Validated UPLC/MS Method for the Determination of Salannin in Nimbatiktum (Anti Psoriatic Herbal Formulation)

Abstract

Neem oil extract (Nimbatiktum) is a yellowish solid alcoholic extract obtained from the seed oil of Azadirachta indica of family Meliaceae. It has been used for management of psoriasis in the ayurvedic system of medicine and reported to possess anti-arthritic [1] and anti-inflammatory activity [2]. In the present investigation, a rapid, sensitive fast and high throughput validated ultra-high performance liquid chromatography tandem mass spectrometry (UPLC/Q-TOF-MS) method was established for plasma analysis of salannin for this anti-psoriatic extract. The chromatographic separation was achieved on a Waters ACQUITY UPLCTM BEH C18 (100.0mm × 2.1mm; 1.7 µm) column using gradient mobile phase, consisting of acetonitrile and 1mM ammonium acetate, at a flow rate of 0.25 mL min-1. The Waters Q-TOF premier was operated in multiple reactions monitoring mode via positive ionization interface using the transitions m/z 619.37→459.30 for salannin. The recovery of the analytes from Wistar rat plasma was optimized using liquid-liquid extraction technique (LLE) in ethyl acetate. The total run time was 2.0min and the elution of salannin occurred at 1.25±0.04min. The linearity was established over the concentration range 1–1000ng mL-1 for salannin. The lower limit of quantitation for salannin was 1.0ng mL-1. The applicability of this assay was demonstrated and successfully applied for pharmacokinetic profiling in Wistar rat. Various pharmacokinetic parameters like tmax (1.5h), Cmax (716.8µg/mL), AUC (1521.86) and t1/2 (4.4h) were calculated by taking peak plasma concentration of salannin at different time intervals. Acknowledgement: Authors are thankful to CCRAS, for financial assistance.