Analytical validation of caffeine quantification method in decaffeinated coffee by HPLC-UV/ Validação analítica do método de quantificação de cafeína em café descafeinado por HPLC-UV
DOI:
https://doi.org/10.34119/bjhr2n5-061Keywords:
Caffeine, Decaffeinated coffee, HPLC-UV, ValidationAbstract
This study aimed to determine the caffeine content and to verify that the contents of decaffeinated coffee are within the standards determined by regulatory agencies. This paper describes a rapid (7.0 min) and sensitive (LLOQ 0.8 µg/mL) analytical method for the quantitation of Caffeine (CFF) in decaffeinated coffee. The method is based on High-performance Liquid chromatography-Ultraviolet (HPLC-UV) with a wavelength of 285nm. Sample preparation involved liquid-liquid extraction with chloroform. The chromatographic separation was achieved on a ACE C18 (150 x 4,6 mm) reversed-phase column and a mobile phase containing acetonitrile/water (20:80 v/v), in isocratic conditions. The assay was linear in the concentration range of 10 – 100 µg/mL. The mean recovery for CFF was 99.89%. Intra- and inter-day precision (R.S.D.) were < 1.5 % and <1.82 %, respectively and the accuracy (R.E.) was in the range ± 2.14 %.The results, verified by the validation, showed an adequate technique to be applied in the samples evaluated, in addition to a simple, fast execution methodology, with lower costs.
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