Processing often plays an important role in determining the components and pharmacological effects of Chinese herbal medicines.In order to investigate the impact of processing on the components of Cistanche tubulosa,a high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS) method was established to simultaneously determine the content of cistanoside F,acteoside,isoacteoside,2′-acetylacteoside and echinacoside in C. tubulosa before and after processing.The medicinal powder was extracted with a 50% methanol solution.The chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C₁₈ column (2.1 mm × 150 mm,1.8 μm) using a gradient elution of 01% formic acid water (A) and acetonitrile (B) as the mobile phase.Negative ion multiple reaction monitoring (MRM) mode was selected,with calceolarioside B used as the internal standard.The sample injection volume was 1 μL,flow rate was 0.3 mL/min,column temperature was 35 ℃,and the temperature of the autosampler was set at 8 ℃ during the analysis by HPLC-QQQ-MS for the determination of these five components in C. tubulosa before and after processing in the same batch.The results showed good linearity within their respective ranges,with correlation coefficients (r) ranging from 0.992 9 to 0.998 5.The average recovery rates were between 98.15% and 100.2%,and the relative standard deviations (RSD) were less than 4.6%.The established method is simple,accurate,and can be used for quality control of C. tubulosa,as well as evaluating the impact of processing on its content and pharmacological effects.