CN115044407B - 一种白背叶楤木挥发油及其制备方法与应用 - Google Patents
一种白背叶楤木挥发油及其制备方法与应用 Download PDFInfo
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Abstract
本发明公开了一种白背叶楤木挥发油及其制备方法与应用，所述白背叶楤木挥发油以[1aR‑(1aα,4α,7β,7bα)]‑1a,2,3,4,4a,5,6,7b‑十氢‑1,1,7‑三甲基‑4‑亚甲基‑1H‑环丙烯并[E]薁‑7‑醇、植酮、棕榈酸甲酯、叶绿醇和α‑松油醇为主要成分；其制备方法是以白背叶楤木为原料，粉碎后加水浸泡后用水蒸气蒸馏法提取，去掉水层，用有机溶剂反复洗涤提取器，得白背叶楤木挥发油粗提物，最后经无水硫酸钠干燥，过滤，滤液用旋转蒸发仪将有机溶剂挥干得到目标白背叶楤木挥发油。本发明提供的白背叶楤木挥发油可清除DPPH自由基和ABTS自由基，具有很好的抗氧化活性，可应用于制备抗氧剂，或用于制备食品、药品和/或化妆品，扩宽了白背叶楤木的应用领域，提升了其经济价值。
Description
技术领域
本发明属于天然产物领域，具体涉及一种白背叶楤木挥发油及其制备方法与应用。
背景技术
白背叶楤木（Aralia chinensis Linn. var. nuda Nakai），为五加科楤木属植物楤木（A. chinensis）的变种，生长于海拔1500 - 3000 m的森林或灌木丛中，为多年生落叶小乔木或灌木，俗称刺老苞椿头、刺头菜和刺老包，在云南西南地区分布较广。白背叶楤木药食两用，其嫩茎叶常作为野菜食用，气味清香，口感独特，可鲜食可烹饪。近年，楤木属植物因其较高营养及经济价值受到关注，研究发现，楤木属植物嫩芽富含多种氨基酸、维生素等营养物质，具有较高食用价值。龙牙楤木、太白楤木、辽东楤木等已实现人工栽培，楤木芽作为高级食材远销日本，被称为“天下第一山珍”。据《中国民族药志药》记载，白背叶楤木以根皮入药，可用于治疗跌打损伤，骨折，骨髓炎及深部脓疡，傈僳族民间医常将其单味使用，用于治疗风热咳喘、痢疾、黄疸、风湿痹痛等。现白背叶楤木已作为经济作物在云南省怒江傈僳族自治州泸水县进行推广种植，但目前对白背叶楤木挥发油成分的研究仍然空白。
发明内容
本发明的第一目的在于提供一种白背叶楤木挥发油，本发明的第二目的在于提供所述白背叶楤木挥发油的制备方法，本发明的第三目的是提供所述白背叶楤木挥发油的应用。
本发明的第一目的是这样实现的，一种白背叶楤木挥发油，以[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯并[E]薁-7-醇、植酮、棕榈酸甲酯、叶绿醇和α-松油醇为主要成分；按白背叶楤木挥发油的总重量计，[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯并[E]薁-7-醇的质量百分比含量为2～12%，植酮的质量百分比含量为0.2～3.5%，棕榈酸甲酯的质量百分比含量为0.2～2%，叶绿醇的质量百分比含量为0.2～17%，α-松油醇的质量百分比含量为0.1～1.5%。
本发明的第二目的是这样实现的，所述白背叶楤木挥发油的制备方法，按照以下步骤实现：
1）以白背叶楤木为原料，粉碎后加水浸泡4～10 h后置于提取器中用水蒸气蒸馏法于85～100℃下提取5～10 h，去掉水层，用有机溶剂反复洗涤提取器，得白背叶楤木挥发油粗提物；
2）所述白背叶楤木挥发油粗提物经无水硫酸钠干燥，过滤，滤液用旋转蒸发仪将有机溶剂挥干得到目标白背叶楤木挥发油。
本发明的第三目的是这样实现的，所述白背叶楤木挥发油的应用为在制备清除DPPH自由基和/或ABTS自由基的抗氧化活性药物或化妆品中的应用。
本发明具有如下有益效果：
1）本发明提供了一种白背叶楤木挥发油，所述挥发油中含有[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯并[E]薁-7-醇、植酮、棕榈酸甲酯、叶绿醇和α-松油醇等特征性成分，以白背叶楤木挥根部发油总量为100%计，所述挥发油中醇类成分含量不低于24.33%。本发明提供的白背叶楤木挥发油可清除DPPH自由基和ABTS自由基，具有很好的抗氧化活性，尤其是白背叶楤木根部挥发油，当质量浓度达到10.0 mg/mL时，挥发油对ABTS清除率与维生素C持平，可应用于制备抗氧剂，或用于制备食品、药品和/或化妆品，扩宽了白背叶楤木的应用领域，提升了其经济价值。
2）本发明制备白背叶楤木挥发油的方法简单，而且原料来源广，白背叶楤木的根、叶及茎都可以做为原料，适于工业化生产。
附图说明
图1为白背叶楤木根部挥发油总离子流图；
图2为白背叶楤木茎部挥发油总离子流图；
图3为白背叶楤木叶部挥发油总离子流图；
图4为白背叶楤木挥发油清除DPPH自由基能力曲线图；
图5为白背叶楤木挥发油清除ABTS自由基能力曲线图。
具体实施方式
下面结合附图与实施例对本发明作进一步的详细说明，但不以任何方式对本发明加以限制，基于本发明教导所作的任何变换或改进，均落入本发明的保护范围。
本发明一种白背叶楤木挥发油，以[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯并[E]薁-7-醇、植酮、棕榈酸甲酯、叶绿醇和α-松油醇为主要成分；按白背叶楤木挥发油的总重量计，[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯并[E]薁-7-醇的质量百分比含量为2～12%，植酮的质量百分比含量为0.2～3.5%，棕榈酸甲酯的质量百分比含量为0.2～2%，叶绿醇的质量百分比含量为0.2～17%，α-松油醇的质量百分比含量为0.1～1.5%。
本发明所述白背叶楤木挥发油的制备方法，按照以下步骤实现：
1）以白背叶楤木为原料，粉碎后加水浸泡4～10 h后置于提取器中用水蒸气蒸馏法于85～100℃下提取5～10 h，去掉水层，用有机溶剂反复洗涤提取器，得白背叶楤木挥发油粗提物；
2）所述白背叶楤木挥发油粗提物经无水硫酸钠干燥，过滤，滤液用旋转蒸发仪将有机溶剂挥干得到目标白背叶楤木挥发油。
步骤1中，浸泡时，原料与水的质量体积比为1:2～1:5 g/mL，浸泡时间为5～10 h。
水蒸气蒸馏法提取的温度为85～100℃。
步骤1中，将原料粉碎至20-80目。
步骤2中，有机溶剂为正己烷，旋转蒸发仪的温度为30℃～40℃。
原料为白背叶楤木的根、茎或叶的一种或几种的组合。
原料为根时，以白背叶楤木根部挥发油总量为100%计，白背叶楤木根部挥发油中醇类成分含量不低于24.33 %。
本发明所述白背叶楤木挥发油的应用为在制备清除DPPH自由基和/或ABTS自由基的抗氧化活性药物或化妆品中的应用。
本发明还提供了一种抗氧化剂，该抗氧化剂以所述白背叶楤木挥发油为唯一活性成分或活性成分之一。
以下结合实施例对本发明作进一步说明。
下述实施例中白背叶楤木采自云南省怒江傈僳族自治州泸水县鲁掌镇，由泸水县河新楤木种植农民专业合作社种植提供。由云南中医药大学中药学院李艳平副教授鉴定为白背叶楤木（Aralia chinensis Linn. var. nudaNakai ）的干燥根、茎、叶部位。
实施例中试剂及仪器信息如下：
正己烷（分析纯）：天津市风船化学试剂科技有限公司；无水硫酸钠（分析纯）：西陇科学股份有限公司； DPPH，ABTS（纯度≥98 %）：北京索莱宝科技有限公司；无水乙醇（分析纯）：天津市志远化学试剂有限公司；过硫酸钾（分析纯）：天津市津东天正精细化学试剂厂；抗坏血酸(VC)食品级：广东恒健制药有限公司。
气相色谱-质谱联用仪（GCMS-QP2010 Ultra）：日本岛津公司；挥发油提取器：四川蜀玻集团有限责任公司；调温电热套：北京中兴伟业仪器有限公司；高速粉碎机（DFY-C）：温岭市林大机械有限公司；多功能酶标仪（SparkTM 10M）：瑞士帝肯贸易有限公司。
实施例1 白背叶楤木根部挥发油的制备
取白背叶楤木干燥根300g，粉碎至40目，加入800 mL的水浸泡10 h，采用水蒸气蒸馏法于微沸状态下提取8h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得到白背叶楤木挥发油粗提物，在粗提物中加入无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在35℃回收正己烷得到4.3 g挥发油，得率为1.43%。
实施例2 白背叶楤木茎部挥发油的制备
取白背叶楤木茎300 g，粉碎至40目，加入800 mL的水浸泡10 h，采用水蒸气蒸馏法于微沸状态下提取8 h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并正己烷溶液，得白背叶楤木挥发油粗提物，加入粗无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在35℃下回收正己烷得到挥发油5.1 g，得率为1.70%。
实施例3 白背叶楤木叶部挥发油的制备
取白背叶楤木叶300 g，粉碎至40目，加入800 mL的水浸泡10 h，采用水蒸气蒸馏法于微沸状态下提取8 h时，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物；将得到的白背叶楤木挥发油粗提物用无水硫酸钠干燥，过滤，滤液用旋转蒸发仪于35℃下回收正己烷得到挥发油5.7 g，得率为1.90%。
实施例4 白背叶楤木根部挥发油的制备
取白背叶楤木根300 g，粉碎至20目，加入800 mL的水浸泡7 h，采用水蒸气蒸馏法于85℃下提取5h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物，在粗提物中加入无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在40℃回收正己烷得到4.0 g挥发油，得率为1.33%。
实施例5 白背叶楤木茎部挥发油的制备
取白背叶楤木茎300 g，粉碎至20目，加入800 mL的水浸泡7 h，采用水蒸气蒸馏法于90℃下提取5 h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物，加入粗无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在40℃下回收正己烷得到挥发油4.8 g，得率为1.60%。
实施例6 白背叶楤木叶部挥发油的制备
取白背叶楤木茎300g，粉碎至20目，加入800 mL的水浸泡7 h，采用水蒸气蒸馏法于92℃提取5 h时，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物；将得到的白背叶楤木挥发油粗提物用无水硫酸钠干燥，过滤，滤液用旋转蒸发仪于40℃下回收正己烷得到挥发油5.5g，得率为1.83%。
实施例7 白背叶楤木根部挥发油的制备
取白背叶楤木根300 g，粉碎至80目，加入1 L的水浸泡4 h，采用水蒸气蒸馏法于95℃提取10 h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物，在粗提物中加入无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在30℃回收正己烷得到4.5 g挥发油，得率为1.50 %。
实施例8 白背叶楤木茎部挥发油的制备
取白背叶楤木茎300g，粉碎至80目，加入1 L的水浸泡4 h，采用水蒸气蒸馏法于98℃下提取10 h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物，加入粗无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在30℃下回收正己烷得到挥发油5.4 g，得率为1.80%。
实施例9 白背叶楤木叶部挥发油的制备
取白背叶楤木茎300 g，粉碎至80目，加入1 L的水浸泡4 h，采用水蒸气蒸馏法于100℃下提取10 h，得到浮有挥发油的水溶液，将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得白背叶楤木挥发油粗提物；将得到的白背叶楤木挥发油粗提物用无水硫酸钠干燥，过滤，滤液用旋转蒸发仪于30℃下回收正己烷得到挥发油5.9g，得率为1.97%。
实验例1 白背叶楤木挥发油成分分析
对实施例1-3分别制备的根部挥发油、茎部挥发油和叶部挥发油的成分采用气相色谱-质谱联用仪分析。
气相色谱条件：色谱柱为SH-Rxi-5ms毛细管柱（30 m×0.25 mm×0.25 μm）毛细管柱，柱温45 ℃（保留2 min），以5 ℃/min升温至300 ℃，保持2 min，汽化室温度250 ℃，载气为高纯He，载气流速1.0 mL/min，进样量1 μL，分流比40:1，溶剂延迟时间3.0 min。
质谱条件：离子源温度230 ℃；四极杆温度150 ℃；接口温度280 ℃；电离方式为电子轰击离子源；电子能量70 eV；发射电流34.6 μA；质量数扫描范围m/z：20-450；SCAN全扫描模式。
按上述气相色谱和质谱条件对所得挥发油进行检查分析，由化学工作站绘出挥发油的总离子流图，总离子流图中的各峰经质谱扫描后得到质谱图，质谱图与Nist 2005和Wiley275标准质谱图集进行核对，结合数据库以及文献保留指数对比最终确定各化合物并按峰面积归一化法计算挥发油各组分的相对含量。
结果：实施例1-3分别制备的挥发油的总离子流图分别见图1-3,实施例1-3分别制备的挥发油的成分和相对含量结果分别见表1，从表1可知，实施例1制备的根部挥发油鉴定出化合物49种，醇类化合物占比最大，含量为24.33 %；实施例2制备的茎部挥发油鉴定出化合物39种，有机酸类物质占比最大，含量为33.82 %；实施例3制备的叶部挥发油鉴定出化合物73种，醇类占比最大，含量44.64 %。 从表1可知，根、茎和叶共有具有良好抗吸血寄生虫活性的高含量物质[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯[E]薁-7-醇，该物质首次在楤木属植物的挥发油中获得，且其含量在白背叶楤木的根部挥发油中含量高达11.44%，给[1aR-(1aα,4α,7β,7bα)]-1a,2,3,4,4a,5,6,7b-十氢-1,1,7-三甲基-4-亚甲基-1H-环丙烯[E]薁-7-醇的获得提供了一个新的途径。
表1 实施例1-3挥发油成分组成及含量分析
实验例2 白背叶楤木挥发油对DPPH自由基的清除能力测定
测定方法：分别取实施例1-3制备得到的挥发油，用无水乙醇配制成0.625 mg/mL、1.25 mg/mL、2.5 mg/mL、5 mg/mL、10 mg/mL的供试品溶液，取各浓度供试品溶液100 μL加入100 μL 2×10-4 mol/L DPPH溶液，室温反应30 min，于517 nm处测定吸光度A1，用等量无水乙醇代替DPPH溶液同法测定吸光度A2，以等量无水乙醇代替供试品溶液同法测定吸光度A0。以VC为阳性对照。各实验组平行3次，取平均值，清除率计算公式如下：
清除率（%）=（A0－A1+A2）/A0×100
结果：从图4可知，在质量浓度0-10 mg/mL范围内，实施例1-3制备得到的挥发油对DPPH自由基的清除能力均呈上升趋势，抗氧化能力：实施例1＞实施例3＞实施例2。其中实施例1根部挥发油清除DPPH自由基能力最强，当质量浓度达到10.0 mg/mL时，清除率达到84.0 %，IC50为4.97 mg/mL，表现出较强的抗氧化活性。
实验例3 白背叶楤木挥发油对ABTS自由基的清除能力测定
测定方法：分别取实施例1-3制备得到的挥发油，用无水乙醇配制成0.625 mg/mL、1.25 mg/mL、2.5 mg/mL、5 mg/mL、10 mg/mL的供试品溶液备用，用蒸馏水配制7 mmol/L的ABTS溶液和2.45 mol/L的过硫酸钾溶液，取过硫酸钾溶液88 μL加入5 mL ABTS溶液中，在室温黑暗处反应16 h，形成ABTS自由基储备液，在734 nm下用无水乙醇将ABTS自由基储备液稀释至吸光度0.7±0.02，备用。取配好的ABTS自由基储备液100 μL加入100 μL不同浓度供试品溶液室温反应30 min，于734 nm处测定吸光度A1，用等量无水乙醇代替ABTS自由基储备液同法测定吸光度A2，以等量无水乙醇代替供试品溶液同法测定吸光度A0。以VC为阳性对照。各实验组平行3次，取平均值，清除率计算公式如下：
清除率（%）=(A0－A1+A2)/A0×100
结果：从图5可以看出，实施例1-3制备得到的挥发油均具有一定的ABTS自由基的清除活性，其中实施例1制备得到的白背叶楤木根部挥发油对ABTS自由基清除能力最强，当挥发油质量浓度达到10.0 mg/mL时，挥发油对ABTS清除率与VC持平，IC50为2.90 mg/mL，说明当达到一定质量浓度时根部挥发油可表现较强的抗氧化活性。
Claims (3)
1.一种白背叶楤木挥发油，其特征在于，所述白背叶楤木挥发油按照以下步骤制备：
1）取白背叶楤木干燥根300g为原料粉碎至40目，加入800mL水浸泡10h，采用水蒸气蒸馏法于微沸状态下提取8h，得到浮有挥发油的水溶液；将水溶液的水层倒掉，用正己烷反复洗涤提取器，收集合并溶于正己烷溶液，得到白背叶楤木挥发油粗提物；
2）在粗提物中加入无水硫酸钠干燥，过滤，滤液用旋转蒸发仪在35℃回收正己烷得到4.3g挥发油，得率为1.43%。
2.一种权利要求1所述白背叶楤木挥发油在制备清除DPPH自由基和/或ABTS自由基的抗氧化活性药物或化妆品中的应用。
3.一种以权利要求1所述白背叶楤木挥发油为唯一活性成分或活性成分之一的抗氧化剂。
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| CN105670783A (zh) * | 2016-02-29 | 2016-06-15 | 广州市禾基生物科技有限公司 | 辽东楤木油及其提萃取方法 |
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2022
- 2022-06-22 CN CN202210712425.XA patent/CN115044407B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105670783A (zh) * | 2016-02-29 | 2016-06-15 | 广州市禾基生物科技有限公司 | 辽东楤木油及其提萃取方法 |
Non-Patent Citations (1)
| Title |
|---|
| 陈美航，等.棘茎楤木不同部位挥发油成分的比较.《中国实验方剂学杂志》.2013,第19卷(第12期),第124-128页. * |
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