Journal of Pharmaceutical Research
DOI: 10.18579/jopcr/v20i3.ms21041
Year: 2021, Volume: 20, Issue: 3, Pages: 1-4
Original Article
Atul Agrawal1, Kiranjot Kaur1, Rakesh Sharma1, Amit Goyal1, Arti Thakkar1,2,*
1ISF College of Pharmacy, Ferozepur Road, Ghal Kalan, Moga, 142 001, Punjab, India
2Amity Institute of Pharmacy, Amity University, Sector: 125, Noida, 201 313, Uttar Pradesh, India
*Corresponding Author email: [email protected] , [email protected]
Purpose: Simple and precise first derivative zero crossing spectrophotometric method has been developed for the simultaneous estimation of protein and nucleic acid without prior separation. Approach: The concentration ranges for protein and nucleic acid were taken in the range of 60-140 mg mL-1 and 20-100 mg mL-1 respectively in 0.1 M NaOH. Absorption spectra of the samples were recorded between 200 nm to 400 nm against a reagent blank. Zero-order spectra of protein and nucleic acid were stored individually within the above concentration ranges and were derivatized in first order using scaling factor 5 for both the substances. First derivative amplitudes were recorded at 261 nm and 289 nm for estimation of protein and RNA respectively for pure substances and binary mixture. Finding: Limit of detection was found to be 2.90 mg mL-1 and 0.36 mg mL-1 and limit of quantitation was 8.80 mg mL-1 and 1.02 mg mL-1 respectively for protein and nucleic acid respectively. Precision was found to be 1.38 % for protein and 1.25 % for RNA. Reproducibility was found to be 1.54 % for protein and 1.69 % for RNA. Conclusions: Thus, proposed method can be adapted for simultaneous determination of protein and nucleic acid and better alternate technique for immunoassays and electrophoretic methods.
Keywords: First derivative zero crossing spectrophotometry; Protein; Nucleic acid; Ribonucleic acid; Analytical method validation
© 2021 Published by Krupanidhi Educational Trust. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)
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