High Performance Liquid Chromatographic Method for the Analysis of Pyridoxine Hydrochloride in Liquid Dosage Form



  • Mohammed A. Salih Chemistry Department,College of Science, University of Sulaimani, Sulaimani. Iraq
  • Dlivan F. Aziz College of Pharmacy, University of Sulaimani, Sulaimani. Iraq
  • Salar I. Ali Nursing Department, Technical College of Health, Sulaimani Polytechnic University, Sulaimani Iraq


Pyridoxine hydrochloride, Reverse-phase HPLC, Stability, Verification


Vitamin B6 (pyridoxine hydrochloride) is available in the market as ampoule, but there is no simple method for its analysis. The aim of this study was to develop a relatively basic, very simple, fast, sensitive, accurate and less expensive HPLC method for the assessment of pyridoxine hydrochloride from vitamin B6 ampoule. The wavelength of absorbance of pyridoxine hydrochloride was measured at 280 nm. Mobile phase was prepared by dissolving 140 mg of sodium 1-hexanesulfonate per 100 mL of a mixture of methanol, glacial acetic acid and water (41:1:58). The mobile phase was transferred at a flow rate of 1.0 mL/min and the stationary phase used was a Waters Zorbax SB C18 column (5 μm pore size 250 * 4.6 mm). Injection volume was 20 μL/loop. The linearity was achieved with a linear regression (R2 ) = 0.9994 over the concentration range of 0.25-1 mg/mL. Separation was performed within 4.4min ± 0.10, no interference with any reported ingredients. Calculated percentage accuracy (recovery) was between 100.31 to 100.63%. The technique was discovered to be repeatable for inter- and intra-day precision level across the concentration of the sample with acceptable (RSD) NMT 2%. The technique have been verified and found suitable for determining pyridoxine hydrochloride in ampoule. This technique is useful in the quality control laboratory for routine use in the determination of pyridoxine hydrochloride in liquid dosage form. 


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How to Cite

Salih, M. A., Aziz, D. F., & Ali, S. I. (2020). High Performance Liquid Chromatographic Method for the Analysis of Pyridoxine Hydrochloride in Liquid Dosage Form: doi.org/10.26538/tjnpr/v4i12.5. Tropical Journal of Natural Product Research (TJNPR), 4(12), 1050–1055. Retrieved from https://www.tjnpr.org/index.php/home/article/view/836