To develop an analytical method for determination of ginkgolic acids in the ginkgo biloba extract for injection and its preparation by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.Methods:The preparations were purified by HLB SPE.A chromatographic column，Waters Cortecs T3(50 mm×2.1 mm，2.7 μm)，was used with methanol-1% acetic acid(90:10）as the mobile phase.The ginkgo acids were detected by electrospray ionization mass spectrometry in negative mode with multiple reaction monitoring(MRM）mode.Results:Ginkgo Acid C13:0，C15:1 and C17:1 possessed good linear correlation in the mass concentration range from 0 to 50 ng/mL，with the correlation coefficients more than 0.999.The mean recoveries were in the range of 97.3%-115.2%，and the RSDs were 0.3%-3.4%.The limits of quantification were 0.03，0.06，0.04 mg/kg，respectively.Conclusion:The method could be applied to the analysis of ginkgolic acids in actual samples.