1Department of Pharmacy Practice, King Saud Bin Abdulaziz University for Health Sciences, Riyadh, KSA
2School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania, USA
3College of Pharmacy, Nova Southeastern University, Florida, USA
*Corresponding e-mail: shawaqfehm@ksau-hs.edu.sa
Online published on 19 November, 2018.
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.
Vitamin B6, Validation, 4-pyridoxic acid, HPLC, Urine
