Planta Med 2010; 76 - P513
DOI: 10.1055/s-0030-1264811

Quality control analysis of geraniin in dried leaves of Phyllanthus muellerianus (KUNTZE) EXELL

C Agyare 1, S Rogge 1, A Baumann 1, M Lechtenberg 1, A Hensel 1
  • 1University of Muenster, Institute for Pharmaceutical Biology and Phytochemistry, Institute for Pharmaceutical Biology and Phytochemistry, Hittorfstrasse 56, 48149 Muenster, Germany

P. muellerianus is a woody climber and the leaves are used for treatment of wounds, skin eruptions, urethral discharge and dysentery [1–3]. We investigated the wound healing properties of P. muellerianus through bioguided fractionation; activity was found to be attributed to geraniin, major compound in the leaves [4]. Aims: (1) To develop simple method of extraction for P. muellerianus with maximum yield of geraniin (2) to develop and validate suitable analytical methods [5, 6] for quality control of geraniin. Methods: 1g of dried powdered leaves in 10 mL MeOH/H2O (7/3) was extracted by ultrasonic bath for 15min to yield the test solution for HPLC-analysis. Results: Different extraction procedures were compared and ultrasonic extraction was found to be the most efficient method for the extraction of geraniin. Among different solvents, MeOH/H2O (7/3) was found to bring highest yields of geraniin (>4%). Peak identification was done by spiking and co-injection with reference compound. The content of geraniin was determined to be 4.3%. Linearity and range - 6 conc. (100–1000µg/mL) showed linearity: y=384.01x –1191.1, R2 =0.9979. Accuracy and recovery: Spiking experiments were performed as follows: 6 conc., addition of geraniin from 0%-186%. The x-intercept of calibration curve (y=379.21x + 148077, R2=0.9914) gave the amount of geraniin in extract. Comparison with results obtained from standard calibration procedures gave recovery of >99%. Limit of detection: 0.5µg/mL; Limit of quantitation: 2.5µg/mL Precision: Intra-assay and intermediate precisions were 4.8 and 9.6% (% RSD) respectively.

References: 1. Burkill (1994), Useful plants of west tropical Africa, 121–122.

2. Irvine, Woody plants of Ghana (1961). 246–247.

3. Agyare et al., (2009). J. Ethnopharmacol. 125:393–403.

4. Agyare et al., (2010) Phytomedicine (submitted).

5. Text on Validation of Analytical proceedings – ICH Harmonized Tripartite Guideline, ICH; ICH-Q2A.

6. Validation of Analytical Procedures: Methodology – ICH Harmonized Tripartite Guideline, ICH; ICH-Q2B.