4 3 Vol. 4 No. 3 2013 6 Journal of Food Safety and Quality Jun., 2013 B 6 何珊丽 *, 胡燕玲, 梁波 (, 528403) 摘要 : 目的 B 6 方法,, GB/T 5009.154-2003 B 6 结果, r 0.999 (S/N 3) 15 μg/kg 13 μg/kg 16 μg/kg 98.9%~99.5%, 0.88%~1.05% B 6, 结论 B 6, 关键词 : ( ); B 6 ; Detection of vitamin B 6 in beverage by high pressure liquid chromatography HE Shan-Li *, HU Yan-Ling, LIANG Bo (Zhongshan Supervision Testing Institute of Quality & Metrology, Zhongshan 528403, China) ABSTRACT: Objective To establish a rapid and effective method for the detection of vitamin B 6 in beverage by high pressure liquid chromatography (HPLC). Methods After dilution and filtration, samples was measured by high pressure liquid chromatography with fluorescence detector, and the data was compared with that obtained by the national standard microbial method. Results The method showed a good linearity with r 0.999. The LODs of pyridoxine, pyridoxal and pyridoxmine were 15, 13, 16 μg/kg, respectively. The recoveries were 98.9%~99.5% and the relative standard deviations were 0.88%~1.05%.The data of vitamin B 6 content in beverage detected by HPLC was consistent with that obtained by microbial method. But the detection time of HPLC was much shorter than that of the microbial method. Moreover, the operation of HPLC was simple, and the repeatability and accuracy of data were much higher as well. Conclusion HPLC is a sensitive method for the detection of vitamin B 6 in beverage, and it could be applied to the daily detection in laboratory. KEY WORDS: high pressure liquid chromatography (HPLC); vitamin B 6 ; beverage 1 引言, () B,,,, B, GB/T 5009.154-2003 B 6, 2~3 GB/T 5413.13-2010 * 通讯作者 :,, E-mail: sannyl@tom.com *Corresponding author: HE Shan-Li, Engineer, Zhongshan Supervision Testing Institute of Quality & Metrology, No.268, Zhongshan 3rd Road, Zhongshan 528403, China. E-mail: sannyl@tom.com
940 4 B 6, B 6, ( ) B 6,, B 6 ( ) 2 材料与方法 2.1 试剂与仪器 (); (0.5%) ( ); ( 99.99%)( Dr.Ehrenstorfer ); ( 99.99%)( Chem Service ); : 0.05 g( 0.0001 g), 50 ml,, 1 mg/ml : 1 ml, 50 ml,, 20 μg/ml ()(Agilent Technologies 1260, ); (BS-224S, Sartorius ): 0.0001 g 0.01 g (sartorius BS 2202S); (3-18K, Sigma ): 6000 r/min; 50 ml 2.2 色谱条件 C 18, 4.6 mm 150 mm ;, : 85:15( 2.0 g 2.5 ml, 1 L, ph 3.0±0.1);, 1.0 ml/min;, 30 ; : 293 nm, 395 nm;, 10 μl 2.3 样品处理方法 ( 2), 25 ml, 0.45 μm 3 结果与讨论 3.1 实验方法 0.0500g, 50mL( 1 mg/ml) 20mg/mL (2.2 ) 3.2 B 6 ( ) 1 Fig. 1 1 ( ) Chromatograms of mixed standard solution (pyridoxine, pyridoxal and pyridoxmine) 1 2 3 (1 pyridoxal 2 pyridoxine 3 pyridoxmine)
3, : B 6 941 3.2 方法的线性范围和检出限 0.0 1.0 2.0 3.0 5.0 ml,,, 0.00 0.20 0.40 0.60 1.00 μg/ml (A), (C, μg/ml), 0.015 ~2.00 μg/ml, : A =9.4296C 0.0001, A =13.599C 5E 05 A =10.769C 4E 05 (S/N=3) : 15 μg/kg 13 μg/kg 16 μg/kg r 0.999 3.3 方法回收率与精密度 10 g, 20 μg/ml 1 ml, 2.3, 5, 1 3 98.9%~ 99.5%, 0.88%~1.05%,, 3.4 样品的测定 2.3,, 2, 2n=6 X=6.40 mg/kg 3.5 方法比较 GB/T 5009.154-2003 B 6 B 6, 3, 4 表 1 回收率实验结果 (n=5) Table 1 Recovery of experiment results (n=5) B 6 (μg) (%) (%) 20 99.5 0.92 20 99.2 1.05 20 98.9 0.88 4 pyridoxine 2 Fig. 2 HPLC chromatogram of sample 表 2 高压液相色谱法测定样品中维生素 B 6 的含量 (n=6) Table 2 HPLC determination of the content vitamin B 6 in samples(n=6) (g) B 6 (μg/ml) B 6 X(mg/kg) 18.28 4.6726 6.39 15.22 3.8963 6.40 13.57 3.4793 6.41 15.65 4.0064 6.40 14.89 3.8059 6.39 13.92 3.5691 6.41
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