Owing to the instability of Epigallocatechin Gallate (EGCG), it may undergo auto-oxidation and form oxidised products or dimers. In the present study, we aimed to evaluate the therapeutic effects, including antioxidation and immunomodulatory action, of the Oxidised Epigallocatechin Gallate (O-EGCG) as compared to native EGCG and the action of these compounds on main protease (Mpro) docking against SARS-CoV-2. HCT-116 (Human Colon Cancer) cell lines were used to estimate the total antioxidant capacity and lipid peroxidation levels and pro-inflammatory markers (human IL-6, IL-1β, TNF-α). Further, molecular docking analysis was performed by AutoDock and visualised in Discovery studio. Improved antioxidant capacity of O-EGCG was observed, and there was a significant decrease in the inflammatory markers (IL-1β, IL-6, and TNF-α) when O-EGCG was applied as compared to EGCG. The O-EGCG was shown to be strongly associated with the highest docking score and active site residues of IL-1, IL-6, and TNF- α, as well as the Mpro of SARS-CoV-2, according to in silico approach. The in vitro and in silico analyses indicate an improved therapeutic action of the oxidised form of EGCG. The effective inhibitory action of O-EGCG against SARS-CoV-2 suggests further exploration of the compound against COVID-19 and its efficacy. However, in vivo studies and understanding of the mechanism of action of O-EGCG may yield a better opinion on the use of O-EGCG and future human clinical trials.
In this paper, 3-D discrete Hartley, cosine and Fourier transforms are used for the compression of magnetic resonance images and x-ray angiograms. The performance results are then compared and evaluated. The transforms are applied on image blocks of sizes 8x8xM where M represents the number of slices. The resultant transform coefficients are quantized and then encoded using a combination of run length and Huffman coding schemes to achieve maximum compression. The performances of the transforms are evaluated in terms of peak signal to noise ratio and bit rate. It is found from the experimental results, that 3-D discrete Hartley transform yields the best results for magnetic resonance brain images whereas for x-ray angiograms the 3-D discrete cosine transform is found to be superior to the other two transforms.
Objective
In the present study we reported oxidation of epigallocatechin-3-gallate (EGCG) and validation of oxidized product by a validated ultra high-performance liquid chromatography (UHPLC) method.
Methods
Two hundred milligrams of EGCG was oxidized in 5 mL of hydrogen peroxide (H2O2) and was identified by a validated UHPLC method with precision and robustness. Confirmation of parameters like C–H stretching and mass was carried out using infrared spectroscopy and mass spectroscopy, respectively. Identification of oxidized EGCG (O-EGCG) was done by UHPLC.
Results
The infrared spectroscopy chromatograms observed less intensity C-H stretching as compared to O-EGCG. The mass of EGCG and O-EGCG were 459.09 and 915.16, respectively. Structure elucidation revealed a loss of one proton in O-EGCG as compared to EGCG. Validation of the developed method was specific, with linear correlation coefficient 0.9981 and 0.9917, respectively for EGCG and O-EGCG, the accuracy rate of 95.2%–99.6% for EGCG, and 99.18%–101.5% for O-EGCG.
Conclusion
Together, the results of this study demonstrate the formation of a dimer also the UHPLC method developed for identification of both EGCG and O-EGCG is validated as per the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines.
A simple, accurate, precise, reproducible, highly sensitive, an economic spectrophotometric method has been developed for the estimation of finasteride. UV spectrophotometric method is based on the measurement of absorption at a maximum wavelength of 255 nm. The developed method was validated with respect to linearity, accuracy (recovery), Precision (inter and intraday variations), LOD and LOQ. Beer’s law was obeyed in the concentration range of 5 – 25 μg/mL with a correlation coefficient of 0.9933. Results of the analysis were validated statistically and by recovery study. Hence the developed and validated method can be used for estimation of finasteride.
Although Epigallocatechin gallate (EGCG) is the most available and beneficial catechin found in tea, its auto-oxidation property may lead to toxicity when consumed in large quantities. Thus, there is a need to quantify the EGCG, which enables to study the pharmacological characteristics of the compound. The study aimed to develop and validate a rapid and accurate analytical method for quantitative determination of EGCG. Standard EGCG was used to conduct trials for the optimization of the analytical method using Ultra-High Performance Liquid Chromatography (UHPLC). Tests for validation (specificity, linearity, accuracy, system suitability, method precision, robustness, and ruggedness) were performed. The preliminary trials yielded an analytical method with good peak shape and acceptable system suitability which was further validated. The method was shown to be specific, with a linear correlation coefficient of > 0.9996 and accurate with acceptable recovery rate (99.1% to 100.4%). Acceptable system suitability and method precision were confirmed with a relative standard deviation (less than 2%). Further, robustness and ruggedness experiments also demonstrated the suitability of the present analytical method. The method developed for determination of EGCG was validated as per the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and thus can be used in routine compliance tests in the laboratory for further studying/characterizing the properties of EGCG.
OPEN ACCESSCitation: U. V. R, R. SS, Kumar K. R, Narayan Sinha S (2020) Method development and validation for rapid identification of epigallocatechin gallate using ultra-high performance liquid chromatography. PLoS ONE 15(1): e0227569.
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