超高效液相色谱法测定牧草中马来酸麦角新碱含量

Determination of ergometrine maleate in herbage by ultra performance liquid chromatography

目的建立超高效液相色谱法(ultra performance liquid chromatography,UPLC)测定牧草中马来酸麦角新碱含量的分析方法。方法样品经纯甲醇提取后,过混合阳离子(MCX)固相萃取柱净化。采用5 mmol/L乙酸铵溶液(含0.1%甲酸)(A)和乙腈(B)作为流动相进行梯度洗脱,超高效液相色谱采用二极管阵列紫外检测器进行定量。结果本方法的检测限为0.5 mg/kg,定量限为2 mg/kg,方法在0.05~5μg/mL范围内线性良好(r=1),在2、10和20 mg/kg添加水平的回收率为83.80%~94.49%,相对标准偏差≤3.5%(n=5),实际样品测定结果为6.3~194.8 mg/kg。结论本方法具有良好的正确度和精密度,能有效测定牧草中马来酸麦角新碱含量,为将来马来酸麦角新碱应用到畜牧业打下了基础。

Objective To establish an analytical method for determining ergometrine maleate content in herbage by ultra performance liquid chromatography(UPLC).Methods The samples were extracted with pure methanol,and purified using a mixed cation(MCX)solid-phase extraction column.Gradient elution was performed using 5 mmol/L ammonium acetate solution containing 0.1%formic acid(A)and acetonitrile(B)as the mobile phases,and quantitation was performed on UPLC using a diode array UV detector.Results The detection limit and quantitation limit of the method were 0.5 mg/kg and 2 mg/kg,respectively.The linearity was good within the range of 0.05‒5μg/mL(r=1).The recoveries were 83.80%‒94.49%at the 2,10,and 20 mg/kg levels.The relative standard deviation was less than 3.5%(n=5).The actual sample determination results were 6.3‒194.8 mg/kg.Conclusion This method has good accuracy and precision,and can effectively determine the ergometrine maleate content in forage,which lays a foundation for the application of ergometrine maleate to animal husbandry in the future.