Fast and selective HPLC-DAD method for determination of pholcodine and related substances


  • Ana Petkovska Pharmaceutical Quality Control, Alkaloid AD Skopje
  • Hristina Babunovska Pharmaceutical Quality Control, Alkaloid AD Skopje
  • Marina Stefova Institute of Chemistry, Faculty of Natural Sciences and Mathematics, Ss. Cyril and Methodius University, Skopje



pholcodine, impurities, validation, HPLC-DAD, opium alkaloids


Quality control of pharmaceuticals requires development of fast, efficient and reliable methods for determination of active compounds as well as known and very often unknown impurities within defined concentration ranges. In this work, a simple and rapid HPLC-UV-DAD method for identification and quantification of pholcodine process related impurities and some degradation products was developed and validated. Pholcodine and its five structural analogues such as morphine, codeine, thebaine, oripavine, and papaverine were separated in less than 10 minutes using reversed phase LiChrospher C-8 column. For optimal chromatographic performance with reproducible retention times, gradient elution with 2% ammonium hydroxide in water and acetonitrile was used. The method was validated by establishing its selectivity, specifity, sensitivity, linearity, intra- and inter-day precision and robustness. All tested parameters confirmed that the method is suitable for determination of pholcodine and its five impurities in pharmaceutical drug samples. The results obtained from real sample analysis give support to the suitability of the proposed method for the purpose of quality control.


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How to Cite

Petkovska, A., Babunovska, H., & Stefova, M. (2011). Fast and selective HPLC-DAD method for determination of pholcodine and related substances. Macedonian Journal of Chemistry and Chemical Engineering, 30(2), 139–150.



Analytical Chemistry

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