@article {230, title = {Method Development and Validation for the Determination of Caffeine: An Alkaloid from Coffea arabica by High-performance Liquid Chromatography Method}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {February 2018}, pages = {88-91}, type = {Original Article}, chapter = {88}, abstract = {

Objective: The present study was investigated to develop and validate a reversed phase high performance liquid chromatography method for the determination of caffeine from bean material of Coffee arabica. Materials and Methods: The separation was achieved on a reversedphase C18 column using a mobile phase composed of water: methanol (50:50) at a flow rate of 1.0 mlmin-1. The detection was carried out on a UV detector at 272 nm. The developed method was validated according to the requirements for International Conference on Harmonisation (ICH) guidelines, which includes specificity, linearity, precision, accuracy, limit of detection and limit of quantitation. Results: The developed method validates good linearity with excellent correlation coefficient (R2 \> 0.999). In repeatability and intermediate precision, the percentage relative standard deviation (\% RSD) of peak area was less than 1\% shows high precision of the method. The recovery rate for caffeine was within 98.78\% - 101.28\% indicates high accuracy of the method. The low limit of detection and limit of quantitation of caffeine enable the detection and quantitation of caffeine from C. arabica at low concentrations. Conclusion: The developed HPLC method is a simple, rapid, precisely, accurately and widely accepted and it is recommended for efficient assays in routine work.

}, keywords = {Caffeine, Coffea arabica, Linearity, Precision, Recovery}, doi = {10.4103/pr.pr_79_17}, author = {P Naveen and HB Lingaraju and M Deepak and B Medhini and K Shyam Prasad} } @article {367, title = {Rapid Development and Validation of Improved Reversed-Phase High-performance Liquid Chromatography Method for the Quantification of Mangiferin, a Polyphenol Xanthone Glycoside in Mangifera indica}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {April 2017}, pages = {215-219}, type = {Short Communication}, chapter = {215-}, abstract = {

Mangiferin, a polyphenolic xanthone glycoside from Mangifera indica, is used as traditional medicine for the treatment of numerous diseases. The present study was aimed to develop and validate a reversed-phase high-performance liquid chromatography (RP-HPLC) method for the quantification of mangiferin from the bark extract of M. indica. RP-HPLC analysis was performed by isocratic elution with a low-pressure gradient using 0.1\% formic acid: acetonitrile (87:13) as a mobile phase with a flow rate of 1.5 ml/min. The separation was done at 26{\textdegree}C using a Kinetex XB-C18 column as stationary phase and the detection wavelength at 256 nm. The proposed method was validated for linearity, precision, accuracy, limit of detection, limit of quantification, and robustness by the International Conference on Harmonisation guidelines. In linearity, the excellent correlation coefficient more than 0.999 indicated good fitting of the curve and also good linearity. The intra- and inter-day precision showed \<1\% of relative standard deviation of peak area indicated high reliability and reproducibility of the method. The recovery values at three different levels (50\%, 100\%, and 150\%) of spiked samples were found to be 100.47, 100.89, and 100.99, respectively, and low standard deviation value \<1\% shows high accuracy of the method. In robustness, the results remain unaffected by small variation in the analytical parameters, which shows the robustness of the method. Liquid chromatography{\textendash}mass spectrometry analysis confirmed the presence of mangiferin with M/Z value of 421. The assay developed by HPLC method is a simple, rapid, and reliable for the determination of mangiferin from M. indica.

}, keywords = {Liquid chromatography{\textendash}mass spectrometry, Mangifera indica, Mangiferin, Reversed-phase high-performance liquid chromatography, Validation}, doi = {10.4103/0974-8490.204652}, author = {P Naveen and HB Lingaraju and K Shyam Prasad} }