OPEN-ACCESS PEER-REVIEWED
1*Dr. V. Jayashree, 2Dr. Malarkodi Velraj, 3Dr. Tushar Mahendrabhai Patel, 4Parthiban S ,Mrs. 5Monisha.J, 6M. kalaivani, 7Pankaj Ramanlal Beldar
1*Associate Professor, College and university name: Vels Institute of Science, Technology and Advanced Studies
2Professor & Head, College and university: Vels Institute of Science Technology and Advanced Studies, Pallavaram, Chennai, India.
3Senior Lecturer, Department of Pharmaceutics, L. M. College of Pharmacy, Navarangpura, Ahmedabad 380009, Gujarat, India.
4Associate Professor, College and university name: Vels Medical College and Hospital, VISTAS Periyapalaya, Thiruvallur- 601102
5Assistant Professor, College and university name: Vels Institute of Science Technology and Advanced studies, Pallavaram, Chennai, India.
6Associate professor, School of pharmacy, DRIEMS University Tangi, Cuttack -754022. Odisha
7K.K.Wagh Institute of Engineering Education and Research, Nashik, Maharashtra, India.
Abstract
Therefore, results obtained from the employed analytical method revealed good linearity, precision, accuracy, recovery and sensitivity in the determination of oleic acid, palmitic acid and linoleic acid standards at a concentration range of 0.1-50 μg/mL. As shown in the figures, the calibration curves were highly linear with a coefficient of determination (R2) ranging between 0.997 to 0.999 for the three fatty acids studied. The method also showed acceptable run-to-run and batch-to-batch precision, which were in terms of relative standard deviation varying between 2.5- 5.6%. The accuracy was also 95.2 to 104.8% with the percentage recovery for the concentration range that was tested. Recoveries of the fatty acids from spiked samples did not differ significantly at low and high, spike concentrations and were at 96.8-98.4%. Furthermore, the method was found to have low limits of detection of 0.03-0.05μg/mL and a range of lower limits of quantification of 0.10-0.15μg/mL which shows that the method was sensitive. All in all, it was established that the analytical method used was highly reliable and precise in the determination of these fatty acids. It can potentially be used for regular quality control checks of the concentration of these fatty acids in infant formula, food, pharmaceuticals, cosmetics, or other related products within the validated concentration range.
Keywords: Keywords: HPLC; validation; free fatty acids; oleic acid; palmitic, acid; linoleic acid; regulatory; linearity; precision; accuracy
References
[1]. Fawzy M. A. (2017). Fatty Acid Characterization and Biodiesel Production by the Marine Microalga Asteromonas gracilis: Statistical Optimization of Medium for Biomass and Lipid Enhancement. Marine biotechnology (New York, N.Y.), 19(3), 219–231. https://doi.org/10.1007 /s 10126-017-9743-y
[2]. FDA (2024). FDA. https://www.fda.gov/ regulatory-information/search-fda-guidance-documents/m10-bioanalytical-method-validation-and-study-sample-analysis
[3]. Food and Drug Administration. (2018). Bioanalytical method validation for industry: Regulatory, application, and acceptance considerations. https://www.fda.gov/regulatory-information/search-fda-guidance-documents/bioanalytical-method-validation-guidance-industry
[4]. Friedrich, S., Antes, G., Behr, S., Binder, H., Brannath, W., Dumpert, F., Ickstadt, K., Kestler, H. A., Lederer, J., Leitgöb, H., Pauly, M., Steland, A., Wilhelm, A., & Friede, T. (2021). Is there a role for statistics in artificial intelligence? Advances in Data Analysis and Classification, 16(4), 823–846. https://doi.org/10.1007/s11634-021-00455-6
[5]. Harwood, M. D. Achour, B., Russell, M. R., Carlson, D. A., Warrack, B.M., Phinney, K. W., & Keshavarz, A. Rauh, M. (2015). Relative to these factors, the following analytical issues deserve consideration for the efficient quantification of therapeutic protein-drug byproducts by LC-MS: Journal of Pharmaceutical and Biomedical Analysis 102 (2015), pp 152-159. https://doi.org/10.1016/j.jpba.2014.09.012
[6]. International Conference on Harmonization. (1995). Validation of analytical procedures: Information and method: Written text and ICH Q2A. https://database.ich.org/sites/default/files/Q2A_Guideline.pdf
[7]. Karpe F, Dickmann JR, & Frayn K N. Fatty acids, obesity, and insulin resistance: This change can only be interpreted as a signal that it is time for a reevaluation of our strategies. Diabetes, 60(10), 2441-2449. https://doi.org/10.2337/db11-0425
[8]. Koutsari, C., Ali, A. H., Mundi, M. S., & Jensen, M. D. (2011). Storage of circulating free fatty acid in adipose tissue of postabsorptive humans: quantitative measures and implications for body fat distribution. Diabetes, 60(8), 2032–2040. https://doi.org/10.2337/db11-0154
[9]. Li J, Zhou X, Chen C, Yang X, Zheng Y, Zhang X, Xu G, Liu X. (2021) Methods used in the determination of fatty acids in biospecimens in the current world. TrAC – Trends in Analytical Chemistry, 143, 116392. https://doi.org/10.1016/j.trac.2021.116392
[10]. Makrecka-Kuka M., Krumschnabel G. & Gnaiger E., 2017 High-resolution respirometry for the parallel determination of O2 and H2O2 production in permeabilized cells, tissue homogenate, or isolated mitochondria. Biomolecules, 7(3), 13. https://doi.org/10.3390/biom7030013
[11]. Mlinar, B., Marc, J., Janež, A., & Pfeifer, M. (2007) Sempionite pathology and molecular processes that accompany insulin resistance. Clinica Chimica Acta, 375(1-2), 20-35., doi: 10.1016/j.cca.2016.09.025 https://doi.org/10.1016/j.cca.2006.07.005
[12]. Nielsen, T. S., Jessen, N., Jørgensen, J. O. L., Møller, N., & Lund, S. (2014). Dissecting adipose tissue lipolysis: Endogenous molecules and their impact on metabolism disease. J Mol Endocrinol; 32: 279–290; 52(3): R199–R222. https://doi.org/10.1530/JME-13-0277
[13]. Obanda, Derick N., Michael Downes, Aaron Lau, Heather P. Hendrickson, Kirsty Williams, Peter Katikaridis, Christian Gils, Christian Bierwirth, Sarah M. King, Richard W. Nelson, Julio Sanchez-Gurmaches, and David A. Guertin. (2021) Excess acylcarnitine accumulation within mitochondria contributes to obesity-related adverse changes in adipose tissue. Science Advances, 7(28). https://doi.org/10.1126/sciadv.abf6732
[14]. Penugonda, K., & Lindshield, B. (2013). Fatty Acid and Phytosterol Content of Commercial Saw Palmetto Supplements. Nutrients, 5(9), 3617–3633. https://doi.org/10.3390/nu5093617
[15]. Piovesana, S., Aita, S. E., Cannazza, G., Capriotti, A. L., Cavaliere, C., Cerrato, A., Guarnaccia, P., Montone, C. M., & Laganà, A. (2021). In-depth cannabis fatty acid profiling by ultra-high performance liquid chromatography coupled to high resolution mass spectrometry. Talanta, 228, 122249. https://doi.org/10.1016/j.talanta.2021.122249
[16]. Ruge, T, Lockton, JA, Renström, F, Lystig, T, Sukonina, V, Svensson, MK, Eriksson, JW, Olivecrona, G, Olivecrona, T. Increased insulin levels have an effect on plasma concentration of interleukin-6 in both non-diabetic controls and insulin-resistant obese subjects in acute hyperinsulinemic clamp. Diabetes Care, 32(7), 1376-1378. https://doi.org/10.2337/dc08-1801
[17]. Rutherfurd, S. M., & Dunn, B. M. (2011). Quantitative amino acid analysis. Current protocols in protein science, Chapter 3, 3.2.1–3.2.6. https://doi.org/10.1002/0471140864.ps0302s63
[18]. Saadatian-Elahi, M, Slimani, N., Chajès, V., Jenab, M. Goudable, J, Biessy, C., Ferrari, P., Byrnes, G., Autier, P., Peeters, P. H., Ocké, M., Bueno-de-Mesquita, H. B., & Romieu, I. Plasma phospholipid fatty acid profiles and their association with food intakes: Cross sectional survey within the framework of the European Prospective Investigation into Cancer and Nutrition. Am J Clin Nutr 2007; 86: 1476–92 . https://doi.org/10.3945/ajcn.2008.26834
[19]. Sharma, S., Gupta, P., Gupta, A., Kawish, S., Iqbal, Z., Vohora, D., & Kohli, K. (2023). Rapid Analytical Method Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Exemestane and Genistein with Specific Application in Lipid-Based Nanoformulations. ACS Omega, 8(28), 25101–25113. https://doi.org/10.1021/acsomega.3c01791
[20]. Thompson, M., Ellison, S.L.R , Wood, R. (2015). The IUPAC Technical report therefore presents the international harmonized protocol for the proficiency testing of analytical chemistry laboratories. Journal of Pure and Applied Chemistry, 78, 1, 145-196. https://doi.org/10.1515/pac-2013-0508
[21]. Tocher, D. R., Harvie, D. G., & Glencross, B. D. (2008). Fatty acid compositions of the major phospholipids of fish cells and tissues: LA and ALA in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) fed n−3 and n−6 polyunsaturated fatty acids. Lipids, 43(7), 647-657. https://doi.org/10.1007/s11745-008-3185-y
[22]. Zhang, Xia; Yap, Yee Gin; Wei, Da; Chen, Feng; Danchin, Antoine; Jiang, Hui. Single drop microextraction with ionic liquid and high-performance liquid chromatography coupled with quadrupole linear ion trap mass spectrometry for analysis of fatty acid profile in olive oils. Analytica Chimica Acta 2008, 716, 127-132. https://doi.org/10.1016/j.aca.2011.12.036