Abstract

VALIDATED HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY METHOD FOR THE DETERMINATION OF NICOTINE IN VARIOUS TOBACCO EXTRACTS (NICOTIANA TABACUM L.)

G. Indira Priyadarshini*, Mettapalli Kavya, Nagaraju Pappula and V. Mounika

ABSTRACT

The main approach is to develop a simple, sensitive, rapid quantification of the nicotine in Nicotiana tabacum leaves by HPTLC method and its validation. Best results were obtained with Chloroform: Methanol: Liquor ammonia (6:3.5:0.5v/v/v) as the mobile phase and TLC Silica gel 60 F254 as the stationary phase to separate nicotine. The sample volume sprayed was 40μL with dosage speed of 20μL/sec, and detection wavelength was carried out at 254nm. The Rf value of Nicotine was found to be 0.824. The created method was validated for system suitability, linearity, accuracy, precision, Limit of Detection (LOD), Limit of Quantification (LOQ), and Robustness in accordance with ICH guidelines. The method was linear in the range of 4000- 8000ng/mL with a correlation coefficient of 0.9992. The % recovery was found to be 99.998% and the % RSD and precision was NMT 2. As a result, the developed methodologycan be used to conduct routine analysis.

Keywords: Nicotiana tabacum, Nicotine, HPTLC, validation, LOD, LOQ, ICH.