Planta Med 2010; 76 - P35
DOI: 10.1055/s-0030-1251797

Validation of HPTLC Method for Estimation of Diosgenin in Callus and Rhizome of Dioscorea deltoidea

M Amir 1, M Mujeeb 1, A Sabih 1, S Ahmad 1, A Ahmad 1, WA Siddiqui 2
  • 1Plant tissue culture laboratory, Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi 110062, India, email aamirx1@gmail.com
  • 2Departments of Biochemistry, Faculty of Science, Jamia Hamdard, New Delhi 110062, India

Dioscorea deltoidea (Dioscoreaceae) is a rhizome found in North western Himalayas from Kashmir and Punjab to Nepal and China and some other parts of the world up to an altitude of 1000 to 3000m. Dioscorea deltoidea rhizomes contain diosgenin, which is widely used as a precursor in the synthesis of steroid hormones such as progesterone, corticosteroids, and anabolic steroids. The most important sapogenins are diosgenin and yamogenin [1]. A trouble-free, accurate, selective, precise and economical high-performance thin-layer chromatographic method for analysis of diosgenin in callus and rhizome of Dioscorea deltoidea was developed and validated. The method was developed on TLC aluminium plates precoated with silica gel 60F-254 using solvent system petroleum ether:iso-propanol (12:1, v/v), which gives compact spot of diosgenin (R f value 0.76±0.02). Densitometric analysis of diosgenin was carried out in the absorbance mode at 366nm after spraying with methanolic sulphuric acid. The linear regression analysis data for the calibration plots showed good linear relationship with r=0.991 and 0.995 for diosgenin with respect to peak height and peak area, respectively, in the concentration range 100–1,000ng per spot. The limits of detection and quantification for diosgenin were 16.58 and 50.25ng per spot, The proposed method was applied for determination of diosgenin in Dioscorea deltoidea, which showed 277.06mg/L of diosgenin. Statistical analysis proves that the method is repeatable, selective and accurate for the estimation of diosgenin in Dioscorea deltoidea. The developed method effectively resolved the diosgenin in Dioscorea deltoidea hence, it can be employed for routine analysis. Acknowledgements: The financial support from the All India Council for Technical Education (AICTE), New Delhi, India is duly acknowledged. References: [1] Hussain A, Singh P, Srivatava G.N (1979). Farm Bulletin, vol. 14, p.53–60 CIMAP, Lucknow, India.