Microsoft Word 虞哲高-二校出1907

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1 6 5 Vol. 6 No Journal of Food Safety and Quality May., 2015 C 虞哲高 *, 林冰东 *, 邢月寒, 王晓燕 (,, ) 摘要 : 目的 C, C 方法 Waters Sep-Pak C 18, D101 (SPE) C,,, 510 nm 结果 0~48 µg/ml, , (RSD) 5%, 95.31%~ 98.38% C (Vc: =1:1, 2:1, 10:1), C,,, 结论, C 关键词 : C; ; ; D101 ; Determination of total flavonoids in rich ascorbic acid food YU Zhe-Gao *, LIN Bing-Dong *, XING Yue-Han, WANG Xiao-Yan (DSM Nutritional Products China Research Center, DSM China Co., Ltd., Shanghai , China) ABSTRACT: Objective To establish a quantitative analysis of total flavonoids with high content of ascorbic acid(vitamin C) in food and solve the interference of ascorbic acid on the measurement of total flavonoids by complexation reaction. Methods A solid phase extraction procedure was developed to degrease and remove pigments by Sep-Pak C 18, vitamin C was removed with D101 macro-porous resin, total flavonoids were determined by aluminum nitrate colorimetric using detection wavelength at 510 nm. Results The developed method was successfully used to determine total flavonoids in vitamin C rich fruits/plant extracts. It showed a good linearity in the range of 0~48 μg/ml with r= The recoveries were 95.31%~98.38% at 3 spiked levels, and the relative standard deviations (RSDs) were less than 5%. Three kinds of food (ascorbic acid:flavonoids=1:1, 2:1, 10:1) were analyzed, which showed that higher ascorbic acid content could affect the testing result. So, it is necessary to remove the ascorbic acid before coloration in rich ascorbic acid products. Conclusion The method is suitable for simple, environmental, and effective determination of total flavonoids in rich ascorbic acid foods. KEY WORDS: vitamin C; total flavonoids; solid phase extraction; D101 macro-porous resin; aluminum nitrate colorimetric * 通讯作者 :,, / Zhe-Gao.Yu@dsm.com,,, linda.lin@dsm.com *Corresponding author: YU Zhe-Gao, Master, Nutrition Analytics Executive, Nutritional Products China Research Center, DSM China Co., Ltd., Shanghai , China. Zhe-Gao.Yu@DSM.com LIN Bing-Dong, Scientist Analysis, Nutritional Products China Research Center, DSM China Co., Ltd., Shanghai , China. linda.lin@dsm.com

2 引言 2-,,,,, [1-3], ( ) [4] ( ) [5] [6] [7],,,,, [6],,,,,,,, 510 nm, C, C C,, [8] C, [5], C C, C, C 2 材料与方法 2.1 材料和仪器 ( 92.8%, ); ( 98%, ); ( 98% ); ( 98%, ); ( 98%, ); (, ); (, ); (, ); (, ); ( ); (, ); ( ); (Swisssur GmbH ); (Maruei ) - (HELIOS ZETA, Thermo Scientific ); (AT 261, Mettle Toledo ); (SPD 2010, Thermo Scientific ); (Multifuge X3R, Thermo Scientific ); (DK-S14, ); (SS680-B, ); (LSC, CHRIST ); -80 (FORMA700 series plus, Thermo Scientific ); HPLC 1260 ( Agilent ); Sep-Pak-C 18 (500 mg/3 ml, Waters ); D101 (500 mg/6 ml, ); Oasis HLB cartridge(500 mg/6 ml, Waters, ); Discovery DPA-6s (500 mg/6 ml, SUPELCO, ); 2.2 分析方法标准曲线的制备 10 mg,, 50 ml, 40 ml, 10 min, 50 ml ml, 50 ml,, 20 ml (5 g/100 ml)2 ml,, 6 min; (10 g/100 ml)2 ml, 6 min; 20 ml 1 mol/l,,, 15 min,

3 5, : C nm,, 样品的前处理 (1) 2 g( 350 mg ),,,,, 30 ml 70%,, 80 2 h 50 ml, 70%,, 70%, 4000 r/min 10 min, A 10 ml A Sep-Pak-C 18, 5 ml, 5 ml, B 2.0 ml B 50 ml 70% 2.0 ml B,, 60, 2 ml, D101 Sep-Pak-C ml,, 20 ml 50 ml, (5 g/100 ml) 2 ml, (2) 20 ml, h,, 分析结果计算, C, : C 1250 X m 10 : X ( ), g/100 g C, µg/ml m, g 1250, ml 高效液相色谱 (HPLC) 条件 : Hydrosphere C 18 (150 mm 4.6 mm I.D.), S-5 µm, YMC ); : 25 ; : 254 nm; : 1.0 ml/min; : 5 µl; : 0.025%TFA A, 0.025%TFA B, : 0 10 min, A 70% 20%; min, A 20%; min, A 20% 0%; min, A0%; min, A 0% 70% 3 结果和讨论,, 510 nm,, C, C D101,, 样品前处理的优化黄酮类物质在不同固相萃取小柱吸附和洗脱能力, D101 [9,10], Oasis HLB [11] [5] 5 1 mg, 5 10 ml, 5mL 5mL,, 2.2.4, 1 5 D101, 90% 高含量维生素 C 对总黄酮测定的影响及其消除 ( : C=1:7.5, 2.40 g/100 g), 70%, Waters Sep-Pak C 18, g/100 g, 4.7% D101, Waters Sep-Pak C 18, g/100 g, 3.5%, D101,

4 Table 1 表 1 3 种不同固相萃取小柱回收率比较 (HPLC) Comparison of 3 different solid phase extraction recovery % D101 Oasis HLB (Discovery DPA -6s) 90.05% 84.56% 32.76% 96.10% 99.46% 35.01% 92.60% 88.06% 28.83% % 44.09% 23.64% % 46.00% 52.67% D101 D101 HPLC,, D101, C ( 1) 2, Y=0.0151X , r=0.9998, 0 48 µg/ml, 2 Fig. 2 The standard curve of rutin 1 D101 Fig. 1 Chromatogram of lavonoids with ascorbic acid solution before and after D101 macroporous resin solid phase extraction, D101,,, C D101 C, D101 C 3.2 线性范围 510 nm, (Y), (X),, 3.3 精密度实验 2.2, ( : C=1:7.5, 2.40 g/100 g), 2 2,, 6 5%, C 3.4 加标回收率实验 ( : C=1:7.5), 2.4 g/100 g,, 3,

5 5, : C 1911 表 2 精密度实验结果 (n=6) Table 2 Results of precision test (n=6) (g/100 g) (g/100g) RSD(%) % 表 3 Table 3 加标回收率实验结果 (n=3) Results of recovery test (n=3) : (%) (%) 2: : : Table 4 表 4 3 种食品总黄酮测定结果 ( 分光光度法 ) Results of total flavonoids content of 3 kinds of foods A B C (v/v) A (g/100 g) B (g/100 g) 1: % 2: % 10: % : A D101, 2.2 B D101, Waters Sep-Pak C 18 SPE 3 3 : =2:1, : =1:1, : =1: % 98.38%, 3.03% 5.16%, 实际样品检测结果 3,, 4, C, C,, C, C, C, 3, 3, : C=1:7.5, 2.4 g/100 g 2.21 g/100 g, 2.33 g/100 g 2.26 g/100 g 4 结论 C,, Waters Sep-Pak C 18 D101, C C,,,,, 90%, C, 参考文献 [1] Megan EW, Lisa J. Mauer. Development of an integrated approach for the stability testing of flavonoids and ascorbic acid in powders [J]. Food Chem, 2011, 129: [2],,. [J].

1912 6, 2007, 17(6): 1037. Xiao XL, Luo RC, Zhang N. Determination of total flavonoids in health food [J]. Chin J Health Lab Technol, 2007,17(6): 1037. [3],,. [J]., 2002, 12(4): 401 402.

Liu JX, Li Y, Lu XN, et al. Study on the extraction technology and determination of the flavonoid in health food [J]. Food Res Devel, 2008, 29(9): 116 118. [5]. [M]. :, 2002. Wang GY.

6 1912 6, 2007, 17(6): Xiao XL, Luo RC, Zhang N. Determination of total flavonoids in health food [J]. Chin J Health Lab Technol, 2007,17(6): [3],,. [J]., 2002, 12(4): Yu C, Yu S, Shen XH. Study on determination method of general flavone in health foods [J]. Chin J Health Lab Technol, 2002, 12(4): [4],,,. [J]., 2008, 29(9): Liu JX, Li Y, Lu XN, et al. Study on the extraction technology and determination of the flavonoid in health food [J]. Food Res Devel, 2008, 29(9): [5]. [M]. :, Wang GY. Determination of efficacy in health food [M]. Beijing: China Light Industry Press, [6],,,. [J]. -, 2007, 43: , 467. Peng YF, Ma YH, Guo YD, et al. Rapid determination of flavonoid in Saururus chinensis (Lour) baill by high performance liquid chromatography [J]. Phys Test Chem Anal, (Part Chem Anal), 2007, 43: , 467. [7],,,. [J]., 2007, 39(12): Chi YM, Yu S, Guo R, et al. Determination of flavonoids from radix ranunculi ternati with difference spectrophotometry [J]. Jiangsu J Tradit Chin Med, 2007, 39(12): [8],,,. C [J]., 2004, 10(6): Cui NJ, Cui NQ, Fu Q, et al. Bidirectional effect of vitamin C on pro-oxidation/antioxidation [J]. Chin J Surg Integrated Tradit West Med, 2004, 10(6): [9],,. [J]., 2010, 21(1): 8 9. Pei YP, Li WL, Zhang HQ. The property of the macroporous resin for isolation of total flavones from Gynura bicolor DC [J]. Lishizhen Med Mater Med Res, 2010, 21(1): 8 9. [10],,,. D-101 [J]., 2010, 31(8): Hu ZJ, Hao LJ, Wang NX, et al. Purification of crude total flavonoids extracted from citrus peel using D-101 macroporous adsorption resin [J]. Food Sci, 2010, 31(8): [11],,,. 5 [J]., 2013, 34(6): Liang C, LuH X, Liu HC, et al. Establishment of determination method for flavonoids in honey and distribution of flavonoids in 5 kinds of characteristic honey from Yunan [J]. Food Sci, 2013, 34(6): 作者简介 ( 责任编辑 : 李振飞 ) 虞哲高, 硕士, 主要研究方向为食品 / 饲料检测 Zhe-Gao.Yu@DSM.com 林冰东, 博士, 分析研究员, 主要研究方向为天然产物相关食品及保健品的分析和检测 linda.lin@dsm.com

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min 25 4 2013 8 监测技术 5 1 2 1 ( 1. 江苏省疾病预防控制中心, 江苏南京 210009; 2. 南京医科大学第二附属医院, 江苏南京 210029) 5 Phenomenex kinetex C 18 2. 6 μm 4. 6 mm 150 mm 10 mmol /L ph 2. 5 + 60 40 DAD 280 nm 5 20. 0 mg /L ~ 100 mg /L 0.