Gas Chromatography – Mass Spectrometry Method for Separation, Identification and Quantification of Β- Sitosterol, Lupeol and Friedelin from Nyctanthes Arbor-tristis and Its Marketed Formulation

In the present work a simple, precise, accurate and reproducible Gas Chromatography-Mass Spectrometry method is developed and validated for separation, identification and quantification of three pharmacologically active compounds, β-sitosterol, Lupeol and Friedelin simultaneously in the methanolic leaf extract of Nyctanthes arbor-tristis and from its marketed formulation. These compounds were extracted from the Nyctanthes arbor-tristis leaves and from its formulation using Soxhlet extraction followed by chromatographic separation using gas chromatography instrument coupled with mass spectrometer for online identification. The method was developed using Restek RTX-5MS column which gave good resolution for the components with retention time of 14.540 min, 16.257 min and 19.357 min for β-sitosterol, Lupeol and Friedelin respectively. The total run time for the programme was 21.67 min. The quantity of β-sitosterol, Lupeol and Friedelin in leaves of Nyctanthes arbor-tristis was found to be 0.074377 %, 0.03295 % and 0.010902 % respectively, while in the formulation it was found to be 0.0085548 %, 0.003384 % and 0.0028246 % respectively. Friedelin was linear between 1 – 30 μg/mL, Lupeol was linear between 1 – 50 μg/mL range & β-sitosterol was found to be between 10 120 μg/mL. The method showed recovery within the acceptable range for all the three standards. Recovery for β-sitosterol, Lupeol and Friedelin was found to be 93.12 %, 98.78 % and 105.34 % respectively in the plant, whereas in the case of formulation it was found to be 98.24%, 102.93 % and 107.56 % respectively. The RSD for precision and reproducibility calculated were below 2%. This method can be used as a quality control tool for standardization of plant material and formulation containing its extract.