Determination of azidothymidine and its degradation product thymine in pharmaceutical dosage forms by HPLC and HPTLC densitometry


HPTLC densitometry and HPLC are considered for the determination of azidothymidine and its degradation product thymine in pharmaceutical dosage forms. In HPTLC the substances were separated on silica gel with fluorescence indicator in methanol-chloroform (10∶90) and methanol-chloroform (15∶85) systems. Absorbance measurement (detection of reflectance) of the separated substances was carried outin situ at 268 nm using four-level calibration (external standard, linear regression function) in the concentration range of 25–100 ng thymine/spot and using single-level calibration (external standard) at the concentration of 100 ng azidothymidine/spot. HPLC was carried out using RP-18 stationary phase and methanol+aqueous 0.03 mol/l KH2PO4 (18+82, v/v) as the mobile phase. The temperature was 50°C and the detection wavelength 266 nm. The detection limit of thymidine was 0.05%. The concentration range for azidothymidine was 0.5–1.5 mg/ml and for thymine 1–40 μg/ml (for an injection volume of 10 μl). The results were evaluated by linear regression analysis.

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Tománková, H., Šabartová, J. Determination of azidothymidine and its degradation product thymine in pharmaceutical dosage forms by HPLC and HPTLC densitometry. Chromatographia 29, 44–50 (1990).

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Key Words

  • Column liquid chromatography
  • Thin layer chromatography
  • Azidothymidine
  • Thymine