A NOVEL RP-HPLC GRADIENT ELUTION TECHNIQUE FOR BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATING GALLIC ACID IN WISTAR RAT PLASMA
Keywords:Gallic acid, Rat plasma, RP-HPLC, Gradient elution, Bioanalytical method, Validation
Objective: Present study aimed to develop and validate a novel, unique, simple, quick, cost-effective, sensitive, specific, accurate, precise, rugged, and robust bioanalytical method for the quantification of gallic acid in rat plasma by reverse phase high-performance liquid chromatography (RP-HPLC) using gradient elution technique.
Methods: The stationary phase was a Zorbax SB C18 5 µ (4.6*150) mm column, with the mobile phase being water with 0.1 percent formic acid (A): acetonitrile (ACN) with 0.08 percent formic acid (B). Gradient chromatographic method was used throughout this experiment from the point of view of the estimation of gallic acid from herbal formulations when present along with other phytoconstituents. So at the gradient method, all the present phytoconstituents has cleared off from the column and no any strongly adsorption of phytoconstituents occurred. The experiment was carried out at a flow rate of 1.0 ml/min at 30 °C utilising PDA detectors at 271 nm. The proposed method was validated for different parameters.
Results: The approach was found to be linear in the concentration range of 0.5-100 µg/ml, with a r2 of 0.9998. There was not observed any interference of co-eluting peaks of endogenous compounds from the biological matrix at the same retention time (Rt) of gallic acid. The RSD (%) of intra and interday precision was found to be within acceptable limit. The overall % mean recovery was found to be 99.97%. LOD and LOQ were found to be 0.1 and 0.5 μg/ml, respectively. In terms of fluctuation in essential parameters and operating settings, the devised bioanalytical approach was shown to be rugged and resilient. Short-term, long-term, autosampler, bench-top, and freeze-thaw stability experiments revealed that gallic acid is stable.
Conclusion: The developed method described in this report was found to be well within an acceptable range. Hence, in the future, this method can be used successfully for the estimation of gallic acid alone or in combination with another analyte or marker present in bulk or an extract containing various phytoconstituents in pharmacokinetic, bioequivalence, and therapeutic drug monitoring studies in clinical laboratories.
Naaz H, Srikanth P, Rudrapal M, Sarwa KK. Development and validation of UV spectrophotometric and RP-HPLC methods for the estimation of gallic acid in the herbal formulation of amalaki. Asian J Chem. 2020;32(10):2469-74. doi: 10.14233/ajchem.2020.22737.
Gao J, Hu J, Hu D, Yang X. A role of gallic acid in oxidative damage diseases: A comprehensive review. Nat Prod Commun. 2019 Aug;14(8). doi: 10.1177/1934578X19874174.
Brewer MS. Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Compr Rev Food Sci Food Saf. 2011 Jun 14;10(4):221-47. doi: 10.1111/j.1541-4337.2011.00156.x.
Kosuru RY, Roy A, Das SK, Bera S. Gallic acid and gallates in human health and disease: do mitochondria hold the key to success? Mol Nutr Food Res. 2018 Jan;62(1). doi: 10.1002/mnfr.201700699, PMID 29178387.
Su TR, Lin JJ, Tsai CC, Huang TK, Yang ZY, Wu MO. Inhibition of melanogenesis by gallic acid: possible involvement of the PI3K/Akt, MEK/ERK and Wnt/β-catenin signaling pathways in B16F10 cells. Int J Mol Sci. 2013 Oct 14;14(10):20443-58. doi: 10.3390/ijms141020443, PMID 24129178.
Sawa T, Nakao M, Akaike T, Ono K, Maeda H. Alkylperoxyl radical-scavenging activity of various flavonoids and other phenolic compounds: implications for the anti-tumor-promoter effect of vegetables. J Agric Food Chem. 1999 Feb;47(2):397-402. doi: 10.1021/jf980765e, PMID 10563906.
Shao D, Li J, Tang R, Liu L, Shi J. Inhibition of gallic acid on the growth and biofilm formation of escherichia coli and streptococcus mutans. J Food Sci. 2015 Jun;80(6): M1299-305. doi: 10.1111/1750-3841.12902, PMID 25974286.
Sorrentino E, Succi M, Tipaldi L, Pannella G, Maiuro L, Sturchio M. Antimicrobial activity of gallic acid against food-related pseudomonas strains and its use as biocontrol tool to improve the shelf life of fresh black truffles. Int J Food Microbiol. 2018 Feb 2;266:183-9. doi: 10.1016/j.ijfoodmicro.2017.11.026. PMID 29227905.
Oh E, Jeon B. Synergistic anti-campylobacter jejuni activity of fluoroquinolone and macrolide antibiotics with phenolic compounds. Front Microbiol. 2015 Oct 13;6:1129. doi: 10.3389/fmicb.2015.01129, PMID 26528273.
Fu R, Zhang Y, Peng T, Guo Y, Chen F. Phenolic composition and effects on allergic contact dermatitis of phenolic extracts Sapium sebiferum (L.) Roxb. leaves. J Ethnopharmacol. 2015 Mar 13;162:176-80. doi: 10.1016/j.jep.2014.12.072, PMID 25576898. jep.2014.12.072.
Korani MS, Farbood Y, Sarkaki A, Fathi Moghaddam H, Taghi Mansouri M. Protective effects of gallic acid against the chronic cerebral hypoperfusion-induced cognitive deficit and brain oxidative damage in rats. Eur J Pharmacol. 2014 Jun 15;733:62-7. doi: 10.1016/j.ejphar.2014.03.044, PMID 24726557.
Zakaria F, Wan Ishak WR, Wan Ahmad WAN, Safuan S, Tengku Ismail TA. Hypoglycaemic and protective effects of Benincasa hispida aqueous extract in streptozotocin-induced diabetic rats. Sains Malays. 2022 Mar 31;51(3):783-93. doi: 10.17576/jsm-2022-5103-12.
Kshirsagar RR, Vaidya SA, Jain V. Development and validation of a novel RP-HPLC method for the simultaneous quantification of ascorbic acid, gallic acid, ferulic acid, piperine, and thymol in a polyherbal formulation. Indian J Nat Prod Resour. 2020. doi: 10.56042/ijnpr.v11i4.30002.
Fernandes FHA, Batista RSdA, de Medeiros FD, Santos FS, Medeiros ACD. Development of a rapid and simple HPLC-UV method for determination of gallic acid in schinopsis brasiliensis. Revista Brasileira de Farmacognosia. 2015;25(3):208-11. doi: 10.1016/j.bjp.2015.05.006.
Al-Rimawi F, Odeh I. Development and validation of an HPLC-UV method for determination of eight phenolic compounds in date palms. J AOAC Int. 2015 Sep-Oct;98(5):1335-9. doi: 10.5740/jaoacint.15-010, PMID 26525252.
Khatri S. Bioanalytical method development and validation for the estimation of levocetirizine in blood plasma by using Rp-hplc. J Drug Delivery Ther. 2018 Oct 1;8(5-s):288-92. doi: 10.22270/jddt.v8i5-s.1977.
Babu R, Rao A, Rao JV. Bioanalytical method development and validation for simultaneous estimation of paracetamol and cefixime by using RP-HPLC in rabbit plasma. Orient J Chem. 2016 Mar 25;32(1):701-7. doi: 10.13005/ojc/320178.
Satyavert, Gupta S, Nair AB, Attimarad M. Development and validation of bioanalytical method for the determination of hydrazinocurcumin in rat plasma and organs by HPLC-UV. J Chromatogr B Analyt Technol Biomed Life Sci. 2020;1156:122310. doi: 10.1016/j.jchromb.2020.122310. PMID 32835908.
Bhandari R, Kuhad A, Paliwal JK, Kuhad A. Development of a new, sensitive, and robust analytical and bio-analytical RP-HPLC method for in vitro and in vivo quantification of naringenin in polymeric nanocarriers. J Anal Sci Technol. 2019 Mar 1;10(1). doi: 10.1186/s40543-019-0169-1.
Reddy BKK, Sekhar KBC, Mohan CK. Bioanalytical method development and validation for the simultaneous determination of vildagliptin and telmisartan in rabbit plasma using RP-HPLC. J Pharm Res Int. 2021 Feb 13:76-86. doi: 10.9734/jpri/2021/v33i131141.
GA, KB, Prasad K. Development and validation of bioanalytical hplc method for simultaneous estimation of cilnidipine and nebivolol in human plasma. Int J Pharm Pharm Sci. 2017 Oct 2;9(10):253. doi: 10.22159/ijpps.2017v9i10.20237.
Ajitha A, Sujatha K, Abbulu K. Bioanalytical method development and validation of garenoxacin mesylate in human plasma by RP-HPLC. International Journal of Research in Pharmaceutical Sciences 2019;10(2):1314-20. doi: 10.26452/ijrps.v10i2.534.
Moni F, Sharmin S, Rony SR, Afroz F, Akhter S, Sohrab MdH. Bioanalytical method validation of Esomeprazole by high-performance liquid chromatography with PDA detection. ACHROM. 2021 Mar 19;33(2):120-6. doi: 10.1556/ 1326.2020.00769.
Kanala K, Hwisa TN, Chandu BR, Katakam P, Khagga M, Challa BR, Khagga B. Bioanalytical method development and validation of milnacipran in rat plasma by LC-MS/MS detection and its application to a pharmacokinetic study. J Pharm Anal. 2013 Dec;3(6):481-8. doi: 10.1016/j. jpha.2013.03.009.
D’cruz D, Babu A, Joshy E. Bioanalytical method development and validation of ticagrelor by RP-HPLC. Int J App Pharm. 2017 May 1;9(3):51. doi: 10.22159/ijap.2017v9i3.17452.
Bhowmick M, Bhowmick P, Sengodan T, Thangavel S. Development and validation of bioanalytical Rp hplc method for the estimation of metoprolol tartrate in rabbit plasma after transdermal and oral administration: application in pharmacokinetic studies. J Drug Delivery Ther. 2015 Jul 15;5(4). doi: 10.22270/jddt.v5i4.1118.
Bhalme VM, Jumade PP, Bawankar RD, Wanjari DS, Mundhada DR. Method development, validation and stability indicating studies for simultaneous estimation of anti-hypertensive drugs from the pharmaceutical formulation by RP-HPLC. J Drug Delivery Ther. 2020 Nov 30;10(6):120-32. doi: 10.22270/jddt.v10i6.4571.
Kitzman D, Cheng KJ, Fleckenstein L. HPLC assay for albendazole and metabolites in human plasma for clinical pharmacokinetic studies. J Pharm Biomed Anal. 2002 Oct;30(3):801-13. doi: 10.1016/s0731-7085(02)00382-5, PMID 12367706.
Gültekin Y, Ozturk N, Filazi A, Deniz A, Korkmaz C, Pezİk E. Simultaneous determination of albendazole sulfoxide and praziquantel from PLGA nanoparticles and validation of new HPLC method. JRP 2022;26(6):1608-18. doi: 10.29228/jrp.252.
Mujewar IN, Bhusnure OG, Jagtap SR, Gholve SB, Giram PS, Savangikar AB. A review on bioanalytical method development and various validation stages involved in method development using RP-HPLC. J Drug Delivery Ther. 2019 Aug 25;9(4-s):789-95. doi: 10.22270/jddt.v9i4-s.3422.
Kumar S, Debnath M, Rao J, Sankar D. A new bioanalytical method development and validation for simultaneous estimation of esomeprazole and naproxen in human plasma by using RP-HPLC. Br J Pharm Res. 2014 Jan 10;4(19):2312-27. doi: 10.9734/BJPR/2014/10918.
Kumar SA, Debnath M, Rao JV, Sankar DG. New validated stability-indicating Rp-HPLC method for simultaneous estimation of atorvastatin and ezetimibe in human plasma by using PDA detector. Adv Pharm Bull. 2015 Sep;5(3):385-91. doi: 10.15171/apb.2015.053, PMID 26504761.
Ramesh D, Habibuddin M. Development and validation of Rp-hplc method for the determination of alvimopan in rat plasma. Int J Pharm Pharm Sci. 2018 Oct 1;10(10):124. doi: 10.22159/ijpps.2018v10i10.29001.
Gurav P, Damle M. Bioanalytical method for estimation of teriflunomidein human plasma. Int J Pharm Pharm Sci. 2022 Sep 1:19-23. doi: 10.22159/ijpps.2022v14i9.45151.
Sellappan M, Devakumar D. Development and validation of rp-hplc method for the estimation of escitalopram oxalate and flupentixol dihydrochloride in combined dosage form and plasma. Int J Pharm Pharm Sci. 2021 Feb 1:61-6. doi: 10.22159/ijpps.2021v13i2.30158.
Kondeti HP, Sankar Dannana G. Development and validation of a stability-indicating rp-hplc method for the estimation of metformin, saxagliptin, and dapagliflozin. Asian J Pharm Clin Res. 2022 Feb 4:72-7. doi: 10.22159/ajpcr.2022.v15i3.42117.
AS, Prasad Panigrahy U. Stability indicating method development and validation of fimasartan by reverse-phase high-performance liquid chromatography in bulk and pharmaceutical dosage form. Asian J Pharm Clin Res. 2021 Feb 7:138-46. doi: 10.22159/ajpcr.2021.v14i2.39919.
Shah SK, Dey S, De S. Simultaneous determination of atorvastatin and atenolol in rabbit plasma by RP-HPLC method and its application in pharmacokinetic study. Int J Curr Pharm Sci 2022;14:72-8. doi: 10.22159/ijcpr.2022v14i2.1968.
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