Analysis of Vincristine in Leaf and Callus Extract by Validated HPTLC Technique and Its Stress Degradation Studies

Aims: An incipient, concrete, sensitive, selective, precise, and reproducible high-performance thin-layer chromatographic (HPTLC) method was established and validated for estimation of vincristine alkaloid in leaf extract as well as it’s callus.

Study Design: Mention the design of the study here.

Place and Duration of Study: Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi, India between June 2010 and July 2011.

Methodology: HPTLC was performed on aluminium foil precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of 73:25:2, v/v (methanol: toluene: diethyl amine) as a mobile phase. The densitometric analysis was carried out at 254 nm utilizing Camag TLC scanner which engendered compact spots for vincristine (Rf =0.38±0.02).

Results: The linear regression data for the calibration plots showed good linear relationship (r2=0.998±0.003) between peak area and concentration in the range of 500-3000 ng spot-1. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines. The limits of detection and quantification were 3.06 and 9.29 ng spot-1, respectively.

Conclusion: Statistical analysis showed that the method could be prosperously applied for the estimation of vincristine in leaf extract and in callus. As the method could efficaciously separate the drug from its degradation products, it can be employed as a stability indicating one. Moreover, the proposed HPTLC method was utilized to investigate the kinetics of acid degradation process.