高效液相色谱法测定肾病综合征患者血清泼尼松及其临床意义
HPLC determination and clinical significance of serum prednisone in patients with nephrotic syndrome.
作者信息
Chen Chun-Mei, Xia Yun-Cheng, Zhang Xu-Guang, Peng Can-Hui, Liu Fu-You, Peng You-Ming, Sun Lin
机构信息
Institute of Kidney Disease, The Second Xiangya Hospital, Central South University Changsha, Hunan, China ; Clinical Laboratory, The Second Xiangya Hospital, Central South University Changsha, Hunan, China.
Institute of Kidney Disease, The Second Xiangya Hospital, Central South University Changsha, Hunan, China.
出版信息
Int J Clin Exp Med. 2014 Dec 15;7(12):5517-22. eCollection 2014.
AIM
A rapid protocol is necessary to determine the serum concentrations of prednisone.
METHODS
The HP1100 high-performance liquid chromatographic (HPLC) system was employed. The HP Lichrosphere C8 column (250 mm × 4 mm, i.d., 5 μm particle size) was used. The mobile phase was methanol, tetrahydrofuran and water in the ratio 25:25:50. The flow rate was 1.0 ml/min. The sample was monitored by UV absorbance at 240 nm. Acetanilide was used as the internal standard, and methanol was added into the serum for depositing the protein.
RESULTS
The chromatography was effective and was not interfered with by the serum components. Good linearity was observed, within the range of 10-500 μg/L for prednisone, and the detection limit was 5 μg/L. The serum concentrations of prednisone between the nephrotic syndrome (NS) group and the control group were significantly different (P < 0.05), while there was no significant difference between the females and males of the NS group (P > 0.05). The serum ncentration of prednisone in the steroid-resistant group was lower than that in the steroid-sensitive group (P < 0.05).
CONCLUSIONS
HPLC is a practical and reliable method to determine the serum concentration of prednisone with high accuracy, precision, linearity and repeatability.
目的
需要一种快速方案来测定泼尼松的血清浓度。
方法
采用惠普1100高效液相色谱（HPLC）系统。使用惠普Lichrosphere C8柱（250 mm×4 mm，内径，5μm粒径）。流动相为甲醇、四氢呋喃和水，比例为25:25:50。流速为1.0 ml/min。样品通过在240 nm处的紫外吸光度进行监测。使用乙酰苯胺作为内标，并向血清中加入甲醇以沉淀蛋白质。
结果
该色谱有效，不受血清成分干扰。观察到良好的线性关系，泼尼松浓度在10 - 500μg/L范围内，检测限为5μg/L。肾病综合征（NS）组与对照组的泼尼松血清浓度有显著差异（P < 0.05），而NS组的女性和男性之间无显著差异（P > 0.05）。激素抵抗组的泼尼松血清浓度低于激素敏感组（P < 0.05）。
结论
HPLC是一种实用且可靠的方法，可高精度、精密度、线性和重复性地测定泼尼松的血清浓度。
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