油溶性茶多酚含量的测定 
Assay Test for Green Tea Polyphenol Oil Soluble 

A.4.1试剂和材料 
Preparation 
A.4.1.1油溶性茶多酚：经45 ℃真空干燥至恒重后使用。 
Green tea polyphenol oil soluble: at 45 ℃ Vacuum drying until constant weight 
 
A.4.1.2维生素C：经45 ℃真空干燥至恒重后使用。 
Vitamin C: at 45 ℃ Vacuum drying until constant weight 
A.4.1.3没食子酸乙酯：样品经重结晶纯化，高效液相色谱检测纯度99.9%。经45 ℃真空干燥至恒重后使用。 
ethyl gallate:Recrystallization purification,HPLC 99.9%,at 45 ℃ Vacuum drying until constant weight 
A.4.1.4 维生素C溶液：0.075 mol/L，准确称取1.3222 g维生素C，用水溶解后定容至100 mL。 
Vitamin C solution: 0.075 mol/L, take 1.3222 g of Vitamin C, dissolve it in the water to 100 mL 
A.4.1.5 三乙醇胺溶液：量取50.0 mL三乙醇胺，加入25 mL水稀释后转入100 mL容量瓶中，并用水稀释至刻度。 
triethanolamine solution:take 50.0 mL of triethanolamine,add 25 mL water,put into 100 mL of measuring flask,add water to the scale 
 
A.4.1.6 没食子酸乙酯标准储备溶液：准确称取0.1000 g没食子酸乙酯，用水溶解后定容至100 mL。 
ethyl gallate solution:take 0.1000 g of ethyl gallate,dissolve it in the water to 100 mL 
A.4.1.7 油溶性茶多酚溶液：准确称取约500 mg油溶性茶多酚（精确至0. 1 mg），记为油溶性茶多酚干重m，用无水乙醇溶解并定容至100 mL。 
Green tea polyphenol oil soluble solution:take 500 mg(accurate to 0.1 mg),mark dry weight m of Green tea polyphenol oil soluble,dissolve it in the absolute ethyl alcohol to 100 mL 
A.4.1.8 酒石酸亚铁溶液：准确称取1.6050 g硫酸亚铁（FeSO4·7H2O）和8.0440 g酒石酸钾钠，分别用水溶解后转入至1000 mL容量瓶中，稀释至刻度。 
Ferrous tartrate solution:take 1.6050 g of FeSO4·7H2O and 8.0440 g of Sodium potassium tartrate,dissolve them in the water respectively,then to 1000mL,add water to the scale 

A.4.2分析步骤 
Analysis procedure 
A.4.2.1 于25 mL容量瓶中依次移入5 mL酒石酸亚铁溶液﹑1 mL三乙醇胺溶液、2 mL维生素C溶液，用水定容至刻度，充分摇匀。此为空白溶液（1）。 
put 5 mL of Ferrous tartrate solution,1 mL of triethanolamine solution and 2 mL of Vitamin C solution into 25 mL of measuring flask,add water to the scale,shake well,get solution No.(1) 
A.4.2.2 取5个25 mL容量瓶分别依次加入5 mL酒石酸亚铁溶液﹑1 mL三乙醇胺溶液、2 mL维生素C溶液，然后用移液管分别移取0.2 mL、0.4 mL、0.6 mL、0.8 mL、1.0 mL的没食子酸乙酯标准储备溶液加到容量瓶中，以水定容至刻度，充分摇匀。配制好的溶液在室温下放置1 h。以空白溶液（1）为参照，在520 nm波长下测定该溶液的吸光度，以没食子酸乙酯标准溶液浓度对溶液吸光度建立没食子酸乙酯标准曲线(标准曲线线性相关系数要求0.999以上)。 
take 5 pieces of 25 mL's measuring flask,each one is added by 5 mL of Ferrous tartrate solution,1 mL of triethanolamine solution and 2 mL of Vitamin C solution 
take 0.2 mL,0.4 mL,0.6 mL,0.8 mL and 1.0 mL of ethyl gallate solution by pipette respectively,add them into above 5 solutions separately,then each one is added water to the scale and shake well,place these for 1 hour at room temperature 
take solution No.(1) as the reference, test the absorbance of solution at 520 nm of wavelengths,the standard curve of ethyl gallate is established with its solution and the absorbance (linear correlation coefficient is above 0.999) 
A.4.2.3 于25 mL容量瓶中依次移入5 mL酒石酸亚铁溶液﹑1 mL三乙醇胺溶液、2 mL维生素C溶液，移入1 mL（记为体积v）的油溶性茶多酚溶液（放液速度要慢），用少许无水乙醇吹洗瓶口的残留液至容量瓶，以水定容至刻度，充分摇匀。配制好的溶液在室温下放置1 h，以空白溶液（1）为参照，在520 nm波长下测定该溶液吸光度。所得吸光度在没食子酸乙酯标准曲线上查到对应的浓度c（μg/mL），该浓度乘以换算系数3.5，即为测得的油溶性茶多酚浓度。 
add 5 mL of Ferrous tartrate solution,1 mL of triethanolamine solution and 2 mL of Vitamin C solution into 25 mL's measuring flask 
add 1 mL (mark v) of Green tea polyphenol oil soluble solution slowly 
wash the residual liquid in the measuring flask mouth into the measuring flask by a little absolute ethyl alcohol solution 
add water to the scale,shake well,place these for 1 hour at room temperature 
take solution No.(1) as the reference,test the absorbance of solution at 520 nm of wavelengths,the test result of absorbance is found at the corresponding concentration of c (μg/mL) on the standard curve of ethyl gallate,this *3.5,get the assay 
 
油溶性茶多酚含量的质量分数w，按公式（A.1）计算： 
                …………(A.1) 
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