Simultaneous determination of oligosaccharides and iridoid glycosides in rat plasma using hydrophilic interaction chromatography coupled with tandem mass spectrometry: Challenges in bioanalytical method development for pharmacokinetic study of Radix Rehmanniae extract


Xin Di, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China (dixin63@hotmail.com)


Quantitative bioanalysis of oligosaccharides by LC-MS/MS is challenging and rarely reported up to now. Simultaneous quantitative bioanalysis of oligosaccharides and iridoid glycosides by LC-MS/MS poses additional challenges due to their unique and different structural features. In the present study, a fast and sensitive analytical method based on hydrophilic interaction chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) was developed for simultaneous determination of raffinose, manninotriose, stachyose, ajugol and catalpol in rat plasma. Multiple analytical challenges were encountered during method development, including very different retention behavior of oligosaccharides and iridoid glycosides, confusing split peaks for manninotriose, low ionization and extraction efficiency of oligosaccharides, thermal instability of catalpol and reduced column performance. The strategies to overcome these challenges were presented by optimizing chromatographic separation, mass spectrometric detection and sample preparation. The best separation was achieved on an Accucore-150-Amide-HILIC column (100 mm × 2.1 mm, 2.6 μm) at 50 °C with a gradient mobile phase containing acetonitril and 2.5 mM ammonium acetate. Ammonium adduct ions produced by positive electrospray ionization were chosen as precursor ions for multiple reaction monitoring transitions. The lower limits of quantification were 0.01-0.2 mg/mL using only 50 µL of plasma sample. The method was successfully applied to pharmacokinetic characterization of oligosaccharides and iridoid glycosides in normal and type 2 diabetic rats after intragastric administration of Radix Rehmanniae extract.

Keywords: Oligosaccarides; Iridoid glycosides; Radix Rehmanniae; Hydrophilic interaction chromatography; Tandem mass spectrometry; Pharmacokinetics

Abstract Reference & Short Personal Biography of Presenting Author

Reference

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[2] Liu ZR, Lou ZY, Ding X, Li X, Qi YP, Zhu ZY, Chai YF. Global characterization of neutral saccharides in crude and processed Radix Rehmanniae by hydrophilic interaction liquid chromatography tandem electrospray ionization time-of-flight mass spectrometry. Food Chem 2013; 141:2833–2840.

[3] Xu J, Wu J, Zhu LY, Shen H, Xu JD, Jensen SR, Jia XB, Zhang QW, Li SL. Simultaneous determination of iridoid glycosides, phenethylalcohol glycosides and furfural derivatives in Rehmanniae Radix by high performance liquid chromatography coupled with triple-quadrupole mass spectrometry. Food Chem 2012; 135:2277–2286.

Dr. Xin Di obtained her BS degree in Environmental Analysis from Nanjing University of Science and Technology in 1990 and her Ph.D. degree in Pharmaceutical Analysis from Shenyang Pharmaceutical University in 1995. She has been working in Shenyang Pharmaceutical University since 1995, where she was promoted to professor in 2005. She worked as visiting research scholar in Hong Kong Baptist University from 2002 to 2003 and in Oxford University from 2007 to 2008. Dr. Di serves as the director of the Key Laboratory of Drug metabolism and Pharmacokinetics of Liaoning Province and the associate editor of Journal of Shenyang Pharmaceutical University. She is also a Distinguished Professor of Liaoning Province. Dr. Di’s research interests include drug metabolism and pharmacokinetics, development and applications of new analytical methodologies for pharmaceutical and biomedical purposes, and novel analytical strategies for fast screening of bioactive and toxic components from traditional Chinese medicines.   

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