HPLC法测定李果实中花色苷含量及其与pH示差法的比较研究
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摘要
通过参数优化,建立了HPLC法同时测定李果实中5种花色苷(飞燕草素-3-葡萄糖苷、飞燕草素-3-芸香糖苷、矢车菊素-3-葡萄糖苷、矢车菊素-3-芸香糖苷、芍药素-3-葡萄糖苷)含量的方法,并比较了HPLC法与p H值示差法在测定李果实花色苷总量上的差异。结果表明:以乙腈-水(0.3%磷酸)为流动相,梯度洗脱,检测波长为510 nm,50 min内可实现5种花色苷的完全分离,线性范围为1~100μg·ml-1(R~2=0.9980~0.9997),加样回收率为97.34%~103.17%、RSD为0.83%~1.24%。HPLC法与p H值示差法测定李花色苷总含量的对比结果表明,两者间的差距与花色苷含量有关,花色苷含量越高,两者相差越大,且HPLC法的测定值较高;相反,花色苷含量越低,两者相差越小,且HPLC法的测定值较低。
A reverse ph ase HPL C method was utilized to investigate anthocyanin gategories( delphin-3-glucoside, delphin-3-richinoside, cyanidin-3-glucoside, cyanidin-3-richinoside and peonidin-3-glucoside) and content in fruits of different plum cultivars.The total anthocyanin content in plum fruit was measured by HPLC method and p H differential method, respectively and the differences between two methods were compared. The results showed that the anthocyanin gategories were thoroughly separated within 50 min under the condition of using acetonitrile-water(contained 0.3% phosphoric acid) as mobile phase with gradient elution at a detection wavelength of 510 nm. The method exhibited a linear range from 1 to 100 μg·ml-1( with the correlation coefficients ranged from0.9980 to 0.9997), the recovery rate was from 97.34% to 103.17%, and the RSD was 0.83% to 1.24%. There was remarkable difference between HPLC method and p H differential method for the determination of total content of anthocyanin. The results also showed that the difference was related to the content of anthocyanin. If a cultivar contained more anthocyanin, the value of HPLC method was much higher than that of p H differential method; on the contrary, if a cultivar contained litter anthocyanin, the values became close to each other, and even the value of HPLC method was less than that of p H differential method.
A reverse ph ase HPL C method was utilized to investigate anthocyanin gategories( delphin-3-glucoside, delphin-3-richinoside, cyanidin-3-glucoside, cyanidin-3-richinoside and peonidin-3-glucoside) and content in fruits of different plum cultivars.The total anthocyanin content in plum fruit was measured by HPLC method and p H differential method, respectively and the differences between two methods were compared. The results showed that the anthocyanin gategories were thoroughly separated within 50 min under the condition of using acetonitrile-water(contained 0.3% phosphoric acid) as mobile phase with gradient elution at a detection wavelength of 510 nm. The method exhibited a linear range from 1 to 100 μg·ml-1( with the correlation coefficients ranged from0.9980 to 0.9997), the recovery rate was from 97.34% to 103.17%, and the RSD was 0.83% to 1.24%. There was remarkable difference between HPLC method and p H differential method for the determination of total content of anthocyanin. The results also showed that the difference was related to the content of anthocyanin. If a cultivar contained more anthocyanin, the value of HPLC method was much higher than that of p H differential method; on the contrary, if a cultivar contained litter anthocyanin, the values became close to each other, and even the value of HPLC method was less than that of p H differential method.
引文
[1]周丹蓉,方智振,廖汝玉,等.李果皮花色素苷、类黄酮和类胡萝卜素含量及抗氧化性研究[J].营养学报,2013,35(6):571-576.
[2]周丹蓉,廖汝玉,叶新福.水果中主要功能性成分研究进展[J].福建农业学报,2011,26(6):1129-1134.
[3]Wu X,Cao G,Prior R L.Absorption and metabolism of anthocyanins in elderly women after consumption of elderberry or blueberry[J].The Journal of Nutrition,2002,132(7):1865-1871.
[4]郑永霞.花色素苷药理功效的研究进展[J].山西医药杂志:上半月,2008,37(3):255-257.
[5]Fossen T,Slimestad R,Andersen O M.Anthocyanins from maize(Zea mays)and reed canarygrass(Phalaris arundinacea)[J].Journal of Agricultural and Food Chemistry,2001,49(5):2318-2321.
[6]Chandra A,Rana J,Li Y.Separation,identification,quantification,and method validation of anthocyanins in botanical supplement raw materials by HPLC and HPLC-MS[J].Journal of Agricultural and Food Chemistry,2001,49(8):35l5-3521.
[7]Terahara N,Callebaut A,Ohba R,et a1.Acylated anthocyanidin 3-sophoroside-5-glucosides from Ajuga reptans flowers and the corresponding cell cultures[J].Phytochemisty,2001,58(3):493-500.
[8]张学英.李果实着色与花色素苷合成机理研究[D].杭州:浙江大学,2008.
[9]刘奕琳,王振宇.蓝靛果中花色苷含量的测定及其体外抗氧化性[J].中国林副特产,2011,114(5):14-17.
[10]杨桂霞,范海林,李亚东,等.RP-HPLC法测定栽培种越橘果中花色素苷的含量[J].药物分析杂志,2005,25(10):1222-1224.
[11]García-B E,Cabello F,Revilla E.Analysis of grape and wine anthocyanins by HPLC-MS[J].Journal of Agricultural and Food Chemistry,2003,51(19):5622-5629.
[12]Qlngguo T,Izabela K,Steven J S.Probing Anthocyanin Profiles in Purple Sweet Potato Cell Line(Ipomoea batatas L.Cv.Ayamurasaki)by High-Performance Liquid Chromatography and Electrospray lonization Tandem Mass Spectrometry[J].Journal of Agricultural and Food Chemistry,2005,53(16):6503-6509.
[13]刘宽博,王明力,万良钰,等.树莓中主要活性成分及产品研究进展[J].中国南方果树,2016,45(6):178-183.
[14]Francisco A T,Maria I G,Paedar C,et al.HPLC-DAD-ESIMS analysis of phenolic compound in nectarine,peaches and plum[J].Journal of Agricultural and Food Chemistry,2001,49(10):4748-4760.
[15]张元慧.李(Prunus spp.)果实色泽发育机理研究[D].保定:河北农业大学,2003.
[16]Fuleki T,Francis F J.Quantitative methods for anthocyanins.Extraction and determination of total anthocyanin incranberries[J].Journal of Food Science,1968,33(1):72-77.
[17]胡桂兵,陈大成,李平,等.荔枝果皮色素、酚类物质与酶活性的动态变化[J].果树学报,2000,17(1):35-40.
[18]王少敏,高华君,刘嘉芬,等.套袋短枝红富士果实内含物及果皮色素的变化[J].果树科学,2000,17(1):76-77.
[2]周丹蓉,廖汝玉,叶新福.水果中主要功能性成分研究进展[J].福建农业学报,2011,26(6):1129-1134.
[3]Wu X,Cao G,Prior R L.Absorption and metabolism of anthocyanins in elderly women after consumption of elderberry or blueberry[J].The Journal of Nutrition,2002,132(7):1865-1871.
[4]郑永霞.花色素苷药理功效的研究进展[J].山西医药杂志:上半月,2008,37(3):255-257.
[5]Fossen T,Slimestad R,Andersen O M.Anthocyanins from maize(Zea mays)and reed canarygrass(Phalaris arundinacea)[J].Journal of Agricultural and Food Chemistry,2001,49(5):2318-2321.
[6]Chandra A,Rana J,Li Y.Separation,identification,quantification,and method validation of anthocyanins in botanical supplement raw materials by HPLC and HPLC-MS[J].Journal of Agricultural and Food Chemistry,2001,49(8):35l5-3521.
[7]Terahara N,Callebaut A,Ohba R,et a1.Acylated anthocyanidin 3-sophoroside-5-glucosides from Ajuga reptans flowers and the corresponding cell cultures[J].Phytochemisty,2001,58(3):493-500.
[8]张学英.李果实着色与花色素苷合成机理研究[D].杭州:浙江大学,2008.
[9]刘奕琳,王振宇.蓝靛果中花色苷含量的测定及其体外抗氧化性[J].中国林副特产,2011,114(5):14-17.
[10]杨桂霞,范海林,李亚东,等.RP-HPLC法测定栽培种越橘果中花色素苷的含量[J].药物分析杂志,2005,25(10):1222-1224.
[11]García-B E,Cabello F,Revilla E.Analysis of grape and wine anthocyanins by HPLC-MS[J].Journal of Agricultural and Food Chemistry,2003,51(19):5622-5629.
[12]Qlngguo T,Izabela K,Steven J S.Probing Anthocyanin Profiles in Purple Sweet Potato Cell Line(Ipomoea batatas L.Cv.Ayamurasaki)by High-Performance Liquid Chromatography and Electrospray lonization Tandem Mass Spectrometry[J].Journal of Agricultural and Food Chemistry,2005,53(16):6503-6509.
[13]刘宽博,王明力,万良钰,等.树莓中主要活性成分及产品研究进展[J].中国南方果树,2016,45(6):178-183.
[14]Francisco A T,Maria I G,Paedar C,et al.HPLC-DAD-ESIMS analysis of phenolic compound in nectarine,peaches and plum[J].Journal of Agricultural and Food Chemistry,2001,49(10):4748-4760.
[15]张元慧.李(Prunus spp.)果实色泽发育机理研究[D].保定:河北农业大学,2003.
[16]Fuleki T,Francis F J.Quantitative methods for anthocyanins.Extraction and determination of total anthocyanin incranberries[J].Journal of Food Science,1968,33(1):72-77.
[17]胡桂兵,陈大成,李平,等.荔枝果皮色素、酚类物质与酶活性的动态变化[J].果树学报,2000,17(1):35-40.
[18]王少敏,高华君,刘嘉芬,等.套袋短枝红富士果实内含物及果皮色素的变化[J].果树科学,2000,17(1):76-77.