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《Progress in Veterinary Medicine》 2018-05
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Determination of Altrenogest Concentration in Porcine Plasma with UPLC-MS/MS

XIA Liang-you;LI Shi-yang;WANG Shuang;LI Yu-chen;LI Long-fei;YANG Hai-feng;BU Shi-jin;Veterinary Medicine College,Yangzhou University;  
In order to establish a UPLC-MS/MS method for the determination of the content of altrenogest in porcine plasma,extraction of plasma was conducted with altrenogest liquid liquid extraction method,using Inertsil C4(3×150 mm,5 m)to test the separation of each component in the sample column and gradient elution,multiple reaction monitoring three level four pole mass spectrometer(MRM)model for quantitative analysis,internal standard method.The mobile phase A was 5 mmol/L ammonium formate solution,B was acetonitrile,the flow rate was 0.4 ml/min,the sample size was 10μL.In this method,the linear relationship between the concentration of 1 ng/mL-100 ng/mL and the correlation coefficient was0.999 2.Altrenogest three blank plasma concentration samples(1 ng/mL,50 ng/mL,100 ng/mL)of the batch recovery rate were 93.1%±3.98%,92.7%±5.75%,88.9%±3.69%,respectively,the intra day coefficients of variation were 3.50%-14.35%,3.43%-6.20% and 2.88%-4.15%,respectively,the coefficient of variation between 5.11%-8.86% in the daytime.The detection limits(LOD)and the quantitation limit(LOQ)of this method were 0.5 ng/mL and 1 ng/mL,respectively.The effect of matrix on the determination of the content of altrenogest was enhanced,while the low concentration(1 ng/mL)was 114.65%,the concentration of medium(50 ng/mL)was about 109.23%,and the high concentration(100 ng/mL)was113.59%.When the sample was pretreated,the coefficients of variation of the low,middle and high concentration levels were less than 11%in the sample room(8 h)for 4℃.After repeated freezing and thawing for three times,the coefficients of variation of the low,middle and high concentration levels were less than 9%,which indicated that the plasma samples could maintain high stability after being frozen and thawed for 3 cycles.The method can be used for the accurate determination of the content of altrenogest in porcine plasma.
【Fund】: 江苏高校优势学科建设工程(PAPD)
【CateGory Index】: S859.7
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