Identification and characterization of Ethyl gallate from Ethyl acetate fraction of Phyllanthus emblica fruit, and invitro free radical scavenging activity

Phyllanthus emblica fruit has a rich source of polyphenolic compounds and is widely used due to its medicinal properties. A portion of the ethyl acetate fraction (EAF) from 70% methanol extract (70% ME) showed greater 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging activities. Moreover, EAF has more polyphenol content than 70% ME. The EAF was subjected to a dry silica gel column eluted successively with a mixture of chloroform: ethyl acetate: formic acid (2.8: 2.4: 0.4ml) as a mobile phase solvent system, and to afford various fractions. EAF and isolated fraction-1 of ethyl gallate (EG) showed a unique pattern of EG standard chromatogram in thin layer chromatography (TLC) analysis. EAF and isolated fraction-1 real time (R.T) of the peaks were unique compared with those of the standard chromatogram in high-performance liquid chromatography (HPLC) analysis. This chromatogram fraction was collected from isolated fraction-1 at the same R.T time by multiple HPLC analysis. The collected fraction from HPLC was injected into Mass spectroscopy (MS) and it showed a molecular ion at m/z 197 corresponding to C9H10O5. The substance was identified as EG based on the results mentioned above

A novel RP-HPLC method for quantification of cholinesterase activity in human blood: An application for assessing organophosphate and carbamate insecticide exposure.

Several methods have been reported to estimate Acetylcholinesterase (AChE) enzyme activity in blood samples. The Ellman assay is the most important among all but with several shortcomings, and there is a need to develop a method which is accurate, sensitive and quick for analyzing AChE. Therefore, we have developed an assay utilizing RP-HPLC with UV detection for the determination of AChE activity. This method measured the conversion of 1-naphthol acetate to 1-naphthol to estimate AChE activity in blood samples. Performance was judged on the basis of reproducibility, sensitivity, accuracy, and the ability to screen enzyme activity within 20minutes. A series of experiments were performed, varying the concentration of blood and substrate, with optimal sensitivity using 50 μM substrate and 10μL blood. The validation parameters such as linearity (R2 of ≥ 0.9842 for 1-naphthol and ≥ 0.9897 for 1-naphthol acetate), precision (94.21-96.41%), accuracy (85.2%-99.6% and 82.6%-99.3% for 1-naphthol and 1-naphthol acetate respectively), and robustness were validated according to International Conference on Harmonization (ICH) guidelines. Blood samples were collected from healthy people, farmers exposed to spraying of pesticides, and suicidal patients who ingested pesticides and were hospitalized and were analyzed by the developed method. The AChE level was approximately 21 units/mL compared to 24units/mL in controls, whereas suicidal patients showed the least AChE levels of 1 unit/mL. The employment of this method is recommended for estimating AChE level on various matrices

Synthesis of some novel orsellinates and lecanoric acid related depsides as <i>α</i>-glucosidase inhibitors

<p>Sixteen novel orsellinic esters (<b>6a-l, 7a-d</b>) along with four lecanoric acid related depsides (3<b>a-c, 4</b>) were synthesized and confirmed their structures by spectroscopic data (¹H, ¹³C & HRMS). The synthesized compounds were evaluated for their <i>in vitro α</i>-glucosidase (<i>Saccharomyces cerevisiae</i>) inhibitory potential. Among the tested compounds, <b>3c</b> (IC₅₀: 140.9 μM) and <b>6c</b> (IC₅₀: 203.9 μM) displayed potent α-glucosidase inhibitory activity and found more active than the standard drug acarbose (IC₅₀: 686.6 μM). Both the test compounds were subjected to <i>in vivo</i> antihyperglycemic activity using sucrose loaded model in Wistar rats and found compound <b>3c</b> exhibited significant reduction in glucose levels.</p