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1 6 4 Vol. 6 No Journal of Food Safety and Quality Apr., 2015 C 马丽 1*, 白淑荣 2, 韩占友 1, 王佳 1, 王蕾 1 (1., ; 2., ) 摘要 : 目的 C 方法 Oasis HLB,,, C,, C 结果 Y=0.0118X , R 2 =0.9989, : 760 nm, : (1:1), 7.5% Na 2 CO 3, 20~30, 60 min(re) (WE),, (HWE), WE C 结论,, C 关键词 : ; ; C; Study on optimized determination of total polyphenols and vitamin C MA Li 1*, BAI Shu-Rong 2, HAN Zhan-You 1, WANG Jia 1, WANG Lei 1 (1. Food and Drug Control Institution of Hulun Buir, Hulun Buir , China; 2. Hulun Buir Peoples Hospital, Hulun Buir , China) ABSTRACT: Objective To establish a simple and accurate method for determination of total polyphenols and vitamin C in fruits and vegetables. Methods Substances were extracted by solid-phase extraction column, and the water soluble interference reducing-substances were eluted by water, the polyphenols were eluted by methanol, Then the vitamin C were subtracted from water extract by the method of heating. The total polyphenols and vitamin C were determinate by correcting Folin-Cioculteu assay. Results The linear equation of this method was Y=0.0118X , R 2 = The condition of the Folin-Cioculteu colorimetry was confirmed, the maximum absorption wavelength is 760 nm, Folin-Cioculteu reagent concentration is 50%, 7.5% Na 2 CO 3 solution, 20~30, and stewing 60 min. The content of reducing substances of raw extract and water extract were determined by Folin-Cioculteu colorimetry, the total polyphenols content were assayed by subtracting the interfering reducing of water extract (WE) substances from the Folin-Cioculteu response of raw extract (RE). While the content of vitamin C was assayed by subtracting the Folin-Cioculteu response of heating water extraction from that of water extract (WE). Conclusion This method is simple and practical, the experiment condition is satisfied easily, the analysis results are reliable and that can be applied in the determination of total polyphenols and vitamin C. KEY WORDS: folin-cioculteu colorimetry; total polyphenols; vitamin C; solid-phase extraction * 通讯作者 :,,, mali0927@126.com *Corresponding author: MA Li, Engineer, Master, Food and Drug Control Institution of Hulun Buir, Hulun Buir , China. mali0927@126.com

2 4, : C 引言,,, [1,2], 1 g, C E [3], [4],, [5-8], [9],, :,,, [10-13],,,, C, [14-15],,,,,, HPLC HPLC C,,, ;,, HPLC, ; [16],, C,,,, C,,, C,,, 2 材料与方法 2.1 材料与试剂, 4 ; Sigma ; ; ; SPE(Oasis HLB, Waters); Milli-Q 2.2 仪器设备 UV-2450 (); (); BT125D (0.01 mg, Sartorius, ); XS365M (0.001 g, Precisa, ); (IKA, ); (IKA, ); ; ; ; LC-20A -() 2.3 试验方法原料提取物的制备 (RE) ()10 g,, 50 ml : (7: 3), 30 min, 50 ml 固相萃取分离多酚和水溶性成分 2 ml RE, HLB, (2 2 ml), (WE), (2 2 ml), 加热去除水溶性成分中的维生素 (HWE) (WE)85 2 h, C (HWE) 福林酚比色法条件的优化, 2 ml,, 1 ml, 2.5 ml, 10 ml,, SPSS17.0, FC 标准曲线的绘制 nm 1.0 mg/ml, μg/ml, nm 1.0 mg/ml,

3 μg/ml, 各多酚固相萃取回收率测定 , 1 ml SPE, (WE) : LC-20A DAD; : Diamonsil C 18, mm 5 μm; 1.0 ml/min; : A:0.5%, B:; : 0 10 min A:B 90%:10%, min A:B 50%:50%, min, B 100% 样品中总多酚和维生素含量 : C m V1 M V2 V3 M g C (mg/100 g); C --(μg/ml); m -- (g); V1 -- (ml); V2 -- (ml); V3 -- (ml); : (mg/100 g), (WE) (RE) : (mg/100 g)(hwe) (WE) 3 结果与分析 3.1 福林酚比色法条件优化,, W 6 W 5,, 760 nm 1~4, (P<0.05),, : (1: 1),, Na 2 CO 3, 7.5%, min, 90 min 120 min, 60 min, 表 1 福林酚试剂浓度对试验结果的影响 (n=3) Table 1 Effects of folin-cioculteu reagent concentration on the result of the measurement (n=3) 95% FC: N F Sig. 1: : : : : Table 2 表 2 NaCO 3 浓度因素对试验结果的影响 (n=3) Effects of NaCO 3 concentration on the result of the measurement (n=3) % N 95% F Sig

4 4, : C 1495 Table 3 表 3 温度因素对试验结果的影响 (n=3) Effects of treatment temperature on the result of the measurement (n=3) N 95% F Sig Table 4 表 4 反应时间因素对试验结果的影响 (n=3) Effects of response time on the result of the measurement (n=3) N 95% F Sig 标准曲线测定 FC 760 nm Y=0.0118X , R 2 =0.9989, Y=0.0095X , R 2 =0.9992,, 10~60 μg/ml, 80% 3.3 多酚回收率测定 Oasis HLB, HPLC,,, 5, 99%,, 23% 35%,,, 70%~90% 3.4 总多酚含量测定, Oasis HLB, 2 2 ml, WE, RE WE, HPLC WE, 3.5 维生素 C 含量测定 85 2 h C, HWE WE, C 3.6 FC 比色法在测定果蔬中总多酚和维生素 C 含量方面的应用, 6 1, 32.1%~48.6%,, 77.3% 72.6%, C C, 32% 43%, C 10%

5 表 5 不同多酚固相萃取的回收率 %, 以没食子酸计 (μg, n=3) Table 5 The recovery percents of different polyphenols by solid-phase extraction% express in Gallic acid(μg, n=3) 表 6 不同果蔬中总多酚和维生素的含量 (mg/100 g, n=3) Table 6 Content of total polyphenols and vitamin C in different fruits and vegetables(mg/100 g, n=3) a C a C b : a, b (n=3) PS: a expression in Gallic acid; b expression in ascorbic acid (n=3) 4 讨论 C, C,,,,,,,,,, C,, C, C,,, C,,,,,,, C,, C, 参考文献 [1] Ignat I, Volf I, Popa VI. A critical review of methods for characterization of polyphenolic compounds in fruits and vegetables [J]. Food Chem, 2011, (126): [2] Scalbert A, Williamson G. Dietary intake and bioavailability of polyphenols [J]. J Nutr, 2000, 130 (8S Suppl): 2073S 85S. 1 (, mg/100 g, n=3) Fig. 1 Comparison of total polyphenols and interference substances in different fruits and vegetables (express in Gallic acid, mg/100 g, n=3) [3] Beecher GR. Overview of dietary flavonoids nomenclature, occurrences and intake [J]. J Nutr, 2003, 133(10): 3248S 3254S. [4] Steinmetz KA, Potter JD. Vegetables, fruit, and cancer prevention: a review [J]. J Am Diet Assoc, 1996, (96): [5] Jansen MC, Mc Kenna D, Bueno DM, et al. Quantity and variety

4, : C 1497 of fruit and vegetable consumption and cancer risk [J]. Nutr Cancer, 2004, (48): 142 148. [6] Lamartiniere CA. Protection against breast cancer with genistein: a component of soy [J].

Wang PH, Zhao DW. Application of tea polyphenols in the food industry [J]. J Anhui Agric Sci, 2010, 38(33): 19075 19076, 19089. [9],,,. [J]., 2012, (S1): 110 117. Ying L, Jiang XC, Yao B, et al.

Determination of polyphenol in carambola by folin ciocalteu colorimetry [J]. Hunan Agric Sci, 2009, (7): 99 101. [11] Stephane G, Pierre B, Pascaline A, et al.

Spectrosc Spectr Anal, 2008, 28(7): 1630 1631. [13],,,. - [J]., 2014, 4(5): 1185 1190. Qin AX, Ji SJ, Gao Z, et al.

6 4, : C 1497 of fruit and vegetable consumption and cancer risk [J]. Nutr Cancer, 2004, (48): [6] Lamartiniere CA. Protection against breast cancer with genistein: a component of soy [J]. Am J Clin Nutr, 2000, (71): 1705S 1709S. [7] Senanayake SPJN. Green tea extract: chemistry, antioxidant properties and food applications a review [J]. J Funct Foods, 2013, 5(4): [8],. [J]., 2010, 38(33): , Wang PH, Zhao DW. Application of tea polyphenols in the food industry [J]. J Anhui Agric Sci, 2010, 38(33): , [9],,,. [J]., 2012, (S1): Ying L, Jiang XC, Yao B, et al. Research and application of antioxidant properties of lipid-soluble tea polyphenols [J]. China Food Addit, 2012, (S1): [10],,. Folin Ciocalteu [J]., 2009, (7): Lv QJ, Yi JR, Ding Y. Determination of polyphenol in carambola by folin ciocalteu colorimetry [J]. Hunan Agric Sci, 2009, (7): [11] Stephane G, Pierre B, Pascaline A, et al. Rapid determination of polyphenols and vitamin C in plant-derived products [J]. J Agric Food Chem, 2005, (53): [12]. [J]., 2008, 28(7): spectrophotometers [J]. Spectrosc Spectr Anal, 2008, 28(7): [13],,,. - [J]., 2014, 4(5): Qin AX, Ji SJ, Gao Z, et al. Determination of oil-soluble tea polyphenols in oil by solid phase extraction and Folin-Ciocalteu colorimetry [J]. J Food Saf Qual, 2014, 4(5): [14] Maria NI, Victoria FS, Costas GB, et al.development and validation of an HPLC-method for determination of free and bound phenolic acids in cereals after solid-phase extraction [J]. J Agric Food Chem, 2012, (134): [15],,. - [J]., 2009, (3)204: Weng HZ, Yue TX, Cheng YF. Solid-phase extraction and high-performance liquid chromatography determination of polyphenols in wine [J].China Brewing, 2009, 204(3): [16] David WJ, Mercurio MD, Markus JH, et al. Rapid isolation of red wine polymeric polyphenols by solid-phase extraction [J]. J Agric Food Chem, 2008, (56): 作者简介 ( 责任编辑 : 白洪健 ) 马丽, 质量检验工程师, 硕士, 主要研究方向为食品检验 mali0927@126.com Zhang SF. Determination of polyphenols in tea by a new method

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.