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Validation of HPLC-UV Method for Analysis of 4-Pyridoxic Acid in Human Urine

Journal Name:

  • International Journal of Medical Research & Health Sciences

Publication Year:

  • 2017

Key Words:

Author NameUniversity of Author
Abstract (2. Language): 
We present a simple and validated HPLC method for measuring Vitamin B6 urinary metabolite 4-pyridoxic acid (4-PA). Urine samples were treated with 6% perchloric acid. HPLC separation was achieved using Waters column (Symmetry® C18 250 mm × 4.6 mm i.d (5 μM). The mobile phase was a mixture of methanol and 35 mM sodium phosphate buffer containing 2.5 mM sodium heptane polysulfonate at pH=3.5 with 85% O-phosphoric acid. Retention times were 8.0 min for 4-pyridoxic acid. Detection (UV at 302 nm) of analyte was linear in a range from 0.0125 to 0.8 μM. This method is stable, reproducible and has practical advantages such as ease and low cost.
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  • English

REFERENCES

References: 

[1] Kimura, Mieko, Kazunori Kanehira, and Katsuhiko Yokoi. “Highly sensitive and simple liquid chromatographic
Shawaqfeh, et al. Int J Med Res Health Sci 2017, 6(12): 100-104
104
determination in plasma of B6 vitamers, especially pyridoxal 5′-phosphate.” Journal of Chromatography A Vol.
722, No. 1-2, 1996, pp. 295-301.
[2] Dolina, Svetlana, et al. “Epilepsy as a pyridoxine-dependent condition: Quantified urinary biomarkers for status
evaluation and monitoring antiepileptic treatment.” Medical Hypotheses Vol. 79, No. 2, 2012, pp. 157-64.
[3] Bates, Christopher J., et al. “A simple, sensitive and reproducible assay for pyridoxal 5′-phosphate and 4-pyridoxic
acid in human plasma.” Clinica Chimica Acta Vol. 280, No. 1, 1999, pp. 101-11.
[4] Bisp, Marianne R., et al. “Determination of vitamin B6 vitamers and pyridoxic acid in plasma: development and
evaluation of a high-performance liquid chromatographic assay.” Analytical Biochemistry Vol. 305, No. 1, 2002,
pp. 82-89.
[5] Rybak, Michael E., and Christine M. Pfeiffer. “Clinical analysis of vitamin B 6: determination of pyridoxal
5′-phosphate and 4-pyridoxic acid in human serum by reversed-phase high-performance liquid chromatography
with chlorite post-column derivatization.” Analytical Biochemistry Vol. 333, No. 2, 2004, pp. 336-44.
[6] Marszałł, Marcin L., et al. “Application of the high-performance liquid chromatography method with coulometric
detection for determination of vitamin B 6 in human plasma and serum.” Journal of Chromatography B Vol. 877,
No. 27, 2009, pp. 3151-58.
[7] U.S. Department of Health and Human Services Food and Drug Administration, Center for Drug Evaluation
and Research (CDER), Center for Veterinary Medicine (CVM). Guidance for Industry, Bioanalytical Method
Validation, May 2001.

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