Establishment of analytical methods for nucleobase and nucleosides of leech formula granule by LC-MS/MS
An analytical method was established for simultaneous determination of 7 nucleobase and nucleoside components in leech formula granule by a combination of Shimadzu ultra-high performance liquid chromatography Nexera X2 and triple quadrupole mass spectrometry LCMS-8050.Leech formula granule was extracted by ultrasound at 50 ℃ with 10％methanol for 30 minutes,and detected with MRM mode by both positive and negative ESI ionization.Discovery®HS F5-3 column was used for separation,with a mobile phase consisted of 0.15％formic acid aqueous solution and methanol.Gradient elution was used within 10 minutes.External standard method was used for quantification.The seven compounds had good linear relationships within the corresponding concentration range,all coefficient of determination(R2)were greater than 0.996,and the quantification limit was between 0.42 μg·g-1and 3.94 μg·g-1.The 10 ng·mL-1 mixed standard solution was injected 6 times,and the peak area RSD of tested compounds was all less than 6％,indicating good precision.The recovery experiments executed for six times,the average recovery of 7 compounds was ranged from 87.09％to 105.81％,with RSD％ranging from 1.35％to 4.70％.The residual test result was negative.This method is accurate and reliable,and can provide a reference for the quality control of leech formula granule.
万方数据
维普
