The metabolites specifically present in eight different classes of S. asoca and two drugs were listed out. Further, the abundant metabolites which can act as representative of their groups were identified. UPLC have several advantages over the conventional techniques being a tool to give rapid and effective phytochemical fingerprints along with the quantization of Z-VAD-FMK mw marker compounds. The length of the column [250 mm] increased the column efficiency and concomitant resolution resulted separation of 4 peaks per min over a range of 4–40 min [Fig. 1]. With the help of
infused standards reproducibility of data was analyzed and retention time variability was found to be 2 s and a relative standard deviation of less than 4% was observed. Crude cold and hot water extracts of various parts of S. asoca and drugs samples were analyzed without considering any specific group of metabolites. Furthermore, no pretreatment was given to the samples to avoid discrimination and to get maximum number of metabolites. Fig. 1 shows total ion chromatograms to distinguish between bark, regenerated bark, leaves, flowers and drugs prepared from bark. A visual examination shows the differences between the samples employed in the study. Along with several unique peaks across the samples, a prominent peak at 39.9 min in the chromatograms
was observed only in regenerated bark samples [ Fig. 1A and B] which can be further exploited FK228 supplier as a marker peak of regenerating bark. Q-TOF-MS provides accurate MS/MS spectra due to internal mass calibration during acquisition and mass drift compensation. In the present study, mass accuracy less than 3 ppm was obtained when compared with internal and external standards. Q-TOF-MS was operated in positive ion mode with a ramp setting for collision energy. On-an average 8261 molecular features were observed per sample when analyzed with a threshold 5000 counts per second. Most abundant metabolites were inspected carefully and marker compounds of different parts of plants and drugs were identified [Table 1]. Some of the compounds were identified by their characteristic
mass fragments and later on comparing the m/z pattern with MS/MS spectra available with http://spectra.psc.riken.jp. One unique and un-identified metabolite of 385.9094 m/z Non-specific serine/threonine protein kinase was observed at retention time 39.98 min in the regenerated bark sample along with others described in Table 1. Prunasin was observed in both the Askokarishta samples at m/z 296.7617 with product ion m/z 276.76 due to prominent water loss from the molecule. These most abundant molecular features can be used as biomarkers of various plant parts. It also produces challenge for further research to identify these metabolites and the potential of scopes in natural product research. Furthermore, derivatives of catechin and protocatechualdehyde [data not shown] were found to be elevated during the qualitative analysis, in the re-generated bark along with feruloyl CoA.