Chun-Mei Chen1, Yun-Cheng Xia2, Xu-Guang Zhang3, Can-Hui Peng4, Fu-You Liu2, You-Ming Peng2, Lin Sun2. 1. 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. 2. Institute of Kidney Disease, The Second Xiangya Hospital, Central South University Changsha, Hunan, China. 3. Clinical Laboratory, Affiliated Hospital, Weifang Medical College Weifang, Shandong, China. 4. Clinical Laboratory, The Third Xiangya Hospital, Central South University Changsha, Hunan, China.
Abstract
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.
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.