蓝果忍冬酚类物质的提取、鉴定及抗氧化性研究
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摘要
本文以蓝果忍冬主要栽培品种贝瑞尔为试验材料,对其叶片和果实中所含的酚类物质进行提取工艺研究,分析了不同生长发育时期其叶片和果实酚类物质含量的变化趋势。并以贝瑞尔、蓝鸟、蓝纺锤三个蓝果忍冬栽培品种和蓝靛果忍冬、巴利斯忍冬、博奇卡勒尼卡娃忍冬、阿尔泰忍冬、库页岛忍冬五个野生蓝果忍冬种类为试验材料,对其叶片和果实所含的酚类物质进行含量测定,并鉴定了不同种类及品种蓝果忍冬果实中酚类物质的结构,分析了其叶片及果实酚类物质提取液的体外抗氧化能力。
在蓝果忍冬叶片和果实酚类物质的提取过程中,以贝瑞尔为试验材料,采用乙醇浸提法进行了提取条件的优化试验,对影响提取效果的四个因素,包括乙醇浓度、乙醇体积、提取温度和提取时间进行了单因素试验,并在单因素的基础上进行正交试验设计,确定了蓝果忍冬叶片和果实酚类物质提取的最佳工艺。
采用HPLC/MS方法鉴定了八个不同品种及种类的蓝果忍冬果实花色苷的结构及其含量。
分别采用铁离子还原能力测定、抗脂质过氧化能力测定、清除超氧阴离子自由基能力测定、清除羟自由基能力测定、DPPH清除试验、ABTS自由基清除试验六种方法,分析了蓝果忍冬叶片及果实的体外抗氧化能力。
试验的主要结果如下:
1.通过对影响乙醇浸提效果的主要因素进行单因素和正交试验,分别确定了蓝果忍冬叶片和果实酚类物质提取的最优条件。叶片酚类物质提取的最佳条件为:乙醇浓度50%、乙醇体积50 mL、提取温度50℃,提取时间2.5 h。此提取条件下蓝果忍冬叶片酚类物质的含量为14.14 mg/g。果实酚类物质提取的最佳条件为:乙醇浓度70 %、乙醇体积50 mL、提取温度50℃、提取时间3.0 h。此提取条件下蓝果忍冬果实酚类物质含量为24.44 mg/g。
2.不同生长发育时期蓝果忍冬叶片酚类物质含量分别为:未坐果期14.13 mg/g、绿果期12.56 mg/g、开始着色期8.99 mg/g、部分着色期7.64 mg/g、完全着色期7.42 mg/g。蓝果忍冬果实酚类物质含量分别为:绿果期10.34 mg/g、开始着色期11.86 mg/g、部分着色期15.39 mg/g、完全着色期24.45 mg/g。
3.不同种类及品种的蓝果忍冬叶片酚类物质含量为:贝瑞尔13.63 mg/g、蓝鸟9.36 mg/g、蓝纺锤17.10 mg/g、蓝靛果忍冬12.78 mg/g、巴利斯忍冬12.66 mg/g、博奇卡勒尼卡娃忍冬13.87 mg/g、阿尔泰忍冬15.58 mg/g、库页岛忍冬3.00 mg/g。果实酚类物质含量为:贝瑞尔24.19 mg/g、蓝鸟23.02 mg/g、蓝纺锤26.33 mg/g、蓝靛果忍冬24.12 mg/g、巴里斯忍冬24.17 mg/g、博奇卡勒尼卡娃忍冬25.24 mg/g、阿尔泰忍冬27.43 mg/g、库页岛忍冬22.98 mg/g。
4.鉴定出蓝果忍冬果实中主要含有八种花色苷,分别为:矢车菊3,5-二葡萄糖苷,矢车菊-3-葡萄糖苷,矢车菊-芸香糖苷,天竺葵-3-葡萄糖苷,芍药-3-葡萄糖苷,芍药-3-芸香糖苷,矢车菊-(丙酮基)-己糖苷以及矢车菊衍生物。
5.通过各种抗氧化能力分析方法对蓝果忍冬的抗氧化能力进行了评价:
铁离子还原能力的强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。
抗脂质过氧化能力强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。其中阿尔泰忍冬、博奇卡勒尼卡娃忍冬抗脂质过氧化能力接近合成抗氧化剂BHT的抗脂质过氧化能力。
清除超氧阴离子自由基能力强弱顺序为:蓝靛果忍冬>贝瑞尔>蓝纺锤>巴利斯忍冬>阿尔泰忍冬>蓝鸟>博奇卡勒尼卡娃忍冬>库页岛忍冬。
清除羟自由基能力的强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。
清除DPPH自由基能力的强弱顺序为:蓝纺锤>蓝鸟>巴利斯忍冬>贝瑞尔>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>阿尔泰忍冬>库页岛忍冬。
清除ABTS自由基能力强弱顺序为:蓝纺锤>蓝鸟>巴利斯忍冬>贝瑞尔>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>阿尔泰忍冬>库页岛忍冬。
In this paper, the L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica which is the main cultivars of Blue honeysuckle were used as experimental material to research the extraction technology of phenols in its leaves and berries,and analysis the trend of phenols in different periods.Three cultivars including L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica, Blue bird(L.caerulea subsp.Kamtschatica) and Blue spindle (L.caerulea ubsp.Kamtschaticas), and five wild species including (L.caerulea var. edulis),(L.caerulea subsp.pallasii),(L.boczkarnikowii), (L.caerulea subsp.altaicai)and(L.caerulea subsp. emphyllocalyx) as the test material to determine the content of phenols in their leaves and berries,and identify the compone- nts in different speci- es and varieties of Blue honeysuckle,At last,analysis the vitro antioxidant capacity of leaves and berries.
During the extraction to phenols in leaf and berries of Blue honeysuckle, the L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica which is the main cultivars of Blue honeysuckle were used as experimental material to screen the optimal conditions using ethanol extraction met- hod. Consider the four influencing factors, including the ethanol concentration, ethanol volume, extracting temperature and extracting time for a single-factor test, and basing on single-factor made orthogonal experimental design to choose the optimum extraction process of phenols in leaves and berries of Blue honeysuckle.
Using six methods, including ferric reducing antioxidant power, fraction with anti-lipid per- oxidation, superoxide anion radical scavenging activity, hydroxyl free radical scavenging activity, DPPH radical scavenging activity, ABTS radical scavenging activity, analyze the antioxidant cap- acity of Blue honeysuckle.
The study's major findings are as follows:
1. Through the single factor and orthogonal tests of the main influencing factors were determed- ined the optimal conditions for phenolic extraction. The best conditions of phenolic extracting from leaves were: ethanol concentration 50 %, ethanol volume 50 mL, extracting temp- erature 50℃, extracting time 2.5 h. In this extracting conditions, the phenolic content in leaves of Blue honeys- honeysuckle were 14.14 mg/g. The best extraction conditions of berries were: ethanol concentration 70, ethanol volum 50 mL, extracting temperature 50℃, extracting time 3.0 h. In this extraction conditions, the phenolic content were 24.44 mg/g.
2. The phenolic contents of leaves in different periods were: the period of no beries 14.13 mg/g, the period of green berries 12.56 mg/g, the period of started coloring 8.99 mg/g, the period of parted coloring 7.64 mg/g, the period of full coloring 7.42 mg/g. The phenlic contents of berri- es in different periods were: the period of green berries 10.34 mg/g, the period of started coloring 11.86 mg/g, the period of parted coloring 15.39 mg/g, the period of full coloring 24.45 mg / g.
3. The phenolic contents of leaves in different species and varieties of Blue honeysuckle were: L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica 13.63 mg/g, Blue bird (L.caerulea subsp.Kamtschatica) 9.36 mg/g, Blue spindle (L.caerulea ubsp.Kamtschaticas) 17.10 mg/g, (L.caerulea var. edulis) 12.78 mg/g, (L.caerulea subsp.pallasii) 12.66 mg/g, (L.boczkarnikowii) 13.87 mg/g, (L.caerulea subsp.altaicai) 15.58 mg / g, (L.caerulea subsp. emphyllocalyx) 3.00 mg / g.And the phenolic contents of berries were: L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica 24.19 mg / g, Blue bird (L.caerulea subsp.Kamtschatica) 23.02 mg / g, (L.caerulea subsp.pallasii) 26.33 mg / g, (L.caerulea var. edulis) 24.12 mg / g, (L.caerulea subsp.pallasii) 24.17 mg / g, (L.boczkarnikowii) 25.24 mg / g, (L.caerulea subsp.altaicai) 27.43 mg / g, (L.caerulea subsp. emphyllocalyx) 22.98 mg / g.
4. Eight major anthocyanins are identification in berries of Blue honeysuckles, including cyanidin 3, 5-diglucoside, cyanidin 3-glucoside, cyanidin 3-rutinoside, pelargonidin 3-glucoside, pelargonidin 3-glucosid, peonidin 3-glucoside, peonidin 3-rutinoside, cyaniding- (acetoyl)-hexosi- de and cyanidin-derivatives.
5. Antioxidant capacity through various analytical methods blue Lonicera antioxidant capacity was evaluated:
Ferric reducing ability of the strength of the order: (L.caerulea subsp.altaicai)> (L.caerulea subsp.pallasii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)>Bluebird>L.cae- rulea subsp.altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. Emphyllo- calyx).
Anti-lipid peroxidation descending order: (L.caerulea subsp.altaicai) > (L.caerulea subsp.pal- lasii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)> Bluebird> L.caerulea sub- sp.altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. emphyllocalyx).
Superoxide anion radicals in descending order: L.caerulea var. edulis)>(L.caerulea subsp.alt- aicai×L.caerulea subsp.Kamtschatica)> Blue spindle > (L.caerulea subsp.pallasii)> (L.caerulea subsp.altaicai)> Bluebird >(L.boczkarnikowii)> (L.caerulea subsp. emphyllocalyx)
Scavenging ability in descending order: (L.caerulea subsp.altaicai) > (L.caerulea subsp.pall- asii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)> Bluebird> L.caerulea subsp. altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. emphyllocalyx).
DPPH free radical ability to clear the strength of the order: Blue spindle >Bluebird >(L.cae- rulea subsp.pallasii)> (L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica)> (L.boczkami- kowii)>(L.caerulea var. edulis)>(L.caerulea subsp.altaicai)> (L.caerulea subsp. emphyllocalyx).
Ability to remove ABTS radical in descending order: Blue spindle > Bluebird >(L.caerulea subsp.pallasii)> (L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica)> (L.boczkarmikow- ii)>(L.caerulea var. edulis)>(L.caerulea subsp.altaicai)> (L.caerulea subsp. emphyllocalyx).
在蓝果忍冬叶片和果实酚类物质的提取过程中,以贝瑞尔为试验材料,采用乙醇浸提法进行了提取条件的优化试验,对影响提取效果的四个因素,包括乙醇浓度、乙醇体积、提取温度和提取时间进行了单因素试验,并在单因素的基础上进行正交试验设计,确定了蓝果忍冬叶片和果实酚类物质提取的最佳工艺。
采用HPLC/MS方法鉴定了八个不同品种及种类的蓝果忍冬果实花色苷的结构及其含量。
分别采用铁离子还原能力测定、抗脂质过氧化能力测定、清除超氧阴离子自由基能力测定、清除羟自由基能力测定、DPPH清除试验、ABTS自由基清除试验六种方法,分析了蓝果忍冬叶片及果实的体外抗氧化能力。
试验的主要结果如下:
1.通过对影响乙醇浸提效果的主要因素进行单因素和正交试验,分别确定了蓝果忍冬叶片和果实酚类物质提取的最优条件。叶片酚类物质提取的最佳条件为:乙醇浓度50%、乙醇体积50 mL、提取温度50℃,提取时间2.5 h。此提取条件下蓝果忍冬叶片酚类物质的含量为14.14 mg/g。果实酚类物质提取的最佳条件为:乙醇浓度70 %、乙醇体积50 mL、提取温度50℃、提取时间3.0 h。此提取条件下蓝果忍冬果实酚类物质含量为24.44 mg/g。
2.不同生长发育时期蓝果忍冬叶片酚类物质含量分别为:未坐果期14.13 mg/g、绿果期12.56 mg/g、开始着色期8.99 mg/g、部分着色期7.64 mg/g、完全着色期7.42 mg/g。蓝果忍冬果实酚类物质含量分别为:绿果期10.34 mg/g、开始着色期11.86 mg/g、部分着色期15.39 mg/g、完全着色期24.45 mg/g。
3.不同种类及品种的蓝果忍冬叶片酚类物质含量为:贝瑞尔13.63 mg/g、蓝鸟9.36 mg/g、蓝纺锤17.10 mg/g、蓝靛果忍冬12.78 mg/g、巴利斯忍冬12.66 mg/g、博奇卡勒尼卡娃忍冬13.87 mg/g、阿尔泰忍冬15.58 mg/g、库页岛忍冬3.00 mg/g。果实酚类物质含量为:贝瑞尔24.19 mg/g、蓝鸟23.02 mg/g、蓝纺锤26.33 mg/g、蓝靛果忍冬24.12 mg/g、巴里斯忍冬24.17 mg/g、博奇卡勒尼卡娃忍冬25.24 mg/g、阿尔泰忍冬27.43 mg/g、库页岛忍冬22.98 mg/g。
4.鉴定出蓝果忍冬果实中主要含有八种花色苷,分别为:矢车菊3,5-二葡萄糖苷,矢车菊-3-葡萄糖苷,矢车菊-芸香糖苷,天竺葵-3-葡萄糖苷,芍药-3-葡萄糖苷,芍药-3-芸香糖苷,矢车菊-(丙酮基)-己糖苷以及矢车菊衍生物。
5.通过各种抗氧化能力分析方法对蓝果忍冬的抗氧化能力进行了评价:
铁离子还原能力的强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。
抗脂质过氧化能力强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。其中阿尔泰忍冬、博奇卡勒尼卡娃忍冬抗脂质过氧化能力接近合成抗氧化剂BHT的抗脂质过氧化能力。
清除超氧阴离子自由基能力强弱顺序为:蓝靛果忍冬>贝瑞尔>蓝纺锤>巴利斯忍冬>阿尔泰忍冬>蓝鸟>博奇卡勒尼卡娃忍冬>库页岛忍冬。
清除羟自由基能力的强弱顺序为:阿尔泰忍冬>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>巴利斯忍冬>蓝鸟>贝瑞尔>蓝纺锤>库页岛忍冬。
清除DPPH自由基能力的强弱顺序为:蓝纺锤>蓝鸟>巴利斯忍冬>贝瑞尔>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>阿尔泰忍冬>库页岛忍冬。
清除ABTS自由基能力强弱顺序为:蓝纺锤>蓝鸟>巴利斯忍冬>贝瑞尔>博奇卡勒尼卡娃忍冬>蓝靛果忍冬>阿尔泰忍冬>库页岛忍冬。
In this paper, the L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica which is the main cultivars of Blue honeysuckle were used as experimental material to research the extraction technology of phenols in its leaves and berries,and analysis the trend of phenols in different periods.Three cultivars including L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica, Blue bird(L.caerulea subsp.Kamtschatica) and Blue spindle (L.caerulea ubsp.Kamtschaticas), and five wild species including (L.caerulea var. edulis),(L.caerulea subsp.pallasii),(L.boczkarnikowii), (L.caerulea subsp.altaicai)and(L.caerulea subsp. emphyllocalyx) as the test material to determine the content of phenols in their leaves and berries,and identify the compone- nts in different speci- es and varieties of Blue honeysuckle,At last,analysis the vitro antioxidant capacity of leaves and berries.
During the extraction to phenols in leaf and berries of Blue honeysuckle, the L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica which is the main cultivars of Blue honeysuckle were used as experimental material to screen the optimal conditions using ethanol extraction met- hod. Consider the four influencing factors, including the ethanol concentration, ethanol volume, extracting temperature and extracting time for a single-factor test, and basing on single-factor made orthogonal experimental design to choose the optimum extraction process of phenols in leaves and berries of Blue honeysuckle.
Using six methods, including ferric reducing antioxidant power, fraction with anti-lipid per- oxidation, superoxide anion radical scavenging activity, hydroxyl free radical scavenging activity, DPPH radical scavenging activity, ABTS radical scavenging activity, analyze the antioxidant cap- acity of Blue honeysuckle.
The study's major findings are as follows:
1. Through the single factor and orthogonal tests of the main influencing factors were determed- ined the optimal conditions for phenolic extraction. The best conditions of phenolic extracting from leaves were: ethanol concentration 50 %, ethanol volume 50 mL, extracting temp- erature 50℃, extracting time 2.5 h. In this extracting conditions, the phenolic content in leaves of Blue honeys- honeysuckle were 14.14 mg/g. The best extraction conditions of berries were: ethanol concentration 70, ethanol volum 50 mL, extracting temperature 50℃, extracting time 3.0 h. In this extraction conditions, the phenolic content were 24.44 mg/g.
2. The phenolic contents of leaves in different periods were: the period of no beries 14.13 mg/g, the period of green berries 12.56 mg/g, the period of started coloring 8.99 mg/g, the period of parted coloring 7.64 mg/g, the period of full coloring 7.42 mg/g. The phenlic contents of berri- es in different periods were: the period of green berries 10.34 mg/g, the period of started coloring 11.86 mg/g, the period of parted coloring 15.39 mg/g, the period of full coloring 24.45 mg / g.
3. The phenolic contents of leaves in different species and varieties of Blue honeysuckle were: L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica 13.63 mg/g, Blue bird (L.caerulea subsp.Kamtschatica) 9.36 mg/g, Blue spindle (L.caerulea ubsp.Kamtschaticas) 17.10 mg/g, (L.caerulea var. edulis) 12.78 mg/g, (L.caerulea subsp.pallasii) 12.66 mg/g, (L.boczkarnikowii) 13.87 mg/g, (L.caerulea subsp.altaicai) 15.58 mg / g, (L.caerulea subsp. emphyllocalyx) 3.00 mg / g.And the phenolic contents of berries were: L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica 24.19 mg / g, Blue bird (L.caerulea subsp.Kamtschatica) 23.02 mg / g, (L.caerulea subsp.pallasii) 26.33 mg / g, (L.caerulea var. edulis) 24.12 mg / g, (L.caerulea subsp.pallasii) 24.17 mg / g, (L.boczkarnikowii) 25.24 mg / g, (L.caerulea subsp.altaicai) 27.43 mg / g, (L.caerulea subsp. emphyllocalyx) 22.98 mg / g.
4. Eight major anthocyanins are identification in berries of Blue honeysuckles, including cyanidin 3, 5-diglucoside, cyanidin 3-glucoside, cyanidin 3-rutinoside, pelargonidin 3-glucoside, pelargonidin 3-glucosid, peonidin 3-glucoside, peonidin 3-rutinoside, cyaniding- (acetoyl)-hexosi- de and cyanidin-derivatives.
5. Antioxidant capacity through various analytical methods blue Lonicera antioxidant capacity was evaluated:
Ferric reducing ability of the strength of the order: (L.caerulea subsp.altaicai)> (L.caerulea subsp.pallasii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)>Bluebird>L.cae- rulea subsp.altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. Emphyllo- calyx).
Anti-lipid peroxidation descending order: (L.caerulea subsp.altaicai) > (L.caerulea subsp.pal- lasii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)> Bluebird> L.caerulea sub- sp.altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. emphyllocalyx).
Superoxide anion radicals in descending order: L.caerulea var. edulis)>(L.caerulea subsp.alt- aicai×L.caerulea subsp.Kamtschatica)> Blue spindle > (L.caerulea subsp.pallasii)> (L.caerulea subsp.altaicai)> Bluebird >(L.boczkarnikowii)> (L.caerulea subsp. emphyllocalyx)
Scavenging ability in descending order: (L.caerulea subsp.altaicai) > (L.caerulea subsp.pall- asii)> (L.caerulea subsp.pallasii)> (L.caerulea subsp. emphyllocalyx)> Bluebird> L.caerulea subsp. altaicai×L.caerulea subsp.Kamtschatica > Blue spindle> (L.caerulea subsp. emphyllocalyx).
DPPH free radical ability to clear the strength of the order: Blue spindle >Bluebird >(L.cae- rulea subsp.pallasii)> (L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica)> (L.boczkami- kowii)>(L.caerulea var. edulis)>(L.caerulea subsp.altaicai)> (L.caerulea subsp. emphyllocalyx).
Ability to remove ABTS radical in descending order: Blue spindle > Bluebird >(L.caerulea subsp.pallasii)> (L.caerulea subsp.altaicai×L.caerulea subsp.Kamtschatica)> (L.boczkarmikow- ii)>(L.caerulea var. edulis)>(L.caerulea subsp.altaicai)> (L.caerulea subsp. emphyllocalyx).
引文
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