02252nas a2200241 4500008004100000245015400041210006900195260001800264300001000282490000700292520150100299653001301800653001901813653001401832653001401846653001301860100001401873700001801887700001401905700001501919700001801934856005801952 2018 eng d00aMethod Development and Validation for the Determination of Caffeine: An Alkaloid from Coffea arabica by High-performance Liquid Chromatography Method0 aMethod Development and Validation for the Determination of Caffe cFebruary 2018 a88-910 v103 a

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.

10aCaffeine10aCoffea arabica10aLinearity10aPrecision10aRecovery1 aNaveen, P1 aLingaraju, HB1 aDeepak, M1 aMedhini, B1 aPrasad, Shyam uhttps://phcogres.com/article/2018/10/1/104103prpr7917