DEVELOPMENT AND VALIDATION OF A GRADIENT HPLC METHOD FOR QUANTIFICATION OF EDETATE DISODIUM IN LYOPHILIZED INJECTABLE DRUG PRODUCT
Objective: The present study was aimed to validate a developed reversed phase gradient high-performance liquid chromatography method for the quantitative determination of Edetate Disodium in the lyophilized injectable drug product.
Methods: The amount of total Edetate disodium was analysed by HPLC assay using Edetate disodium USP as a reference standard. Injectable product was dissolved in acetone and Edetate disodium is separated out from API and then dissolved in water. Analysis was carried out using ferric chloride as a precolumn derivatizing reagent and YMC Pack ODS-A, 5 µm column with mobile phase as a mixture of tetrabutylammonium bromide buffer pH 2.8 and acetonitrile as the solvent, water used as diluent. The Edetate disodium quantified by U. V. wavelength at 254 nm.
Results: The method was suitably validated with respect to specificity, linearity, precision, accuracy and solution stability, using this method the average recovery from spike sample is 98.2%, with a relative standard deviation of<3%. The minimal quantifiable level was 1.5 µg/ml. The results show that the procedure is accurate, precise and reproducible.
Conclusion: In the present study an attempt has been made to develop and validate the analytical method for lyophilised injectable formulations and to generate the scientific database for formulation and evaluation of various lyophilised injectable containing Edetate disodium.
2. Hawken CM. Chelation therapy: an effective method for maintaining cardiovascular health, history of EDTA and chelation therapy. Texas: Woodland publishing; 1997.
3. Nema S, Brendel RJ. Excipients and their role in approved injectable products: current usage and future directions. PDA J Pharm Sci Technol 2011;65:287-332.
4. Domingo JL. Developmental toxicity of metal chelating agents. Reprod Toxicol 1998;12:400-510.
5. Saito K, Hasuo T, Nakano H. Nippon jozo kyokai zasshi determination and differentiation of ethylenediaminetetra-acetic acid (EDTA) and nitrilotriacetic acid (NTA) in fresh water. Water Res 1968;63:1193-7.
6. Laine P, Matilainen R. Simultaneous determination of DTPA, EDTA and NTA by UV spectrometry and HPLC. Anal Bioanal Chem 2005;382:1601-9.
7. Kaiser KLE. Removal of some chelators from aqueous solutions using polymeric ingredients. Water Res 1973;7:1465-73.
8. Nemocova I, Pesinova H, Suk V. Analysis of amino-polycarboxylates and organophosphonates. Microchem 1984; 30:27-32.
9. Nikolelis DP, Hadjiioannou TP. All-solid-state potentiometric sensors for ascorbic acid by using a screen-printed compatible solid contact. Anal Chim Acta 1978;97:111-20.
10. Qureshi SZ, Bansal R, Fresenius Z. Cation-exchange method for the detection of pyrrole derivatives. Anal Chem 1981;308:32.
11. Qureshi SZ, Bansal R. Analysis of aminopolycarboxylates and organophosphonates; 1984;12:42-4.
12. Blijenberg BG, Leijnse B. A simple method for the determination of EDTA in serum and Urine. Chim Acta 1969;26:577-9.
13. Huber CO, Tallant DR. Voltammetry at the lead dioxide electrode: anodic EDTA currents. J Electroanal Chem Interfacial Electrochem 1968;18:421-5.
14. Milwidsky BM. Determination and differentiation of ethylenediaminetetra-acetic acid (EDTA) and nitrilotriacetic acid (NTA). Soap Cosmet Chem Spec 1971;47:46, 49.
15. Slowinski EJ, Masterton WL, Wolsey WC. Chemical principles in the laboratory. WB Saunders: Philadelphia; 1969. p. 506-7.
16. Fitzgerald EA. Phenacyl esters of fatty acids via crown ether catalysts for enhanced ultraviolet detection in liquid chromatography. Anal Chem 1975;47:356-7.
17. Parkes DG, Caruso MG, Spradling JE. Determination of nitrilotriacetic acid in ethylenediaminetetraacetic acid disodium salt by reverse-phase ion-pair liquid chromatography. Anal Chem 1981;53:2154-6.
18. U. S. Pharmacopeia. 21st Rev, US. Pharrnacopeial Convention: Rockville, MD; 1985. p. 368-9.
19. Venezky DL, Rudzinski WE. Handbook of detergents, part c. Anal Chem 1984;56:315-7.
20. Yamaguchi A, Rajput AR, Ohzeki K, Kambara T. Determination of EDTA in vancomycin by liquid chromatography with absorbance ratioing for peak identification. Bull Chern Soc J 1983;56:2621-23.
21. Fitchett AW, Woodruff A. Analysis of aminopolycarboxylates and organophosphonates liq. Chromatogram HPLC Mag 1983;1:48-9.
22. Yamaguchi A, Ohzeki K, Kambara T, Fresenius’ Z. Determination of trace amounts of EDTA by use of a CuO-loaded cation exchanger. Anal Chem 1982;310:30-2.
23. Bauer J, Heathcote D, Krogh SJ. Quantitative determination of disodium Edetate in ophthalmic and contact lens care solution by reverse-phase high-performance chromatography. J Pharma Sci 1986;369:422-5.
24. www.drugbank.com [Last accessed on 10 Jan 2019]
25. www.sciencedirect.com [Last accessed on 10 Jan 2019]
26. www.pubmed.com [Last accessed on 10 Jan 2019]
27. www.mhra.gov.uk [Last accessed on 10 Jan 2019]
28. International conference on harmonization of technical requirements for registration of pharmaceuticals for human use. ICH Guideline Q2 (R1); 1995.
29. USP 41-NF 36 Online Table of Contents: Admissions; 2007.
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