Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS

Abstract

Objective : To develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma. Methods ¡¤ After extraction of C4, CA and CDCA with acetonitrile from plasma, they were quantified with stand-ard curve corrected by the internal standards based on Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Results ¡¤ The limits of detection (LOD) of C4, CA and CDCA were 0.16, 0.02 and 0.04 nmol/L respectively; All three metabolites had good linear relationships (correlation coefficients R2 were over than 0.998). The relative standard deviations (RSDs) of repeatabilities were below 3.0%. The RSDs of inter-day and intra-day precision were less than 6%, and the RSDs of stabilities at 4 °C were below 10% within 7 days. The average added recoveries of C4, CA and CDCA were 97.7%, 113.3% and 105.0%, respectively. Conclusion ¡¤ This method is of high detective sensi-tivity, good precision and stability, which meets the quantitative requirements of plasma biological samples.