Validation of Analytical Method for Determination of Adenine and Hypoxanthine Purine Bases in Melinjo Chips by HPLC-UV

Hanifah Nuryani Lioe, Dahrul Syah, Mutiara Pratiwi, Annisa Defriana

2019

Abstract

Melinjo chips, which is commonly consumed by Indonesian people, are considered as one of the causes which triggers gout disease due to its purine content. The method to analyze purine in food is limitedly known by food laboratories in Indonesia. The objective of this research was to validate the analytical method for purine bases determination in melinjo chips by HPLC-UV. Adenine and hypoxanthine were of the known purine bases and chosen to be analyzed due to their characteristics which cause more uric acid accumulation in the body rather than other purine bases, guanine and xanthine. Guanine and xanthine were insoluble in the mobile phase used in this study, so that they might not be able simultaneously analyzed with adenine and hypoxanthine. Adenine and hypoxanthine standards were used in the instrumental performance experiment, method linearity and recovery test. The results showed that the HPLC-UV instrument with RP-C18 column and UV 257 nm detection had a good linearity in the concentration range of 7.81–125.00 µg/mL. The coefficients of determination (R2) were more than 0.999 for both adenine and hypoxanthine. Adenine and hypoxanthine were detected by HPLC-UV at retention time of 5.9–6.8 and 4.8–5.5 min respectively, and both retention times had an acceptable precision, less than 2.0 %. Detection limit (LOD) and quantification limit (LOQ) of the instrument were found at 0.72 and 2.39 µg/mL for analysis of adenine, while for analysis of hypoxanthine were at 0.69 and 2.30 µg/mL, respectively. The analytical method showed a good linearity at a concentration range of 50–800 µg/g sample with R2 more than 0.990 for both adenine and hypoxanthine analysis. Method detection limit (MDL) of adenine and hypoxanthine analysis was 19.44 and 14.42 µg/g respectively. Accuracy of the method was determined by a recovery test at spiking concentrations of 100, 500, and 1000 µg/g. In the analysis of adenine, the respective recovery results were 79.33%, 89.39%, and 90.37% with respective precisions were 5.19%, 4.50%, and 3.46%. While in the analysis of hypoxanthine, the recovery results were 66.75%, 92.29%, and 100.15%, and the precisions were 2.98%, 3.15%, and 2.22%, respectively. Based on these results, the analytical method for determination of purine bases in melinjo chips has been validated and was found to be accurate at concentration more than 100 µg/g wet weight of sample.

Paper Citation

in Harvard Style

Lioe H., Syah D., Pratiwi M. and Defriana A. (2019). Validation of Analytical Method for Determination of Adenine and Hypoxanthine Purine Bases in Melinjo Chips by HPLC-UV. In Proceedings of the 2nd SEAFAST International Seminar - Volume 1: 2nd SIS, ISBN 978-989-758-466-4, pages 118-126. DOI: 10.5220/0009978300002833

in Bibtex Style

@conference{2nd sis19,
author={Hanifah Lioe and Dahrul Syah and Mutiara Pratiwi and Annisa Defriana},
title={Validation of Analytical Method for Determination of Adenine and Hypoxanthine Purine Bases in Melinjo Chips by HPLC-UV},
booktitle={Proceedings of the 2nd SEAFAST International Seminar - Volume 1: 2nd SIS,},
year={2019},
pages={118-126},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0009978300002833},
isbn={978-989-758-466-4},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 2nd SEAFAST International Seminar - Volume 1: 2nd SIS,
TI - Validation of Analytical Method for Determination of Adenine and Hypoxanthine Purine Bases in Melinjo Chips by HPLC-UV
SN - 978-989-758-466-4
AU - Lioe H.
AU - Syah D.
AU - Pratiwi M.
AU - Defriana A.
PY - 2019
SP - 118
EP - 126
DO - 10.5220/0009978300002833