液相色谱-串联四极杆质谱法检测鸡配合饲料中25羟基维生素D3含量

Determination of 25-hydroxyvitamin D3 in compound feed by liquid chromatography-tandem mass spectrometry

中文关键词:  液相色谱-串联质谱  鸡配合饲料  25羟基维生素D3

英文关键词:liquid chromatography-tandem mass spectrometry  compound feed  25-hydroxyvitamin D3

中文摘要:

      目的 建立配合饲料中25羟基维生素D3的液相色谱-串联四极杆质谱(liquid chromatography-tandem mass spectrometry, LC-MS/MS)检测分析方法。方法 取适量鸡配合饲料样品于离心管中, 加水后置于振荡器上振荡破除环糊精包囊, 再加入叔丁基甲醚振荡对目标物进行提取, 在9000 r/min条件下离心5 min, 氮气吹干后用1.00 mL流动相溶解, 过滤膜后上机测定。实验使用反相色谱柱进行分离, 流动相为甲醇(0.1%甲酸)和0.1%甲酸, 采用梯度程序进行洗脱, 三重四极杆质谱进行定性定量分析, 采用内标法进行计算。结果 实验结果表明, 方法的定量限为5 μg/kg, 含量在2~500 μg/L范围内线性良好(r2≥0.9999), 加标回收率在81.18%~97.94%之间, 相对标准偏差(relative standard deviation, RSD)≤9.95%, 实际样品的含量为7.00~46.05 μg/kg。结论 本方法具有良好的准确度与精密度, 具有较好的实用性和专属性。

英文摘要:

      Objective To establish a method for the determination of 25-hydroxyvitamin D3 in compound feed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods An appropriate amount of chicken compound feed samples were placed in the centrifuge tube, water was added and placed on the oscillator to vibrate and break the cyclodextrin cyst, then tert-butyl methyl ether was added to extract the target by oscillation, and centrifuged at 9000 r/min for 5 min. After nitrogen was blow-dried, it was dissolved in 1.00 mL mobile phase, and the solution was filtered by film and then injected. Reverse phase chromatography column was used for separation. The mobile phase was methanol (0.1% formic acid) and 0.1% formic acid. Gradient program was used for elution, triple quadrupole mass spectrometry was used for qualitative and quantitative analysis, and internal standard method was used for calculation. Results The results showed that the limit of quantitation was 5 μg/kg. 25-hydroxyvitamin D3 had good linear relationships in the range of 2?500 μg/L, and the correlation coefficients were all above 0.9999, the recoveries were 81.18%?97.94%, and the relative standard deviations(RSD) were lower than 9.95%. The concentrate of actual sample were 7.00?46.05 μg/kg. Conclusion This method showed the good accuracy and precision, and had good practicability.