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Development of a New Analytical Method for Determination of Related Components in Nateglinide


An isocratic reverse phase liquid chromatographic (RP-LC) assay method has been developed for the quantitative determination of nateglinide and its related components namely imp-1 and imp-2 in bulk drug and in pharmaceutical dosage form, used for the treatment of type II diabetes mellitus. The developed method is stability indicating and also can be used for stability testing. The chromatographic separation was achieved on C-8, 150 × 4.6 mm, 3.5 μm stationary phase. The LC method employs solution A as mobile phase. Solution A contains a mixture of phosphate buffer pH 3.0: acetonitrile (50:50 v/v). The flow rate was 1.0 mL min−1 and the detection wavelength was 210 nm. In the developed LC method the resolution between nateglinide and its potential impurities namely imp-1 and imp-2 was found to be greater than 5.0. The drug was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. Considerable degradation was found to occur in acid medium, alkaline medium and oxidative stress conditions. The stress samples were assayed against a qualified reference standard and the mass balance was found close to 99.2%. The developed RP-LC method was validated with respect to linearity, accuracy, precision and robustness.

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  1. Fujitani S, Okazaki K, Yada T (1997) Br J Pharmacol 120:1191–1198

    Article  CAS  Google Scholar 

  2. Karara AH, Dunning BE, McLeod JF (1999) J Clin Pharmacol 39:172–179

    Article  CAS  Google Scholar 

  3. Dunn CJ, Faulds D (2000) Drugs 60:607–615

    Article  CAS  Google Scholar 

  4. Fujitani S, Yada T (1994) Endocrinology 134:1395–1400

    Article  CAS  Google Scholar 

  5. Yoshikazu N, Hiroyui A, Makoto I, Kazutoshi T, Kazuo H, Shoj T (1999) Yakuri to Chiryo 27:1177–1203

    Google Scholar 

  6. Keilson L, Mather S, Walter YH, Subramanian S, Mcleod JF (2000) J Clin Endocrinol .Metab 85:1081–1086

    Article  CAS  Google Scholar 

  7. Sankalia JM, Sankalia MG, Sutariya VB, Mashru RC (2007) J Pharm Biomed Anal 44:196–204

    Article  CAS  Google Scholar 

  8. Hongyuan Yan, Gengliang Yang, Fengxia Qiao, Yi Chen (2004) J Pharm Biomed Anal 36:169–174

  9. Bauer S, Störmer E, Kirchheiner J, Michael C, Brockmöller J, Roots I (2003) J Pharm Biomed Anal 31:551–555

    Article  CAS  Google Scholar 

  10. Ono I, Matsuda K, Kanno S (1997) J Chromatogr B 692:397–404

    Article  CAS  Google Scholar 

  11. Ono I, Matsuda K, Kanno S (1996) J Chromatogr B Biomed Sci Appl 678:384–387

    Article  CAS  Google Scholar 

  12. Malli D, Gikas E, Vavagiannis A, Kazanis M, Daniilides K, Gennimata D, Panderi I (2007) Anal Chim Acta 599:143–150

    Article  CAS  Google Scholar 

  13. ICH Stability Testing of New Drug Substances and Products Q1A (R2) (2003) International Conference on Harmonization, IFPMA, Geneva

  14. Singh S, Bakshi M (2000) Pharm Tech On-line 24:1–14

    Google Scholar 

  15. ICH, Photo Stability Testing of New Drug Substances and Products Q1B (1996) International Conference on Harmonization, IFPMA, Geneva

  16. Bakshi M, Singh S, Pharm J (2002) Biomed Anal 28:1011–1040

    Article  CAS  Google Scholar 

  17. Singh S, Singh B, Bahuguna R, Wadhwa L, Saxena R (2006) J Pharm Biomed Anal 41:1037–1040

    Article  CAS  Google Scholar 

  18. Carstensen JT, Rhodes CT (eds) (2000) Drug stability principles and practices, 3rd edn

  19. ICH Guidelines on Validation of Analytical Procedures: Text and Methodology Q2 (R1) (1995): FDA: Published in the Federal Register, vol 60, p 11260, March 1, 1995

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The authors wish to thank the management of Dr. Reddys Laboratories Ltd for supporting this work.

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Correspondence to A. Madhavi.

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Madhavi, A., Reddy, G.S., Suryanarayana, M.V. et al. Development of a New Analytical Method for Determination of Related Components in Nateglinide. Chroma 67, 639–645 (2008).

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  • Column liquid chromatography
  • Forced degradation
  • Validation and stability indicating
  • Nateglinide