Kurioka S, Ishioka N, Sato J, Nakamura J, Ohkubo T, Matsuda M
Division of Biochemistry, School of Medicine, Jikei University, Tokyo, Japan.
Biomed Chromatogr. 1993 May-Jun;7(3):162-5. doi: 10.1002/bmc.1130070313.
A high performance liquid chromatographic (HPLC) procedure for measuring pyridoxal-5'-phosphate (PLP) and certain forms of B6 vitamers in plasma is presented here. This HPLC procedure consisted of a single graphitic carbon column with a fluorescence detector employing an isocratic eluent (15% acetonitrile: 1% perchloric acid: 0.05% sodium bisulfite). The graphitic carbon column is useful in acidic eluent without deteriorization. The relatively low fluorescent intensity of PLP under acidic conditions is improved by its derivatization with bisulfite in the eluent during chromatographic separation. Using this procedure, the detection limit of PLP is 50 fmol, and an aliquot of 5-50 microL of human plasma is required giving satisfactorily precise results within 5 min. We applied this method to the determination of PLP and certain B6 vitamers in human plasma after oral supplementation of pyridoxine.
本文介绍了一种用于测定血浆中磷酸吡哆醛（PLP）和某些形式维生素B6的高效液相色谱（HPLC）方法。该HPLC方法由一个带有荧光检测器的单石墨碳柱组成，采用等度洗脱液（15%乙腈：1%高氯酸：0.05%亚硫酸氢钠）。石墨碳柱在酸性洗脱液中使用时不会变质。在色谱分离过程中，通过在洗脱液中用亚硫酸氢钠对PLP进行衍生化，可提高其在酸性条件下相对较低的荧光强度。使用该方法，PLP的检测限为50飞摩尔，只需5-50微升人血浆的等分试样，即可在5分钟内获得令人满意的精确结果。我们将该方法应用于口服补充吡哆醇后人血浆中PLP和某些维生素B6的测定。
