8 5 Vol. 8 No. 5 2017 5 Journal of Food Safety and Quality May., 2017 B 1 B 2 B 6 陈彩云 *, 魏鲜娥, 蔡伟江, 黎小兰, 张喜金 (, 519040) 摘要 : 目的 B 1 B 2 B 6 方法 0.2%, Alltima-C 18, -5 mmol/l, (diode array detector, DAD) 200 nm 280 nm 结果 B 1 B 2 B 6,, 93.9%~103.2%, (relative standard deviation, RSD) 0.4%~1.8% 结论, B 1 B 2 B 6 关键词 : ; ; B 1 ; B 2 ; B 6 ; Determination of pantothenate, nicotinamide, vitamin B 1, vitamin B 2, and vitamin B 6 in healthy food by high performance liquid chromatography CHEN Cai-Yun *, WEI Xian-E, CAI Wei-Jiang, LI Xiao-Lan, ZHANG Xi-Jin (By-Health Co., Ltd., Zhuhai 519040, China) ABSTRACT: Objective To establish a method for the simultaneous determination of pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 in healthy food by high performance liquid chromatography (HPLC). Methods Samples were ultrasonically extracted with 0.2% phosphoric acid solution, separated by alltima-c 18 column with acetonitrile-5 mmol/l sodium heptanesulfonate solution as the mobile phase for gradient elution, and then detected by diode array detector (DAD) simultaneous at the wavelengths of 200 nm and 280 nm. Results Pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 could be completely separated, and they also had good linear relationships in their linear ranges. The average recoveries were 93.9%~103.2%. The relative standard deviations were 0.4%~1.8%. Conclusion This method is rapid, simple, accurate and repeatable, which can be used for the simultaneous determination of pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 in healthy food. KEY WORDS: pantothenate; nicotinamide; vitamin B 1 ; vitamin B 2 ; vitamin B 6 ; high performance liquid chromatography * 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
5, : B 1 B 2 B 6 1867 1 引言,,,,,,, A B 2, B 1 C,,,, (high performance liquid chromatography, HPLC) [1-10], 1, B 6 B 2 B 1 5, 5,,,, 2 材料与方法 2.1 仪器与材料 Agilent 1260 (, (diode array detector, DAD)); XP 205 ( - ); KQ-500VDE ( ) B, 20150601D 20150602D 20150603D, (, ) (, ) (, ); ( : 99.9%, Supelco, : LC10011V); ( :99.9%, Supelco, :LC01839V); B 1 ( : 99.9%, Supelco, : LB84083V); B 2 ( : 98.7%, Dr.Ehrenstorfer GmbH, : D0144); B 6 ( : 99.9%, Supelco, : LB83988V) 2.2 试验方法 2.2.1 色谱条件 : Alltima C 18 (4.6 mm 150 mm, 5 μm); : 1.0 ml/min; : 40 ; : 10 μl; : 200 nm 280 nm; : (diode array detector, DAD); : A 5 mmol/l, B, 1 表 1 梯度洗脱程序 Table 1 Gradient elution program (min) A (%) B (%) 0.00 95 5 15.00 77 23 15.01 95 5 20.00 95 5 2.2.2 标准溶液的制备 (1) 12 mg 35 mg 5 mg B 1 5 mg B 2 5 mg B 6 ( 0.01 mg), 100 ml, 1 ml, ( ),, 0.45 μm, 4 (2), 0.45 μm, 10 μl, c(mg/ml), A, 2.2.3 样品前处理 20,,, 1.0 g 50 ml, 0.2%, 40 ( 500 W, 50 khz) ( 25 min),, 0.2%,,, 0.45 μm, 3 结果与分析 3.1 溶剂的选择 5, B 1 B 6 4, B 2, B 2,,, B 2, 0.2%,,,, 0.2% 3.2 流动相的选择 5,,,, C 18,, -, -, -, -, - -,,
1868 8,,,,,, 5 1.5,,,, 0.005 mol/l, 5,,,,,, B 1 B 2 B 6 5,, -,, ( 1) 3.3 波长的选择,,, 4 5, 200 [11] 260 280 [12] 444 [13] B 1 B 2 B 6 ( 2),, 200 nm,, ; 260 280 444 nm 200 280 nm, B 1 B 2 B 6, 260 nm, B 6, 444 nm B 1 B 6 200 280 nm, 5, 200 nm 280 nm B 1 B 2 B 6 3.4 方法学考察 3.4.1 方法的线性,, B 1 B 2 B 6 0.004~0.60 mg/ml, R 0.9995, 2 3.4.2 检出限与定量限, 3 (limit of detection, LOD), 10 (limit of quantification, LOQ), ( 3) 3.4.3 精密度试验, 6,, B 6 B 2 B 1, (relative standard deviations, RSD) 0.1% 1 B 1 B 2 B 6 Fig. 1 Gradient elution chromatograms of calcium pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6
5, : B 1 B 2 B 6 1869 Fig. 2 2 B 1 B 2 B 6 Chromatograms of calcium pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 at different wavelengths 表 2 泛酸钙 烟酰胺 维生素 B 1 维生素 B 2 和维生素 B 6 的线性关系 Table 2 Linear relationships of calcium pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 R (mg/ml) Y=11941X+11.25 1.0000 0.011918~0.11918 Y=1715X+9.385 0.9999 0.058052~0.58052 B 6 Y=16716X+2.389 1.0000 0.0046154~0.046154 B 2 Y=29822X+2.395 1.0000 0.0055963~0.055963 B 1 Y= 7896X+0.046 1.0000 0.0050649~0.050649
1870 8 表 3 泛酸钙 烟酰胺 维生素 B 1 维生素 B 2 和维生素 B 6 的检出限与定量限 (n=6) Table 3 Limits of detection and limits of quantification of calcium pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 (n=6) (μg/ml) (μg/ml) 0.03575 0.1192 0.8708 2.9026 B 6 0.06923 0.2308 B 2 0.08394 0.2798 B 1 0.07597 0.2532 B 2 B 6 5, 表 5 加标回收率和相对标准偏差 (n=3) Table 5 The recoveries and RSDs (n=3) (mg) (%) RSD(%) 1.21 102.2 0.5 1.45 103.2 0.4 1.81 102.2 1.1 2.81 95.5 1.0 3.37 98.4 0.4 3.4.4 重复性试验 B, 6, B 6 B 2 B 1, RSD 4 表 4 泛酸钙 烟酰胺 维生素 B 1 维生素 B 2 和维生素 B 6 5 种成分的重复性 (n=6) Table4 Repeatabilities of calcium pantothenate, nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 (n=6) (mg/100 g) RSD(%) 666.49~670.30 0.4 1660.59~1690.41 0.6 B 6 266.07~269.75 0.3 B 6 B 2 B 1 4.21 98.6 0.9 0.405 94.5 0.7 0.486 96.4 0.7 0.608 96.4 1.0 0.408 98.6 1.5 0.490 96.7 0.7 0.613 97.3 1.4 0.402 93.9 1.2 0.482 95.5 0.6 0.602 97.7 1.8 B 2 247.00~249.82 0.5 B 1 276.06~279.45 1.2 3.4.5 方法回收率 9, 3, 3, 50 ml, 0.2%, 1.21 1.45 1.81 mg; 2.81 3.37 4.21 mg B 1 0.402 0.482 0.602 mg B 2 0.408 0.490 0.613 mg B 6 0.405 0.486 0.608 mg,, 5 93.9%~103.2%, RSD 0.4%~1.8%, B 1 B 2 B 6 4 结论 HPLC, B 1 B 2 B 6,, B 1 参考文献 [1]. HPLC (15) B 1 B 2 B 6 [J]., 2012, 15(5): 6 10. Wang WJ. Determination of nicotinamide, vitamin B 1, vitamin B 2 and vitamin B 6 in Duoweiyuansu capsules (15) by HPLC [J]. China Pharm, 2012, 15(5): 6 10 [2]. B1 B2 [J]., 2016, 16(8): 1070 1072. Zeng YM. Content determination of vitamin B 1, vitamin B 2 in compound hepar extrectum tables by HPLC [J]. Eval Anal Drug-Use Hosp China, 2016, 16(8): 1070 1072. [3]. HPLC 5 B [J]., 2016, (1): 21 23, 38. Liang L. Simultaneous determination of 5 kinds of vitamin B in vitamin premix feed by HPLC [J]. China feed, 2016, (1): 21 23, 38. [4]. HPLC B B 1 B 2 B 6 [J]., 2012, 26(8): 891 894. Han XM. Determination of vitamin B 1, vitamin B 2 and vitamin B 6 in compound vitamin B tablets by HPLC [J]. Chin Pharm Aff, 2012, 26(8): 891 894. [5]. HPLC B 1 B 2 [J]., 2015, 26(18): 2549 2551.
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