DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR QUANTITATIVE ESTIMATION OF alpha-MANGOSTIN IN THE RIND EXTRACT AND FRACTIONS OF GARCINIAMANGOSTANA L. AND THEIR CYTOTOXIC ACTIVITY ON T47D BREAST CANCER CELL LINE

  • Regina Andayani Department of Pharmacy, Faculty of Pharmacy, Andalas University, Kampus Limau Manis, Padang, West Sumatra, Indonesia, 25163
  • Fatma Sri Wahyuni Department of Pharmacy, Faculty of Pharmacy, Andalas University, Kampus Limau Manis, Padang, West Sumatra, Indonesia, 25163
  • Yan Wirasti Department of Biomedic, Faculty Medicine, Andalas University, Kampus Limau Manis, Padang, West Sumatra, Indonesia, 25163
  • Dachriyanus . Faculty of Pharmacy, Andalas University, Kampus Limau Manis, Padang, West Sumatra, Indonesia, 25163

Abstract

Objective: To develop a cheap, accurate, precise, linear and rapid Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method and validate as per ICH guidelines for the quantitative estimation of α-mangostin in the rind extract and fraction of mangosteen (Garcinia mangostana L.) as well as to determine their cytotoxic activity against T47D breast cancer cell line.

Methods: The optimized method uses a reverse phase column, Shimadzu ®Shimp-pack VP – ODS (4.6 x 250 mm; 5µ), a mobile phase of 0.1 % v/v H3PO4 in water: acetonitrile (15:85), flow rate of 1 ml/min and a detection wavelength of 243,2 nm using a UV detector. The cytotoxic activity against breast cancer cell line T47D was determined as percentage of cell viability by using MTT (Microculture Tetrazolium Assay) colorimetric assay and IC50(concentration that inhibits cell growth by 50%) were calculated.

Results: The developed method resulted in α-mangostin eluting at 8.87 min. α-Mangostin exhibited linearity in the range 0.5 – 30 μg mL-1, and precise (intra-day variation ≤ 0.10 %, inter-day variation ≤ 2.28 %). The average percentage mean recovery was 94.41-102.01 %, during accuracy studies. The limit of detection (LOD) and limit of quantification (LOQ) was found to be 0.2807 and 0.9357 μg mL-1 respectively. The concentration of α-mangostin in the 70% ethanol extract, n-hexane fraction, ethyl acetate fraction and n-butanol fraction were 50.73; 11.12; 98.66; 2.29 % w/w, respectively. This extract and fractions had IC50 of 1.375; 5.879; 0.463; and 51.839 μg mL-1 against T47D cell line respectively.

Conclusion: A cheap, accurate, precise, linear and rapid RP-HPLC method was developed and validated for the quantitative estimation of a-mangostin in the rind extract and fraction as per ICH guidelines and hence it can be used for the quality control of crude extract and herbal formulation. The strongest cytotoxic activity was showed by an ethyl acetate fraction of fruit rind of Garcinia mangostana L, against T47D cell line. These results were in agreement with the concentration of α-mangostin.

 

Keywords: Garcinia mangostana L, Mangosteen, RP-HPLC, Cytotoxic activity, T47D cell line, MTT assay

Downloads

Download data is not yet available.

References

1. Obolskiy D, Pischel I, Siriwatanametanon N, Heinrich M. Garcinia mangostana L: a phytochemical and pharmacological review. Phytother Res 2009;23(8):1047-65.
2. Bennett GJ, Lee HH. Xanthones from guttiferae. 
Phytochem 1989;28:967–98.
3. Suksamrarn S, Suwannapoch N, Ratananukul P, Aroonlerk N, 
Suksamrarn A. Xanthones from the green fruit hulls of Garcinia 
mangostana. J Nat Prod 2002;6:761–3.
4. Mahabusarakam W, Iriyachitra P, Taylor WC. Chemical constituents of Garcinia mangostana. J Nat Prod 1987;50:474–8.
5. Gopalakrishnan G, Shankaranarayanan D, Kameswara L, Nazimudern SK. Effect of mangostin, a xanthone from Garcinia mangostana Linn in immunopathological and inflammatory reactions. Indian J Exp Biol 1980;18:843–6.
6. Iinuma M, Tosa H, Tanaka T, Asai F, Kobayashi Y, Shimano R, Miyauchi K. Antibacterial activity of xanthones from guttiferaeous plants against methicillin-resistant Staphylococcus aureus. J Pharm Pharmacol 1995:48:861–5.
7. Suksamrarn S, Suwannapoch N, Phakhodee W, Thanuhiranlert J, Ratananukul P, Chimnoi N, et al. Antimycobacterial activity of prenylated xanthones from the fruits of garcinia mangostana. Chem Pharm Bull 2003;51:857–9.
8. Moongkarndi P, Kosem N, Kaslungka S, Luanratana O, Pongpan N, Neungton N. Antiproliferation, antioxidation and induction of apoptosis by Garcinia mangostana (mangosteen) on SKBR3 human breast cancer cell line. J Ethnopharmacol 2004;90:161–6.
9. Matsumoto K, Akao Y, Kobayashi E, Ohguchi K, Ito T, Tanaka T, et al. Induction of apoptosis by xanthones from mangosteen in human leukemia cell lines. J Nat Prod 2003;66:1124–27.
10. Yodhnu S, Sirikatitham A, Wattanapiromsakul C. Validation of LC for the determination of α-mangostin in mangosteen peel extract: a tool for quality assessment of garcinia mangostanaL. J Chromatogr Sci 2009;47:185-7.
11. Pothitirat W, Gritsanapan W. Thin layer chromatography densitometric analysis of α-mangostin content in Garcinia mangostana fruit rind extracts. J AOAC Int 2008;91:1145-8.
12. International conference on harmonization of note for guidance on validation of analytical procedures: text and methodology, Step 5 of the ICH Process; 1995.
13. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assay. J Immunol Methods 1983;65:55-63.
14. Boik J. Natural compounds in cancer therapy. 1st ed. Oregon Medical Press, LLC, Princeton, Minnesota, USA; 2001. p. 6-8.
Statistics
369 Views | 920 Downloads
How to Cite
Andayani, R., F. S. Wahyuni, Y. Wirasti, and D. . “DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR QUANTITATIVE ESTIMATION OF Alpha-MANGOSTIN IN THE RIND EXTRACT AND FRACTIONS OF GARCINIAMANGOSTANA L. AND THEIR CYTOTOXIC ACTIVITY ON T47D BREAST CANCER CELL LINE”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 7, no. 2, Dec. 2014, pp. 174-8, https://innovareacademics.in/journals/index.php/ijpps/article/view/3638.
Section
Original Article(s)