Application of UPLC-MS/MS to simultaneously detect four bioactive compounds in the tumour-shrinking decoction (FM1523) for uterine fibroids treatment

Phytochem Anal. 2019 Jul;30(4):447-455. doi: 10.1002/pca.2827. Epub 2019 Mar 27.


Introduction: The Chinese medicine formulation, tumour-shrinking decoction (TSD, FM1523), which consists of 15 natural medicines, is used for uterine fibroids (UFs) therapy and possesses excellent clinical therapeutic effect.

Objective: To develop a sensitive and validated analytical method for the simultaneous quantification of four crucial bioactive compounds including isorhamnetin-3-O-neohesperidoside, curcumin, peimine and tetrahydropalmatine in the principal formulation of this decoction.

Methods: An ultra-performance liquid chromatography coupled tandem mass spectrometry (UPLC-MS/MS) with an electrospray ionisation (ESI) source in multiple reaction monitoring (MRM) mode was conducted to investigate these bioactive compounds in the TSD. The chromatographic separation was performed on a C18 column when the flow rate was adjusted at 0.2 mL/min with gradient elution of acetonitrile-water with 0.1% formic acid. Accelerated solvent extraction (ASE) method with higher extraction efficiency was employed for TSD sample pre-treatment.

Results: The linearity, limit of detection (LOD) and limit of quantification (LOQ) were determined for this analytical method. The mean recoveries of the compounds were determined between 100.23% and 104.02% with satisfactory relative standard deviation (RSD) in the ranges of 2.65% to 3.81%. The precision was evaluated by intra-day and inter-day tests, which revealed RSD within the ranges of 1.21% to 2.14% and 1.24% to 2.32%, respectively.

Conclusion: The bioactive compounds of TSD samples were successfully quantified via UPLC-MS/MS with MRM mode. This study could help to evaluate the pharmacokinetic study of TSD during clinical applications and present a facile strategy for quantifying bioactive compounds in traditional Chinese Medicine decoction.

Keywords: bioactive compounds; extraction; multiple reaction monitoring; tumor-shrinking decoction; ultra-pressure liquid chromatography coupled tandem mass spectrometry.

MeSH terms

  • Berberine Alkaloids / chemistry*
  • Berberine Alkaloids / isolation & purification
  • Cevanes / chemistry*
  • Cevanes / isolation & purification
  • Chromatography, High Pressure Liquid
  • Drugs, Chinese Herbal / chemistry*
  • Humans
  • Leiomyoma / drug therapy*
  • Limit of Detection
  • Phytochemicals / chemistry*
  • Phytochemicals / isolation & purification
  • Tandem Mass Spectrometry


  • Berberine Alkaloids
  • Cevanes
  • Drugs, Chinese Herbal
  • Phytochemicals
  • verticine
  • tetrahydropalmatine