Year 2024,
Volume: 4 Issue: 3, 77 - 80, 03.07.2024
Merve Arslan
Durişehvar Ünal
References
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- 2. Orobia AJM, Saa J, Lorenzo AO, Herreras JM. Combination of hyaluronic acid, carmellose, and osmoprotectants for the treatment of dry eye disease. Clin. Ophthalmol. 2018;Volume 12:453-461. [CrossRef]
- 3. Liao YH, Jones SA, Forbes B, Martin GP, Brown MB. Hyaluronan: Pharmaceutical Characterization and Drug Delivery. Drug Delivery. 2005;12(6):327-342. [CrossRef]
- 4. Hanlan J, Skoog DA, West DM. Principles of Instrumental Analysis. Stud. Conserv. 1973;18(1):45. [CrossRef]
- 5. Gässler N, Reißner C, Janzen N, Kähnert H, Kleesiek K. A High Performance Liquid Chromatography Method for the Determination of Glycosaminoglycans in Human Blood. Clin. Chem. Lab. Med. 1993;31(8). [CrossRef]
- 6. Ruckmani K, Shaikh SZ, Khalil P, Muneera, Thusleem OA. Determination of sodium hyaluronate in pharmaceutical formulations by HPLC–UV. J. Pharm. 2013;3(5):324-329. [CrossRef]
- 7. Aaltonen K, Niemelä T, Sankari S, Tulamo RM. Determination of the unsaturated disaccharides of hyaluronic acid in equine synovial fluid by high-performance liquid chromatography and fluorescence detection. Acta Vet. Scand. 2015;57(1):12. [CrossRef]
- 8. Grøndahl F, Tveit H, Akslen-Hoel LK, Prydz K. Easy HPLC-based separ-ation and quantitation of chondroitin sulphate and hyaluronan disaccharidesafter chondroitinase ABC treatment. Carbohydr. Res. 2011;346:50–57. [CrossRef]
- 9. Al-Sibani M, Al-Harrasi A, Neubert RHH. Evaluation of in-vitro degradation rate of hyaluronic acid-based hydrogel cross-linked with 1,4-butanediol diglycidyl ether (BDDE) using RP-HPLC and UV–vis spectroscopy. J. Drug Deliv. Sci. Tec. 2015;29:24-30. [CrossRef]
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- 16. Xu X, Wen C, Ai C, et al. Quantitative Analysis of Acidic Polysaccharides Using Hydrophilic Interaction Chromatography and Mass Spectrometry after Acid Hydrolysis. Curr. Pharm. Anal. 2018;14(5):443-449. [CrossRef]
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Determination of Hyaluronic Acid in Ophthalmic Solution by HPLC–UV and Stability Analysis
Year 2024,
Volume: 4 Issue: 3, 77 - 80, 03.07.2024
Merve Arslan
Durişehvar Ünal
Abstract
Objective: Hyaluronic acid (HA) is a naturally occurring, biocompatible polysaccharide with unique viscoelastic and hygroscopic properties. Its role as a natural lubricant and its excellent water-retaining properties make it well-suited for use in ophthalmic products. The objective of this study is determination of sodium hyaluronate in ophthalmic solution liquid chromatography (HPLC) method with UV detection and, stability of the product.
Methods: The analysis was carried out using a polymer column (PolySep-GFC- P5000) with a mobile phase consisting of (dH2O: 20 mM Phosphate Buffer pH:6.5), (95:5, v:v) at a flow rate of 1.2 mL/min. UV detection was set 205 nm.
Results: Calibration curve was linear over the concentration range of 80%-120% concentration of the ophthalmic product. The back calculated concentrations of the calibration standards were within ±15% of the nominal value (±20% for LOQ). The stability of ophtalmic solution was found to be 92.5% at the end of the days at room tempereture.
Conclusion: The developed method is fast and reliable for HA analysis in ophthalmic products. Accordingt to stability study, these products must be stored at +4 C during usage and storage.
Ethical Statement
Ethical approval and informed consent are not required in our study as no research was conducted on human or animal specimens.
Supporting Institution
The authors declared that this study has received no financial support.
References
- 1. Fraser JRE, Laurent TC, Laurent UBG. Hyaluronan: its nature, distribution, functions and turnover. J. Intern. Med. 1997;242(1):27-33. [CrossRef]
- 2. Orobia AJM, Saa J, Lorenzo AO, Herreras JM. Combination of hyaluronic acid, carmellose, and osmoprotectants for the treatment of dry eye disease. Clin. Ophthalmol. 2018;Volume 12:453-461. [CrossRef]
- 3. Liao YH, Jones SA, Forbes B, Martin GP, Brown MB. Hyaluronan: Pharmaceutical Characterization and Drug Delivery. Drug Delivery. 2005;12(6):327-342. [CrossRef]
- 4. Hanlan J, Skoog DA, West DM. Principles of Instrumental Analysis. Stud. Conserv. 1973;18(1):45. [CrossRef]
- 5. Gässler N, Reißner C, Janzen N, Kähnert H, Kleesiek K. A High Performance Liquid Chromatography Method for the Determination of Glycosaminoglycans in Human Blood. Clin. Chem. Lab. Med. 1993;31(8). [CrossRef]
- 6. Ruckmani K, Shaikh SZ, Khalil P, Muneera, Thusleem OA. Determination of sodium hyaluronate in pharmaceutical formulations by HPLC–UV. J. Pharm. 2013;3(5):324-329. [CrossRef]
- 7. Aaltonen K, Niemelä T, Sankari S, Tulamo RM. Determination of the unsaturated disaccharides of hyaluronic acid in equine synovial fluid by high-performance liquid chromatography and fluorescence detection. Acta Vet. Scand. 2015;57(1):12. [CrossRef]
- 8. Grøndahl F, Tveit H, Akslen-Hoel LK, Prydz K. Easy HPLC-based separ-ation and quantitation of chondroitin sulphate and hyaluronan disaccharidesafter chondroitinase ABC treatment. Carbohydr. Res. 2011;346:50–57. [CrossRef]
- 9. Al-Sibani M, Al-Harrasi A, Neubert RHH. Evaluation of in-vitro degradation rate of hyaluronic acid-based hydrogel cross-linked with 1,4-butanediol diglycidyl ether (BDDE) using RP-HPLC and UV–vis spectroscopy. J. Drug Deliv. Sci. Tec. 2015;29:24-30. [CrossRef]
- 10. Hjerpe A. Liquid-chromatographic determination of hyaluronic acid in pleural and ascitic fluids. Clin. Chem. 1986;32:952-956. [CrossRef]
- 11. Alkrad JA, Merstani Y, Neubert RH. New approaches for quantifying hyaluronic acid in pharmaceutical semisolid formulations using HPLC and CZE. J. Pharm. Biomed. Anal. 2002;30:913–919. [CrossRef]
- 12. Nakano T, Nakano K, Sim JS. A Simple Rapid Method To Estimate Hyaluronic Acid Concentrations in Rooster Comb and Wattle Using Cellulose Acetate Electrophoresis. J. Agric. Food Chem. 1994;42(12):2766-2768. [CrossRef]
- 13. Vigliano M, Bianchera A, Bettini R, Elviri L. Determination of Hyaluronic Acid in a Chitosan-Based Formulation by RP C18 and HILIC LC–ESI-MS: an Evaluation of Matrix Effect. Chromatographia. 2013;76(23-24):1761-1766. [CrossRef]
- 14. Song S, Yu Q, Zhang B, et al. Glycoconj. J. 2017; 34: 625–632. [CrossRef]
- 15. Kühn AV, Raith K, Sauerland V, Neubert RHH. Quantification of hyaluronic acid fragments in pharmaceutical formulations using LC–ESI–MS. J. Pharm. Biomed. Anal. 2003;30(5):1531-1537. [CrossRef]
- 16. Xu X, Wen C, Ai C, et al. Quantitative Analysis of Acidic Polysaccharides Using Hydrophilic Interaction Chromatography and Mass Spectrometry after Acid Hydrolysis. Curr. Pharm. Anal. 2018;14(5):443-449. [CrossRef]
- 17. Karlsson N, Schulz B, Packer N, Whitelock J. Use of graphitised carbon negative ion LC–MS to analyse enzymatically digested glycosaminoglycans. J. Chromatogr. B. 2005;824(1-2):139-147. [CrossRef]
- 18. Song JM, Im JH, Kang JH, Kang DJ. A simple method for hyaluronic acid quantification in culture broth. Carbohydr. Polym. 2009;78(3):633-634. [CrossRef]
- 19. Oueslati N, Leblanc P, Harscoat-Schiavo C, et al. CTAB turbidimetric method for assaying hyaluronic acid in complex environments and under cross-linked form. Carbohydr. Polym. 2014;112:102-108. [CrossRef]
- 20. Akiyama H, Shidawara S, Mada A, et al. Chemiluminescence high-performance liquid chromatography for the determination of hyaluronic acid, chondroitin sulphate and dermatan sulphate. J. Chromatogr. B Biomed. Appl. 1992;579(2):203-207. [CrossRef]
- 21. Tokita Y, Okamoto A. Hydrolytic degradation of hyaluronic acid. Polym. Degrad. Stab. 1995;48(2):269-273. [CrossRef]
- 22. Maleki A, Kjøniksen A, Nyström B. Effect of pH on the Behavior of Hyaluronic Acid in Dilute and Semidilute Aqueous Solutions. Macromol. Symp. 2008;274(1):131-140. [CrossRef]
- 23. FDA (Center for Drug Evaluation and Research, U.S. Food and Drug Administration). Reviewer Guidance, Validation of Chromatographic Methods; 1994.