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KMID : 1134820170460101186
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Journal of the Korean Society of Food Science and Nutrition
2017 Volume.46 No. 10 p.1186 ~ p.1194
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Quantitative Analysis of Vitamin B5 and B6 Using High Performance Liquid Chromatography
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Kim Gi-Ppeum
Hwang Young-Sun
Choung Myoung-Gun
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Abstract
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Recently, many people have demanded reliable nutritional data even for minor-components. On the other hand, an analytical method for the analyses of vitamin B5 and B6 is lacking. Therefore, this study attempted to validate with accuracy and precision the analysis of vitamin B5 and B6 using a high-performance liquid chromatography (HPLC) method. The vitamin B5 and B6 contents were analyzed using an Agilent 1260 series HPLC system. YMC-Pack ODS-AM (250×4.6 mm I.D.) and YMC-Pack Pro RS C18 (250×4.6 mm I.D.) columns were used for the analyses of vitamin B5 and B6, respectively. In the case of vitamin B5, the flow rate was set to 1.0 mL/min by isocratic elution using the 50 mM KH2PO4 solution (pH 3.5)/acetonitrile (ACN) (95:5, v/v) with monitoring at 200 nm using HPLC/DAD, whereas the flow rate for vitamin B6 was set to 1.0 mL/min of flow rate by isocratic elution using a 20 mM CH3CO2Na solution (pH 3.6)/ACN (97:3, v/v) with monitoring by excitation at 290 nm and emission at 396 nm using HPLC/FLD. The column temperature was set to 30°C. The injection volume was 20 μL for each experiment. The specificity of the accuracy and precision for vitamin B5 and B6 were also validated by HPLC. The results showed high linearity in the calibration curve for vitamin B5 (R2=0.9998**), the limit of detection (LOD) and limit of quantitation (LOQ) were 0.4 mg/L and 1.3 mg/L, respectively, In contrast, for the calibration curve of vitamin B6, which showed high linearity (R2=0.9999**), the LOD and LOQ were 0.006 mg/L and 0.02 mg/L, respectively.
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KEYWORD
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vitamin B5, vitamin B6, HPLC/DAD, HPLC/FLD
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