STABILITY-INDICATING REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF METHYLCOBALAMIN, ALPHA-LIPOIC ACID, PYRIDOXINE HCL, AND FOLIC ACID IN BULK AND COMBINED DOSAGE FORM
DOI:
https://doi.org/10.22159/ajpcr.2018.v11i12.26715Keywords:
Alpha-lipoic acid, Folic acid, Methylcobalamin, Pyridoxine hydrochloride and Reversed-phase high-performance liquid chromatography, ValidationAbstract
Objectives: The purpose of the research is to develop a simple, precise, economical, accurate, reproducible, and sensitive method for the estimation of methylcobalamin, alpha-lipoic acid, pyridoxine hydrochloride, and folic acid drug product by reversed-phase high-performance liquid chromatography (RP-HPLC) method.
Methods: New analytical method was developed for the estimation of methylcobalamin, alpha-lipoic acid, pyridoxine hydrochloride, and folic acid in drug product by RP-HPLC. The chromatographic separation was achieved on the Inertsil C18, 250 mm × 4.6 mm, 5 μm at ambient temperature. The separation achieved employing a mobile phase consists of buffer (added 5.05 g hexane-1-sulfonic acid is dissolved into 1000 mL of distilled water):acetonitrile in the ratio of 10:90% v/v. The flow rate was 1 mL/min and UV-visible spectrophotometer at 285 nm. The average retention time for methylcobalamin, alpha-lipoic acid, pyridoxine hydrochloride, and folic acid was found to be 3.5, 6.7, 8.5, and 9.3, respectively.
Results: The developed method was validated as per ICH guidelines. All validation parameters were within the acceptable ranges. The assay methods were found to be linear from 0 to 2130 μg/mL for methylcobalamin, 0 to 142.5 μg/mL for alpha-lipoic acid, 0–4.54 μg/mL for pyridoxine hydrochloride, and 0–2 μg/mL for folic acid. The correlation coefficient was 0.999 for all drugs, respectively. The mean percentage values for the developed method were found to be within the range of 98–100.6%. The developed method was also found to be robust.
Conclusion: It is concluded that developed method was accurate, precise, linear, reproducible, robust, and sensitive.
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References
McDowell LR. Vitamins in Animal and Human Nutrition. Available from: https://www.Booksgoogle.com. [Last retrieved on 2018 Jan 28].
Eisai Submits New Drug Application for Mecobalamin Ultra-High Dose Preparation as Treatment for Amyotrophic Lateral Sclerosis In Japan (PDF). Available from: https://www.Eisai.com. [Last retrieved on 2018 Jan 28].
World Health Organization. WHO Model Formulary 2008 (PDF). World Health Organization; 2009. p. 496.Available from: ???. [Last retrieved on 2016 Dec 08].
van Woerden CS, Groothoff JW, Wijburg FA, Annink C, Wanders RJ, Waterham HR, et al. Clinical implications of mutation analysis in primary hyperoxaluria Type 1. Kidney Int 2004;66:746-52.
Monico CG, Olson JB, Milliner DS. Implications of genotype and enzyme phenotype in pyridoxine response of patients with Type I primary hyperoxaluria. Am J Nephrol 2005;25:183-8.
Monico CG, Rossetti S, Olson JB, Milliner DS. Pyridoxine effect in Type I primary hyperoxaluria is associated with the most common mutant allele. Kidney Int 2005;67:1704-9.
Dietary supplement fact sheet. Folate. Health Information. Office of Dietary Supplements, US: National Institutes of Health; 2014. Available from: https://www.ods.od.nih.gov/factsheets/Folate-HealthProfessional.
Weinstein SJ, Hartman TJ, Stolzenberg-Solomon R, Pietinen P, Barrett MJ, Taylor PR, et al. Null association between prostate cancer and serum folate, Vitamin B(6), Vitamin B(12), and homocysteine. Cancer Epidemiol Biomarkers Prev 2003;12:1271-2.
Nawaz M. A new validated stability indicating RP-HPLC method for simultaneous estimation of pyridoxine hydrochloride and meclizine hydrochloride in pharmaceutical solid dosage forms. Chromatogr Res Int 2013;2013 Article ID: 747060. pages:7.
Shah R, Shah R. Stability indicating RP-HPLC method for simultaneous estimation of dosulepin hydrochloride and methylcobalamin in tablet dosage form. Int J App pharm 2017;9:69-75.
Available from: https://www.innovareacademics.in/journals/index.php/ ijpps/article/view/3369.
Available from: https://www.ijprd.com/ENT%20AND%20VALIDATION%20 OF%20ANALYTICAL.
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