6 5 Vol. 6 No. 5 2015 5 Journal of Food Safety and Quality May., 2015 陈彩云 *, 蔡伟江, 陈宏壁, 林贤邦 (, 519040) 摘要 : 目的, 方法, 0.005 mol/l,, Diamonsil C 18, 192 nm 结果,, 0.008 mg/ml~0.04 mg/ml, 1.000, 98.5%~100.0%, 0.6% 结论,,, 关键词 : ; ; ; Determination of chondroitin sulfate in health food by high performance liquid chromatography CHEN Cai-Yun *, CAI Wei-Jiang, CHEN Hong-Bi, LIN Xian-Bang (By-Health Co., Ltd., Zhuhai 519040, China) ABSTRACT: Objective To investigate a quick and simple method for determination of chondroitin sulfate in health foods by high performance liquid chromatography (HPLC). Methods Samples were treated using 0.005 mol/l pentane solution of sodium-acetonitrile as mobile phase with isocratic elution, separated on diamonsil C 18 column, and detected at 192 nm wavelength. Results Chondroitin sulfate could be separated well by this method. It had a good linear relationship within 0.008~0.04 mg/ml (r=1.000), the average recoveries was 98.5%~100.0%, and the RSD was 0.6%. Conclusion The method can be used for determination of chondroitin sulfate in health foods, it is quick, simple and accurate, and has a good reproducibility and clear separation, which can be popularized in the laboratory. KEY WORDS: high performance liquid chromatography; health foods; chondroitin sulfate; determination 1 引言,,,,,,, [1] : [2,3], [4], [5] [6-10],,,,, * 通讯作者 :, E-mail: 504990659@qq.com *Corresponding author: CHEN Cai-Yun, By-Health Co., Ltd., Zhuhai 519040, China. E-mail: 504990659@qq.com
1914 6 ;,,,,,,,, ;,,,,, ;,,,,,,,,,,,,,,,, 2 材料与方法 2.1 仪器与试剂 ( 1260, ); (EQ-500, ); Diamonsil C 18 (250 mm 4.6 mm, 5.0μm, Dikma) ( : USP; : H1K241; : 100%); (, CNW); (, ); ; ( ) 2.2 仪器条件 : Diamonsil C 18 (250 mm 4.6 mm, 5.0μm, Dikma); : +0.005 mol/l =5:95(v:v), ; : 0.8 ml/min; : 10 μl; : 192 nm [11] ; : 5 min 2.3 测定方法 2.3.1 标准系列溶液的配制 105 10 mg, 50 ml, 5 ml, 10 min,,,, 1 2 3 4 5 ml 25 ml,,, 0.008 0.016 0.024 0.032 0.04 mg/ml, 0.45 μm C(mg/mL), (A), 2.3.2 供试品溶液配制 20,, 0.1 g, 50 ml, 5 ml, 10 min,,,, 0.45 μm, 3 结果与讨论 3.1 色谱行为, 0.005 mol/l, 5 min,, 1 2,,,, 3 4 3.2 线性实验 1, r 2 1.0000,, 0.008 mg/ml~0.04 mg/ml 3.3 定量限和检出限 (detection limit, DL) (quantitation limit, QL) (S/N) DL S/N=3, QL S/N=10
5, : 1915 Fig. 1 1 Chondroitin sulfate standard of chromatogram map Fig. 2 2 The chromatogram map of the sample Fig. 3 3 The chromatogram map of the blank reagent Fig. 4 4 The chromatogram map of the excipients
1916 6 表 1 线性实验结果 Table 1 Linear experimental results C(mg/mL) (A) STD1 0.0080 477.70709 STD2 0.016 959.16284 STD3 0.024 1444.64282 STD4 0.032 1929.9469 STD5 0.040 2404.77222 Y=60311X-4.2279 (r 2 ) 1.0000 3.3.1 定量限 0.2328 mg/ml 1~100 ml,, 2.328 μg/ml, 10, S/N=10 : QL 2.328 μg/ml*50 ml/0.1 g=1.164 mg/g 3.3.2 检出限 S/N=3, DL QL/10*3= 1.164/10*3 mg/g=0.3492 mg/g 3.4 稳定性实验 0 2 4 8 12 24 h, RSD=0.62%, 24 h 3.5 精密度实验 6, 2.3.2,, RSD(%) 6 6.01 g/100 g, (RSD) 0.1%,, 2 3.6 加标回收率实验 0.1 g 9, 50 ml, 3, 3, ( 0.816 mg/ml) 6.0 ml 7.5 ml 9.0 ml,, 10 min,, 0.45 μm,,, 3 Table 2 表 2 精密度实验结果 The precision of experimental results (g) (mg/ml) (g/100g) (g/100g) RSD(%) 1 0.10261 0.123455 6.015 2 0.10265 0.12326 6.003 3 0.10258 0.123371 6.013 4 0.10267 0.123376 6.008 5 0.10269 0.123400 6.008 6 0.10272 0.123424 6.007 6.01 0.10% Table 3 表 3 加标回收率实验结果 Standard addition recovery experiment results (g) (mg/ml) (mg) (mg) (%) (%) RSD(%) 1 0.10701 0.226226 4.879999 4.896 99.673 2 0.10598 0.224018 4.831502 4.896 98.682 3 0.10293 0.221664 4.897107 4.896 100.022 4 0.11042 0.25409 6.068258 6.120 99.154 5 0.10948 0.252117 6.026102 6.120 98.465 6 0.10953 0.253581 6.096297 6.120 99.612 7 0.10421 0.271596 7.316779 7.344 99.629 8 0.10425 0.271731 7.321125 7.344 99.688 9 0.10429 0.271628 7.313571 7.344 99.585 99.39 0.6
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