Authors:Narendra Gajbhiye, Jayanti Makasana and Trishul Thorat
Aegle marmelos (Rutaceae) has gained traditional therapeutic importance owing to their coumarins (umbelliferone, psoralene, marmin, and imperatorin) and alkaloid (skimmianine). Presently, there is no appropriate thin-layer chromatography (TLC) based method available for simultaneous determination important coumarins and alkaloids of A. marmelos. A simple, sensitive, and accurate high-performance thin-layer chromatographic (HPTLC) method has been developed for the determination of marmin, skimmianine, umbelliferone, psoralene, and imperatorin for in the methanolic extract of root bark of A. marmelos. Analysis was performed by using TLC precoated silica gel 60 F 254 aluminium plates (20 × 10 cm) with toluene-ethyl acetate-formic acid (6:4:0.1, v/v) as mobile phase. Quantitative evaluation of the plate was performed in the absorption mode at 304 nm. The calibration curves were prepared in the concentration range of 40–500 ng spot−1 for marmin, skimmianine, and umbelliferone, for psoralene in the range of 16–200 ng spot−1 and for imperatorin in the range of 20–250 ng spot−1. The method was validated for precision, repeatability, and accuracy. The technique has been applied, for the first time, for the simultaneous estimation of marmin, skimmianine, umbelliferone, psoralene, and imperatorin. The proposed method was found to be robust, precise, and accurate; it therefore holds potential for detection, monitoring, and quantification of marmin, skimmianine, umbelliferone, psoralene, and imperatorin in A. marmelos raw root drug materials and chemical screening of germplasms.
Authors:Narendra A. Gajbhiye, Jayanti Makasana, Tushar Dhanani and Raju Saravanan
Aegle marmelos Correa (Bael tree) is a medicinal fruit tree, widely used for healing purposes in various systems of medicines. Coumarins and alkaloids present in various parts of bael tree including roots and fruit pulp are the primary active constituents implicated for its biological activities. An efficient liquid chromatography–electrospray ionization—tandem mass spectrometry (LC—ESI—MS/MS) method was developed for identification and simultaneous determination of four coumarin derivatives, namely, umbelliferone, psoralene, marmin, and imperatorin, and an alkaloid, skimmianine, in root and stem bark of A. marmelos. The chromatographic separation of analytes was performed on Altima C18 (50 × 4.6 mm, 3 μm) column using methanol and 0.1% acetic acid in water (54:46, v/v) as the mobile phase under isocratic conditions. The LC–MS/MS parameters were optimized in the positive ionization mode using electrospray ionization source. The quantification of the analytes was performed using multiple reaction monitoring (MRM) transitions, umbelliferone (m/z 163.1 → 107.1), psoralene (m/z 187.2 → 131.1), marmin (m/z 333.5 → 163.2), imperatorin (m/z 271.1 → 203.1), and skimmianine (m/z 260.1 → 227.0). The extraction method was standardized for optimum yield of coumarin derivatives and the alkaloid in different extraction solvents. Higher yield of the analytes was found in methanolic extracts in comparisons to petroleum ether, chloroform, ethyl acetate, ethanol, and water. The method was validated for linear range, intra- and inter-batch precision and accuracy. The distribution of coumarin derivatives and an alkaloid was found to vary significantly in different plant samples, and their concentration was much higher in roots as compared to stem bark.