고속액체크로마토그래피를 이용한 비타민 B5 및 B6의 정량 분석

• Published in: Han'guk Sikp'um Yŏngyang Kwahakhoe chi Vol. 46; no. 10; pp. 1186 - 1194
• Main Authors: 김기쁨(Gi-Ppeum Kim), 황영선(Young-Sun Hwang), 정명근(Myoung-Gun Choung)
• Format: Journal Article
• Language: Korean
• Published: 한국식품영양과학회 01.10.2017
• Subjects: 식품과학; HPLC/FLD; 5< 6< HPLC/DAD; vitamin B< SUB&gt
• ISSN: 1226-3311; 2288-5978

식품 함유 비타민 B5 및 B6의 최적 HPLC 분석 조건을 검토한 결과 비타민 B5의 경우 YMC-Pack ODS-AM(250×4.6 mm I.D.) 칼럼을 이용하고, A용매로 50 mM KH2PO4(pH 3.5)을, B용매는 아세토니트릴을 이동상 용매로 사용하는 A용매 95% 등용매용리 조건에서 200 nm의 파장으로 분석하는 HPLC/DAD법을 최적조건으로 확립하였다. 한편 비타민 B6의 최적분석조건은 여기파장(excitation) 290 nm, 방출파장(emission) 396 nm로 분석하는 HPLC/FLD법으로써, 칼럼은 YMC-Pack Pro RS C18(250×4.6 mm I.D.), 이동상 용매는 A용매 20 mM CH3CO2Na(pH 3.6), B용매 아세토니트릴을 A용매 97% 등용매용리 조건으로 사용하였다. 비타민 B5 및 B6의 표준검량선은 R2값이 각각 0.9998 및 0.9999로 고도의 직선성을 나타내었고, 검출한계 및 정량한계는 비타민 B5의 경우 각각 0.4 mg/L 및 1.3 mg/L, 비타민 B6의 경우 각각 0.006 mg/L 및 0.02 mg/L로 산출되었다. 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. KCI Citation Count: 5