was detected by fluorimetry. The method was found to be linear between 0.7 and 8 ng ml(-1), a mean recovery rate of 99.8% was obtained, and the described assay appeared both repeatable and reproducible. The limit of detection and the limit of quantitation of ergovaline in milk were 0.2 ng ml(-1) and 0.7 ng ml(-1), respectively. In order to apply the proposed method, four lactating goats were administered the toxin intravenously at a dose of 32 mg kg(-1) body weight. The concentrations of the drug in plasma and milk were then determined at standardized intervals. Ergovaline (unequivocally identified by LC-MS-MS) could not be detected in the milk beyond eight hours post-dosing. Therefore, in goats, milk does not appear to be a major excretion route for the unmetabolized toxin.
本文描述了一种用于测定山羊奶中霉菌毒素麦角缬氨酸的高效液相色谱法。以麦角胺作为内标。对于5.0 ml的样品量，净化方法包括用丙酮沉淀乳蛋白。然后，用氯仿对麦角缬氨酸进行两次萃取，并通过在麦角生物碱硅胶柱上洗脱进行纯化。提取物的高效液相色谱分离在C18柱上完成：采用乙腈 - 碳酸铵进行等度洗脱，并用荧光法检测分析物。该方法在0.7至8 ng ml⁻¹之间呈线性，平均回收率为99.8%，且所描述的测定方法具有可重复性和再现性。牛奶中麦角缬氨酸的检测限和定量限分别为0.2 ng ml⁻¹和0.7 ng ml⁻¹。为了应用所提出的方法，对四只泌乳山羊静脉注射剂量为32 mg kg⁻¹体重的毒素。然后在标准化间隔时间测定血浆和牛奶中药物的浓度。给药后八小时以上，牛奶中未检测到麦角缬氨酸（通过液相色谱 - 串联质谱法明确鉴定）。因此，在山羊中，牛奶似乎不是未代谢毒素的主要排泄途径。
