Simultaneously determination of 15 bile acids in plasma of mouse by LC-MS/MS
OBJECTIVE To simultaneously determine 15 bile acids in mice plasma by UPLC-MS/MS.METHODS The analytical column was Shim-pack GIST C18(100 mm ×2.1 mm,2 μm).The mobile phase consisted of 0.01％formic acid(A)and acetonitrile with 0.01％formic acid(B)for gradient elution.The injection volume was 5 μL with a flow rate of 0.3 mL·min-1,the column temperature was maintained at 40 ℃,and the internal standard was cholic acid-d4.Monitoring with multiple reactive ions,using negative ion mode,electrospray ion source.Plasma samples were pre-treated and injected.RESULTS The 15 bile acids showed good linearity with their peak area ratios in the range of 0.24-3.2 × 104 ng·mL-1(r2≥0.9900).The recovery rate was in the range from 83％to 110％;the intra-day RSD was in the range from 0.68％to 4.02％;inter-day RSD was in the range from 1.53％to 8.06％.The recovery rate of extraction was in range of 70.24％-94.25％.The samples remained stable after repeated freeze-thaw and long-term freezing at-80 ℃.CONCLUSION The method is simple,accurate and reproducible,and can be used for the determination of 15 kinds of bile acids in mouse plasma.
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