Simultaneous Determination of Asaranin and Sesamin in Piper chaba Fruit by using HPTLC-MS Method: Effect of Different Extraction Methods on the Yield of Marker Compounds

Abstract

A simple, rapid and accurate HPTLC-MS method was developed for the simultaneous quantification of asaranin (AS) and sesamin (SM), the two epimeric furofuran lignans in Piper chaba (Piperaceae) fruit extracts obtained by cold percolation (PER), Soxhlet extraction (SOX), ultrasound assisted extraction (USAE), accelerated solvent extractor (ASE) and microwave assisted extraction (MAE). Separation of AS and SM was achieved on 20×10 cm pre-coated silica gel 60 F254S HPTLC plates with ethyl acetate: hexane (30:70) as mobile phase. Detection and quantification were performed densitometrically at λmax 295 nm. The peak identity was confirmed by electron spray ionization (ESI) mass spectra, which showed the [M + Na]+ ions for both AS and SM at m/z 377. The method was validated according to the ICH guidelines in terms of limit of detection (LOD) and limit of quantification (LOQ), selectivity, linearity, precision, accuracy, and robustness. The results indicate that AS (2.108%) and SM (0.103%) found to present maximum in ASE extract and minimum in PER extract (AS 0.848%; SM 0.048%). By considering the validation results, the method was found to be reproducible and convenient for quantitative analysis of AS and SM in different P. chaba fruit extracts.