Planta Med 2016; 82 - PA14
DOI: 10.1055/s-0036-1578629

Simultaneous Determination Of Chlorogenic Acid, Rutin, Hyperoside And Isoquercitrin From Leaf Extract Of Eleutherococcus Senticosus By UPLC-ESI-MS/MS In Rat Plasma And Its Application To A Pharmacokinetic Study

YH Meng 1, 2, CM Zhai 1, 2, LH Shi 1, XW Wang 1, J Li 1, IA Khan 2, 3, HX Kuang 1
  • 1Ministry of Education, Heilongjiang Key Laboratory of TCM Pharmacodynamic Material Base, 150040, Harbin, Heilongjiang Province, China
  • 2National Centre for Natural Products Research, University of Mississippi, Oxford, 38677, Mississippi, USA
  • 3Division of Pharmacognosy, Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, Oxford, 38677, Mississippi, USA

The rhizome of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim (ES), a well-known traditional Chinese medicine being used for thousands of years in China, has many pharmacological effects such as immune activity, anti-ischemia and anti-hypoxia, arrhythmia and improving sleep. Some research reported that the leaf of E. senticosus, especially the tender ones, contain the similar composition such as flavonoid, phenolic acids and exhibit the similar pharmacological effects with rhizome portions[1]. Chlorogenic acid, rutin, hyperoside and isoquercitrin are the principal compounds in the leaf of E. Senticosusand which have not been determined simultaneously using UPLC-MS/MS in previous studies. This study was aimed to develop and validate a fast, simple, sensitive, specific, reliable method to apply to pharmacokinetic study of the four active components after a single oral administration of E. senticosus leaf extract. The analytes and Bergenin (I.S.) were separated on the Waters ACQUITY UPLC and the detection was performed on a triple quadruple tandem mass spectrometer by multiple reactions monitoring (MRM) mode via electrospray ionization (ESI) source operating in the negative ionization mode (Figure 1) [2]. Chlorogenic acid, rutin, hyperoside and isoquercitrin and Bergenin (I.S.) were monitored on 353.2→191.1, 609.5→300.1, 463.1→300.3, 462.7→300.0 and 326.9→192.1, respectively. The linear range (ng/mL) of standard curve were 1.25 – 1200.0, 0.50 – 400.0, 1.50 – 1000.0, and 0.25 – 400.0 ng/. The current UPLC-MS/MS assay was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability and was suitable for pharmacokinetic studies of the four analytes after oral administration of the leaf extract.

Fig. 1: Representative MRM chromatograms of 1 for Bergenin, 2 for Chlorogenic acid, 3 for Rutin, 4 for Hyperoside and 5 for Isoquercitrin in (A) blank plasma, (B) blank plasma spiked with the four analytes and I.S. at LLOQ, (C) blank plasma spiked with the four analytes and I.S. (D) plasma sample from a rat 10 min after oral administration of leaf of E. senticosus.

Acknowledgements: This work was supported by the Program of Natural Science Foundation for Youth of Heilongjiang Province (No. QC2012C064)

References: [1] J.L. Jin, et al. Planta Med. 70(2004)564. [2] B. Ma et al. Chromatogr. B 917 – 918(2013)84 – 92.