6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 王德伟 *, 叶礼红, 杨祖伟 (, 519040) 摘要 : 目的 方法, C 18, 0.68% : (V:V=82:18), 1.0 ml/min, 340 nm 结果 D- 0.052 1.045 mg/ml, 97.5%, RSD%=1.2(n=9) 结论, 关键词 : ; ; ; Determination of N-Sulfo-glucosamine potassium salt by reversed-phase high performance liquid chromatography with online pre-column derivation WANG De-Wei *, YE Li-Hong, YANG Zu-Wei (By-Health Co., Ltd., Zhuhai 519040, China) ABSTRACT: Objective To establish a method for the determination of N-Sulfo-glucosamine potassium salt by reversed-phase high performance liquid chromatography (RP-HPLC) with online pre-column derivation in health food. Methods Online pre-column derivation were reacted by using phthalic dicarboxaldehyde (OPA) derived reagents and amino of N-Sulfo-glucosamine potassium salt. The products were separated by C 18 column with 0.68% acetic acid buffer: methanol (V:V=82:18) as mobile phase, the flow rate of 1.0 ml/min, and detection wavelength of 340 nm. Results D-glucosamine hydrochloride concentration had a good linearity in the range of 0.052~1.045 mg/ml. The average recovery rate was 97.5%, RSD%=1.2 (n=9). Conclusion The method is high, simple and accurate, and can be used to determine the content of N-Sulfo-glucosamine potassium salt in pharmaceutics and health food. KEY WORDS: N-Sulfo-glucosamine potassium salt; online pre-column derivatization; phthalic dicarboxaldehyde derived reagents; reversed-phase high performance liquid chromatography 1 引言 D-,,,, ;, * 通讯作者 :,, E-mail: 790007532@qq.com *Corresponding author: WANG De-Wei, Licensed Pharmacist, By-Health Co., Ltd., Zhuhai 519040, China. E-mail: 790007532@qq.com
8, : 3071, [1,2],,, [3-7] [8] [9] [10] [11-15],, reversed-phase high performance liquid chromatography (RP-HPLC), OPA,, 2 材料与方法 2.1 仪器与试剂 Agilent 1260 ( ); KQ-500E ( ); XW-80A ( ); XP-205 (Mettler Toledo ) OPA ( :, : BCBF1444); D- ( ); (, ); (, ); ( ) 2.2 色谱条件 : ZORBAX SB-C 18 5 μm, 4.6 mm 250 mm (Agilent); : 0.68% : =82:18 (V:V); : 1.0 ml/min; : 35 ; : 2 μl; : ; : 340 nm; : : OPA ( )2 μl; : 2 μl; : 6 ; : 1 min; : 2.3 溶液的制备 2.3.1 标准储备液 D-,, 1.0 mg/ml, 4 2.3.2 标准工作液 1.0 2.0 3.0 4.0 5.0 ml 10 ml,, 2.3.3 样品处理 0.2 g,, 50 ml, 3 min,,,, 2.4 线性试验 2.3.2,, (X), a β (Y),, 2.5 精密度试验 6, 2.3.3,, RSD(%) 2.6 加标回收率试验 0.1 g,, 3, 3, 3 D- 8 10 12 mg, 50 ml, 3 min,,,, 3 结果与讨论 3.1 GB/T 20365-2006 硫酸软骨素和盐酸氨基葡萄糖含量的测定液相色谱法 分析 GB/T 20365-2006 192 nm,,,,, 340 nm,,, 3.2 标准曲线验证结果 D- 1 0.999, 0.052 1.045 mg/ml
3072 6 表 1 标准曲线方程的结果 Table 1 The results of standard curve equation (r) D- Y=1868.9X+29.939 0.9999 3.3 精密度实验 D- 2, 6 (RSD) 1.0%, 表 2 精密度的实验 (n=6) Table 2 The precision of the experiment (n=6) (%) (%) RSD(%) 16.86 D- 16.59 16.78 16.67 1.0 16.79 16.44 16.58 3.4 回收率实验 D- 3 3, 3 D- : 97.5%, (RSD) 2.0%,, D- D- D- D- 3.5 实际样品的测定 2.3.3, D-, a β, 1,, 4 结论 RP-HPLC, OPA, GB/T 20365-2006 192 nm,,,,,,,, Table 3 表 3 加标回收率实验结果 (n=9) Standard addition recovery of experimental results (n=9) (mg) (mg) (%) (%) RSD(%) 8.032 7.813 97.07 8.025 7.796 97.15 8.003 7.693 96.13 10.112 9.905 97.95 D- 10.026 9.815 97.90 97.5 1.2 10.005 9.613 96.08 11.956 11.643 97.38 12.025 11.993 99.73 12.006 11.801 98.29
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