Development of Validated Spectrophotometric and Chromatographic Methods for Simultaneous Estimation Taxifolin and Bergapten in Marketed Formulation
Main Article Content
Abstract
Background: Bergapten (BER) and taxifolin (TXN), a biomarkers which has many therapeutic activities such as anti-oxidant, anti-inflammatory, anti-microbial, and other pharmacological actions. This shows a new, simple, precise, and accurate High Performance Thin Layer Chromatography (HPTLC) and UV methods for estimating Taxifolin (TXN) and Bergapten (BER) as biomarkers in marketed formulations. Aim: The study determines simultaneous estimation and Q-analysis of UV-Vis spectrophotometry as well as to develop and validate method parameter in High Performance Thin Layer Chromatography (HPTLC). Objective: Application of developed method to quantify these biomarkers in commercial formulation. Materials and methods: The chemicals and reagents were purchased from Merck in India and were of analytical grade. The biomarkers were used of Tokyo Chemical Industry Co., Ltd. bergapten (98.0% w/w) and (+)-taxifolin (95.0%). The biomarkers were estimated and analysed with UV Probe 2.0 software and winCATS software relatively. Methods performed in UV-Vis spectrophotometry (i) Simultaneous estimation and (ii) Q-analysis whereas for HPTLC the biomarkers were simultaneously estimated on parameters of method development. Results: The method for UV-Vis spectroscopy were analysed with a LOD and LOQ of 0.5 and 1 respectively where the %RSD does not exceed more than 2%. The biomarkers were separated by chromatography on precoated aluminium plates as the stationary phase and where optimized by using mobile phase such as toluene:ethyl acetate:formic acid (4:6:0.1 v/v/v) at 306 nm densitometry, and where validated at separate zones. Taxifolin and Bergapten were resolved satisfactorily, with Rf values of 0.64 and 0.82, respectively, in accordance with ICH guidelines. Conclusion: The newly designed UV-Vis Spectroscopy and HPTLC technique has improved specificity, precision and accuracy. The quality of biomarkers containing a flavonoid and furanocoumarins were being validated and quantify these biomarkers in commercial formulation.
Article Details
References
Pew JC. A flavonone from Douglas-fir heartwood. J Am Chem Soc.1948;70(9):3031- Sawant RL, Hadawale SD, Dhikale GK, Bansode CA, Tajane PS. Spectrophotometric methods for simultaneous estimation of rabeprazole sodium and aceclofenac from the combined capsule dosage form. Pharm Methods. 2011;2(3):193–7.doi: 10.4103/2229-4708.903624. doi: 10.1021/ja01189a059, PMID 18882535.
Wallace SN, Carrier DJ, Clausen EC. Batch solvent extraction of flavanolignans from milk thistle (Silybum marianum L. Gaertner). Phytochem Anal. 2005;16(1):7-16. doi: 10.1002/pca.803. PMID 15688950.
Slimestad R, Fossen T, Vågen IM. Onions: A source of unique dietary flavonoids.J Agric Food Chem. 2007;55(25):10067-80. doi: 10.1021/jf0712503, PMID 17997520.
Kiehlmann E, Li EPM. Isomerization of dihydroquercetin. J Nat Prod.1995;58(3):450-5. doi: 10.1021/np50117a018 R. Slimestad R, Fossen T, Vågen IM. Onions: A source of unique dietary flavonoids. J Agric Food Chem.2007;55(25):10067-80, doi: 10.1021/jf0712503, PMID 17997520.
Rohdewald P. A review of the French maritime pine bark extract (Pycnogenol),a herbal medication with a diverse clinical pharmacology. Int J Clin Pharmacol Ther. 2002;40(4):158-68. doi: 10.5414/cpp40158, PMID 11996210.
M. C. Chavan, Navratne AR, Patil RB, Vanjari SS. Development and Validation of Stability Indicating High Performance Thin Layer Chromatography Method for Analysis of Bergapten. Tathawade; Maharashtra, India, J. Pharm. Sci. & Res. Vol. 11(9), 2019, 3237-3242
Srinivas B, Ashraf S, Vivek N, Rajashree H. Development and validation of bivariate UV-visible spectroscopic method for simultaneous estimation of curcumin and piperine in their combined nanoparticulate system. J Appl Pharm Sci. 2021;doi: 10.7324/japs.2021.110509
Gomathi D, Kalaiselvi M, Ravikumar G, Sophia D, Gopalakrishnan VK, Uma C. Secondary metabolite credentials of Evolvulus alsinoides by high performance thin layer chromatography (HPTLC). J Biomed Res. 2012;26(4):295–302.doi: 10.7555/JBR.26.20110128
Sawant RL, Hadawale SD, Dhikale GK, Bansode CA, Tajane PS. Spectrophotometric methods for simultaneous estimation of rabeprazole sodium and aceclofenac from the combined capsule dosage form. Pharm Methods. 2011;2(3):193–7.doi: 10.4103/2229-4708.90362
Bhaskar KL, Lakshmi DS, Sumalatha G, Suji G, Kumar KAT. Q-analysis and simultaneous equation method for estimation of domperidone and naproxen by UV spectrophotometry in bulk and tablet dosage form. Res J Pharm Technol. 2020;13(12):6050–4.doi: 10.5958/0974-360x.2020.01054.9
A. K. S. Rawat, A. P. Singh, D. P. Singh, M. M. Pandey, R. Govindarajan, and Sharad Srivastava. Separation and Identification of Furocoumarin in Fruits of Heracleum candicans DC. Hindawi Publishing Corporation; Volume 2013, Article ID 915762, 4 pages, 2013.doi: 10.1155/2013/915762
Srivastava, M. (2010). An Overview of HPTLC: A Modern Analytical Technique with Excellent Potential for Automation, Optimization, Hyphenation, and Multidimensional Applications. High-Performance Thin-Layer Chromatography (HPTLC), 3–24. doi:10.1007/978-3-642-14025-9_1
ICH guidelines, Q1A(R2): Stability Testing of New Drug Substances and Products. (revision 2). International Conference on Harmonization. 2003.
Urška Jug, Irena Vovk, Vesna Glavnik, Damjan Makuc, Katerina Naumoska. Off-line multidimensional high performance thin-layer chromatography for fractionation of Japanese knotweed rhizome bark extract and isolation of flavan-3-ols, proanthocyanidins and anthraquinones. Journal of Chromatography A 1637 (2021) 461802; 2020.
Sampathkumar S, Ramakrishnan N. Chromatographic fingerprint analysis of Naringi crenulata by HPTLC Technique. Asian Pac J Trop Biomed 2011(1S); 195-8.
Yogesh P Pancham, Girish B and Shailendra Suryawanshi Sanjay, UV-Spectrophotometric method for quantification of ascorbic acid in bulk powder, The Pharma Innovation Journal 2020; 9(5): 05-08
Glassco J. Analytical Method Development and Validation. LLS Health CDMO. Lubrizol CDMO.2019; [cited 2023 May 12]. Available from: https://lubrizolcdmo.com/technical-briefs/analytical-method-development-and-validation/
Le Borgne E, Cicchetti E, Bertrand T. HPTLC methods for qualitative and quantitative analysis of selected furocoumarins in essential oils. Flavour Fragr J. 2017;32(5):330–9. http://dx.doi.org/10.1002/ffj.3394
Mignot B, Guillaume Y, Makki S, Murret E, Cavalli E, Truong TT, et al. High-performance thin-layer chromatographic determination of 5-methoxypsoralen in serum from patients. J Chromatogr. 1997;700(1–2):283–5.doi: 10.1016/s0378-4347(97)00229-6
Sawant RL, Hadawale SD, Dhikale GK, Bansode CA, Tajane PS. Spectrophotometric methods for simultaneous estimation of rabeprazole sodium and aceclofenac from the combined capsule dosage form. Pharm Methods 2011;2(3):193–7.doi: 10.4103/2229-4708.90362
Pancham YP, Sanjay SS. UV-Spectrophotometric method for quantification of ascorbic acid in bulk powder [Internet]. Thepharmajournal.com. [cited 2023 May 12]. Available from: https://www.thepharmajournal.com/archives/2020/vol9issue5/PartA/8-6-15-737.pdf
Jinxiu Hou, Mingyang Hu, et al, Dietary Taxifolin Protects Against dextran Sulfate Sodium-Induced Colitis via NF-κB Signaling, Enhancing Intestinal Barrier and Modulating Gut Microbiota, Frontiers in Immunology, 2021, Volume 11. doi: 10.3389/fimmu.2020.631809
Sunil C, Xu B. An insight into the health-promoting effects of taxifolin(dihydoquercetin).Phytochemistry.2019;166:112066.doi10.1016/j.phytochem.2019.112066
Melough, M. M., Cho, E., Chun, O. K., Food Chem. Toxicol. 2018, 113, 99-107
Hung, W. L., Suh, J. H., Wang, Y., Food Drug Anal. 2017, 25, 71-83
Kubrak, T., Podgorski, R., Stompor, M., Eur. J. Clin. Exp. Med. 2017, 15, 169-175
Moura FCS. Taxifolin stability: In silico prediction and in vitro degradation with HPLC-UV/UPLCeESI-MS monitoring. Journal of Pharmaceutical Analysis. 2020 Jun 29;11:232–40.
UV-Vis spectroscopy: Principle, instrumentation, and applications [Internet]. PSIBERG. 2021 [cited 2023 May 12]. Available from: https://psiberg.com/uv-vis-spectroscopy/