@article{Shantaram_2022, title={Assessment of Different Urinary Protein Precipitation Methods for Protein Profiling by Mass Spectrometry}, volume={15}, url={https://www.asiapharmaceutics.info/index.php/ajp/article/view/4242}, DOI={10.22377/ajp.v15i04.4242}, abstractNote={<p>Aim: Urine has become one among the foremost engaging bio-fluids in clinical proteomics because is to often<br />obtained non-invasively in massive quantities and is stable compared to other bio-fluids. The study is mainly<br />focused to determine the best method for urinary protein extraction. Materials and Methods: Urinary proteins were<br />precipitated using; chloroform/methanol, acetone, and TCA/acetone solvents followed by liquid chromatography–<br />mass spectrometry (LC-MS) analysis. Results: In the present study, we have tested three different methods for urine<br />protein precipitation and qualitatively evaluated by LC-MS-analysis. The protein samples were prepared using<br />acetone, TCA/acetone, and M/C precipitation showed 51, 86, and 114 proteins, respectively. Of the percentage of<br />identified proteins by mass spectrometric analysis revealed that about 17.6% of proteins were found in all the three<br />methods. The highest percentage of shared proteins was observed between acetone and M/C precipitation (69%),<br />followed by 63.8% and 52.7% in TCA/acetone and M/C and TCA/acetone precipitation methods, respectively. By<br />contrast, M/C, TCA/acetone, and acetone precipitations showed 26%, 11.7%, and 12.4% of the unique proteins<br />excluding shared proteins, respectively. The comparison between two different methods, acetone, and M/C, showed<br />the highest percentage of shared protein as 69%, TCA/acetone and M/C precipitation showed 63.8%, and acetone<br />and TCA/acetone precipitation showed 52.7%. M/C, TCA/acetone, and acetone precipitations showed 26%, 11.7%,<br />and 12.4% of the unique proteins excluding shared proteins, respectively. Conclusion: This study contributes<br />to establish a standard procedure in urine proteomics. Using urine biomarkers, it can be widely used in urine<br />proteomics not only for diagnosis but also in basic biomedical research, such as physiology and pharmacology.</p>}, number={04}, journal={Asian Journal of Pharmaceutics (AJP)}, author={Shantaram, Manjula}, year={2022}, month={Jan.} }