Planta Med 2011; 77 - P_69
DOI: 10.1055/s-0031-1273598

Simultaneous Determination of Tannins and Saponins from Fruits of Various Species of Terminalia using HPLC-UV, UPLC-UV-MS and HPTLC Methods

CS Rumalla 1, B Avula 1, YH Wang 1, TJ Smillie 1, IA Khan 1, 2, 3
  • 1National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, The University of Mississippi, MS 38677, USA
  • 2Department of Pharmacognosy, School of Pharmacy, The University of Mississippi, MS 38677, USA
  • 3Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia

Terminalia species are the rich source of tannins and many preparations of these species are used in traditional medicine as a cardiac tonic and diuretic. Different Terminalia species also have different ethno-botanical importance [1]. Analytical methods including HPLC, UPLC and HPTLC are presented for the simultaneous determination of hydrolysable tannins and five saponins from the fruits of different species of Terminalia (T. chebula, T. arjuna, T. bellirica, T. pallida). A separation by LC was achieved by using a reversed phase column, PDA detection, a water/acetonitrile mobile phase, both containing 0.05% formic acid using a gradient system and a temperature of 35°C. The four hydrolysable tannins (gallic acid (1), chebulagic acid (2), ellagic acid (3), chebulinic acid (4)) and five saponins (arjunglucoside-I (5), chebuloside II (6), bellericoside (7), arjunetin (8) and arjunglucoside-II (9)) could be separated within 45 minutes using a HPLC method and within 9.0 minutes using a UPLC method. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). The wavelength used for detection with the diode array detector was 254nm for tannins and 205nm for saponins. The developed methods are simple, economic, fast and especially suitable for quality control analysis of hydrolysable tannins and saponins in plant samples. The compounds from plant samples were identified and confirmed using UPLC-MS. A HPTLC method was also developed for the chemical fingerprint analysis of Terminalia species.

Figure 1: UPLC Chromatograms of a mixture of standards and Fruits of Various Terminalia species

Figure 2: HPLC Chromatograms of a mixture of standards and Fruits of Various Terminalia species at 254nm

Acknowledgements: This research is supported in part by „Science Based Authentication of Dietary Supplements“ and „Botanical Dietary Supplement Research“ funded by the Food and Drug Administration grant numbers 5U01FD002071–10 and 1U01FD003871–02, and the United States Department of Agriculture, Agricultural Research Service, Specific Cooperative Agreement No. 58–6408–2-0009.

References: [1] Kirtikar KR, et al., (2001) Indian Medicinal Plants Oriental Enterprises, 2nd edn, Uttranchal, India, 5: 1415–1439.