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Journal of Pharmaceutical Research

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Simultaneous Estimation of Protein and Nucleic Acid Using Derivative Spectrophotometric Method
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Journal of Pharmaceutical Research

DOI: 10.18579/jopcr/v20i3.ms21041

Year: 2021, Volume: 20, Issue: 3, Pages: 1-4

Original Article

Simultaneous Estimation of Protein and Nucleic Acid Using Derivative Spectrophotometric Method

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]

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Abstract

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

References

  1. Soh KT, Wallace PK. RNA Flow Cytometry Using the Branched DNA Technique. In: Hawley T, Hawley R., eds. Flow Cytometry Protocols. Methods in Molecular Biology. (pp. 49-77) Springer New York. 2018.
  2. Soh KT, Wallace PK. RNA Flow Cytometry Using the Branched DNA Technique. Methods in Molecular Biology. 2018;1678:49–77.
  4. Mavropoulos A, Allo B, He M, Park E, Majonis D, Ornatsky O. Simultaneous Detection of Protein and mRNA in Jurkat and KG-1a Cells by Mass Cytometry. Cytometry Part A. 2017;91(12):1200–1208. Available from: https://dx.doi.org/10.1002/cyto.a.23281
  5. Radpour R, Sikora M, Grussenmeyer T, Kohler C, Barekati Z, Holzgreve W, et al. Simultaneous Isolation of DNA, RNA, and Proteins for Genetic, Epigenetic, Transcriptomic, and Proteomic Analysis. Journal of Proteome Research. 2009;8(11):5264–5274. Available from: https://dx.doi.org/10.1021/pr900591w
  6. Dean C, Leech RM. Genome Expression during Normal Leaf Development. Plant Physiology. 1982;69(4):904–910. Available from: https://dx.doi.org/10.1104/pp.69.4.904
  11. Nevado JJB, Gallego JML, Peñalvo GC. Spectral ratio derivative spectrophotometric determination of sulphaquinoxaline and pyrimethamine in veterinary formulations. Journal of Pharmaceutical and Biomedical Analysis. 1993;11(7):601–607. Available from: https://dx.doi.org/10.1016/0731-7085(93)80011-o
  12. Pagès AM, Safdar S, Ven K, Lammertyn J, Spasic D. DNA-only bioassay for simultaneous detection of proteins and nucleic acids. Analytical and Bioanalytical Chemistry. 2021;413(20):4925–4937. Available from: https://dx.doi.org/10.1007/s00216-021-03458-6
  13. Mao X, Gurung A, Xu H, Baloda M, He Y, Liu G. Simultaneous Detection of Nucleic Acid and Protein Using Gold Nanoparticles and Lateral Flow Device. Analytical Sciences. 2014;30(6):637–642. Available from: https://dx.doi.org/10.2116/analsci.30.637

Copyright

© 2021 Published by Krupanidhi Educational Trust. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)

  • 24 November 2021

Year: 2021, Volume: 20, Issue: 3


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