GC/MS法对龙脑樟的质量评价研究
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摘要
目的：
龙脑樟Cinnamomum camphora (L.) Presl为亚热带常绿阔叶乔木,其枝叶是提取天然冰片的最佳植物资源。是属于樟树中的一个稳定的化学类型—-龙脑型樟树。
龙脑樟中所含的天然右旋龙脑(天然D-龙脑),又称天然冰片,为中医常用的著名芳香开窍中药,是安宫牛黄丸、丹参滴丸、六神丸等中成药的主要组分。过去,天然冰片是采用龙脑香科龙脑香(Dryobalanops aroma tica GaerthnF.)树的树脂提取,这种植物主要分布在印尼的苏门答腊群岛,国内几乎没有,因此过去主要依靠进口,龙脑樟树的发现及开发改变了这一状况。不过目前天然冰片的供应仍紧张。为了使天然冰片产量增高,对天然冰片的最佳植物来源—龙脑樟进行质量评价研究越来越重要。
本实验目的是采用GC／MS法考察不同采收期、不同产地、不同部位对龙脑樟中D-龙脑含量的影响,为龙脑樟的质量评价提供依据；对不同部位龙脑樟挥发油进行成分分析,有利于综合开发与利用龙脑樟中的天然D-龙脑以及挥发油中其他活性成分。
方法：
1.龙脑樟叶中天然D-龙脑最佳提取工艺研究：以水蒸气蒸馏法提取,应用正交试验设计选取D-龙脑最佳提取工艺条件,用GC／MS法进行含量测定,以龙脑樟中D-龙脑含量为指标,考察料液比、浸泡时间、提取时间三因素,探讨D-龙脑的最佳提取工艺。
2.龙脑樟叶最佳采收期研究：采收不同生育期的龙脑樟叶,用内标法测定D-龙脑含量,研究龙脑樟叶的D-龙脑的含量动态分布变化规律,从而确定最佳采收期。
3.不同产地、不同部位龙脑樟的D-龙脑的含量测定：用内标法测定D-龙脑含量,比较研究龙脑樟(8月份采收)不同部位的D-龙脑的含量。
4.龙脑樟植物不同部位的挥发油成分分析：采用GC／MS法对龙脑樟的不同部位挥发油进行成分分析,并比较研究各部位挥发油化学组成以及挥发油中各组分的含量异同。
结果：
1.以D-龙脑含量为指标,龙脑樟最佳提取工艺为：水蒸气蒸馏法进行提取,料液比1：20,浸泡0h,提取2h。
该法在0.4～2.8mg／ml浓度范围内线性良好,R2=0.9992。
2.龙脑樟叶最佳采收期研究：龙脑樟叶中D-龙脑的含量以8月份含量最高,2月份含量最低。
3.不同产地龙脑樟叶的D-龙脑的含量测定：新晃龙脑樟叶>浏阳龙脑樟叶>长沙县龙脑樟叶。
4.不同部位龙脑樟的D-龙脑的含量测定：老叶>果实>新叶>枝心>枝皮。
5.不同部位龙脑樟挥发油的成分分析结果：
从龙脑樟5种挥发油中共鉴定出77种成分,从老叶、新叶、枝皮、枝心及果实分别鉴定出了52、48、55、28和49种化合物,已鉴定的组分分别占相应挥发油的99.40%、99.49%、97.12%、88.86%和99.42%。
龙脑樟各个不同部位(老叶,新叶,枝皮,枝心,果实)中挥发油的共有成分有19种,分别为D-龙脑,樟脑,乙酸正丁酯,α-侧柏烯,α-蒎烯,莰烯,β-蒎烯,β-月桂烯,α-水芹烯,α-伞花烃,D-柠檬烯,桉叶油素,Y-萜烯,顺-β-罗勒烯,β-芳樟醇,α-松油醇,β-石竹烯,α-石竹烯,石竹烯氧化物。
结论：
1.建立了龙脑樟中D-龙脑的GC／MS含量测定法,该GC/MS法准确、快速、稳定,重现性好。该法在0.4～2.8mg／ml浓度范围内线性良好,R2=0.9992。
龙脑樟最佳提取工艺为：水蒸气蒸馏法进行提取,料液比1：20,提取前不需浸泡,提取时间为2个小时。
2.龙脑樟叶的D-龙脑含量在8月份最高,且8月份樟叶处于盛叶期,因此最佳采收时期为8月份。
3.龙脑樟各个部位中,以老叶D-龙脑含量最高,果实次之；三个产地中,以新晃县的龙脑樟叶D-含量最高。
4.龙脑樟不同部位的挥发性成分相差较大,在综合开发利用龙脑樟挥发油时应分部位(老叶,新叶,枝皮,枝心,果实)加以提取利用。
Objective：Cinnamomum camphora (L.) Presl is a kind of evergreen broadleaved tree, and its branch leaves are the optimal plant resource of natural borneol production. It is one of the chemical types of Cinnomomum camphora (L.) Presl, which is stable-----borneol type of Cinnomomum camphora (L.) Presl.
Natural D-borneol contained in Cinnamomum camphora (L.) Presl is also called as natural borneol, which is a famous and frequently-used Chinese drug for inducing resuscitation, and is the main component of Chinese traditional medicine, such as An gong niu huang Bolus, Dan shen Bolus and Liu shen Pill and so on.To increase the production of natural borneol, it's more and more important to study on Cinnomomum camphora (L.) Presl.
The main objective of the article is to utilize D-borneol and other active components in violate oil of Cinnomomum camphora (L.) Presl more comprehensively, which is beneficial to utilize violate oil of Cinnomomum camphora (L.) Presl comprehensively.
Method:
1. Study on extraction technology of D-borneol from Cinnamomum camphora (L.) Presl:The optimum extraction condition was investigated by the orthogonal design L9(34). The orthogonal test method was used for studying the effects of three factors(solid-liquid ratio,the distillating time, and the soaking time).
2. The study of the Seasonal Variations of the Contents of D-borneol in leaves of Cinnamomum camphor a (L.) Presl:Using internal standard method to determine the content of D-borneol of leaves collected in differrent harvest time, to study the dynamic change law of content of D-borneol.
3. Study on D-borneol content measuration of different parts of Cinnamomum camphora (L.) Presl:Using internal standard method to determine the content of D-borneol of different parts (August), to compare and analyze the similarities and differences.
4. Analysis of chemical constituents of volatile oil from different parts(old leaf, young leaf, branch cortex, branch core, fruit) of Cinnamomum camphora (L.) Presl in the same collecting season.
Results:
1. The optimum extraction conditions of D-borneol are:solid-liquid ratio 1:20, soaking time 0 h, and distillating time 2h.
The linearity and correlation coefficient was yielded from 0.4-2.8mg/ml (R2=0.9994).
2. The study of the Seasonal Variations of the Contents of D-borneol in leaves of Cinnamomum camphora (L.) Presl:The content of D-borneol determined is highest in August, lowest in December.
3.Study on D-borneol content measuration of different parts of Cinnamomum camphora (L.) Presl:The content of D-borneol determined is highest in leaf, lowest in branch cortex.
4. Constituent analysis of volatile oil among different parts of Cinnamomum camphora (L.) Presl in the same collecting season:
The total number of chemical contituents of the five kinds of volatile oil are 77, and the each number of chemical contituents of five kinds of volatile oil is 52、48、55、28、51, the content of the contituents confirmed is:99.40%,99.49%,97.12%, 88.86%和99.42%.
The common constituents of volatile oil from five different parts are 19 constituents:D-Borneol, comphor, n-butyl acetate, a-Thujene, a-Pinene, Camphene,β-Pinene, p-Myrcene, a-Phellandrene, o-Cymene, D-Limonene, Cineole, cis-β-Ocimene, y-Terpinene, cis-p-Ocimene, P-Linalool, a-Terpineol, P-Caryophyllene, a-Caryophyllene, Caryophyllene oxide.
Conclusions:
1.It had established GC/MS method to determine the content of D-borneol in Cinnamomum camphora (L.) Presl, GC/MS method is accurate and reliable.
The linearity and correlation coefficient was yielded from 0.4-2.8mg/ml (R2=0.9992)
The optimum extraction conditions of D-borneol are:solid-liquid ratio 1:20, soaking time 0 h, and distillating time 2h.
2.The highest content period of D-borneol in leaves of Cinnamomum camphora (L.) Presl was August, so August is the best time to collect the leaves.
3.Among the five parts, leaves is the best part to extract D-borneol, fruit is the second.
4.The chemical constituents of each part of Cinnamomum camphora (L.) Presl are different from others, so we can extract violate oil from each part, to utilize violate oil of Cinnomomum camphora (L.) Presl comprehensively.
龙脑樟Cinnamomum camphora (L.) Presl为亚热带常绿阔叶乔木,其枝叶是提取天然冰片的最佳植物资源。是属于樟树中的一个稳定的化学类型—-龙脑型樟树。
龙脑樟中所含的天然右旋龙脑(天然D-龙脑),又称天然冰片,为中医常用的著名芳香开窍中药,是安宫牛黄丸、丹参滴丸、六神丸等中成药的主要组分。过去,天然冰片是采用龙脑香科龙脑香(Dryobalanops aroma tica GaerthnF.)树的树脂提取,这种植物主要分布在印尼的苏门答腊群岛,国内几乎没有,因此过去主要依靠进口,龙脑樟树的发现及开发改变了这一状况。不过目前天然冰片的供应仍紧张。为了使天然冰片产量增高,对天然冰片的最佳植物来源—龙脑樟进行质量评价研究越来越重要。
本实验目的是采用GC／MS法考察不同采收期、不同产地、不同部位对龙脑樟中D-龙脑含量的影响,为龙脑樟的质量评价提供依据；对不同部位龙脑樟挥发油进行成分分析,有利于综合开发与利用龙脑樟中的天然D-龙脑以及挥发油中其他活性成分。
方法：
1.龙脑樟叶中天然D-龙脑最佳提取工艺研究：以水蒸气蒸馏法提取,应用正交试验设计选取D-龙脑最佳提取工艺条件,用GC／MS法进行含量测定,以龙脑樟中D-龙脑含量为指标,考察料液比、浸泡时间、提取时间三因素,探讨D-龙脑的最佳提取工艺。
2.龙脑樟叶最佳采收期研究：采收不同生育期的龙脑樟叶,用内标法测定D-龙脑含量,研究龙脑樟叶的D-龙脑的含量动态分布变化规律,从而确定最佳采收期。
3.不同产地、不同部位龙脑樟的D-龙脑的含量测定：用内标法测定D-龙脑含量,比较研究龙脑樟(8月份采收)不同部位的D-龙脑的含量。
4.龙脑樟植物不同部位的挥发油成分分析：采用GC／MS法对龙脑樟的不同部位挥发油进行成分分析,并比较研究各部位挥发油化学组成以及挥发油中各组分的含量异同。
结果：
1.以D-龙脑含量为指标,龙脑樟最佳提取工艺为：水蒸气蒸馏法进行提取,料液比1：20,浸泡0h,提取2h。
该法在0.4～2.8mg／ml浓度范围内线性良好,R2=0.9992。
2.龙脑樟叶最佳采收期研究：龙脑樟叶中D-龙脑的含量以8月份含量最高,2月份含量最低。
3.不同产地龙脑樟叶的D-龙脑的含量测定：新晃龙脑樟叶>浏阳龙脑樟叶>长沙县龙脑樟叶。
4.不同部位龙脑樟的D-龙脑的含量测定：老叶>果实>新叶>枝心>枝皮。
5.不同部位龙脑樟挥发油的成分分析结果：
从龙脑樟5种挥发油中共鉴定出77种成分,从老叶、新叶、枝皮、枝心及果实分别鉴定出了52、48、55、28和49种化合物,已鉴定的组分分别占相应挥发油的99.40%、99.49%、97.12%、88.86%和99.42%。
龙脑樟各个不同部位(老叶,新叶,枝皮,枝心,果实)中挥发油的共有成分有19种,分别为D-龙脑,樟脑,乙酸正丁酯,α-侧柏烯,α-蒎烯,莰烯,β-蒎烯,β-月桂烯,α-水芹烯,α-伞花烃,D-柠檬烯,桉叶油素,Y-萜烯,顺-β-罗勒烯,β-芳樟醇,α-松油醇,β-石竹烯,α-石竹烯,石竹烯氧化物。
结论：
1.建立了龙脑樟中D-龙脑的GC／MS含量测定法,该GC/MS法准确、快速、稳定,重现性好。该法在0.4～2.8mg／ml浓度范围内线性良好,R2=0.9992。
龙脑樟最佳提取工艺为：水蒸气蒸馏法进行提取,料液比1：20,提取前不需浸泡,提取时间为2个小时。
2.龙脑樟叶的D-龙脑含量在8月份最高,且8月份樟叶处于盛叶期,因此最佳采收时期为8月份。
3.龙脑樟各个部位中,以老叶D-龙脑含量最高,果实次之；三个产地中,以新晃县的龙脑樟叶D-含量最高。
4.龙脑樟不同部位的挥发性成分相差较大,在综合开发利用龙脑樟挥发油时应分部位(老叶,新叶,枝皮,枝心,果实)加以提取利用。
Objective：Cinnamomum camphora (L.) Presl is a kind of evergreen broadleaved tree, and its branch leaves are the optimal plant resource of natural borneol production. It is one of the chemical types of Cinnomomum camphora (L.) Presl, which is stable-----borneol type of Cinnomomum camphora (L.) Presl.
Natural D-borneol contained in Cinnamomum camphora (L.) Presl is also called as natural borneol, which is a famous and frequently-used Chinese drug for inducing resuscitation, and is the main component of Chinese traditional medicine, such as An gong niu huang Bolus, Dan shen Bolus and Liu shen Pill and so on.To increase the production of natural borneol, it's more and more important to study on Cinnomomum camphora (L.) Presl.
The main objective of the article is to utilize D-borneol and other active components in violate oil of Cinnomomum camphora (L.) Presl more comprehensively, which is beneficial to utilize violate oil of Cinnomomum camphora (L.) Presl comprehensively.
Method:
1. Study on extraction technology of D-borneol from Cinnamomum camphora (L.) Presl:The optimum extraction condition was investigated by the orthogonal design L9(34). The orthogonal test method was used for studying the effects of three factors(solid-liquid ratio,the distillating time, and the soaking time).
2. The study of the Seasonal Variations of the Contents of D-borneol in leaves of Cinnamomum camphor a (L.) Presl:Using internal standard method to determine the content of D-borneol of leaves collected in differrent harvest time, to study the dynamic change law of content of D-borneol.
3. Study on D-borneol content measuration of different parts of Cinnamomum camphora (L.) Presl:Using internal standard method to determine the content of D-borneol of different parts (August), to compare and analyze the similarities and differences.
4. Analysis of chemical constituents of volatile oil from different parts(old leaf, young leaf, branch cortex, branch core, fruit) of Cinnamomum camphora (L.) Presl in the same collecting season.
Results:
1. The optimum extraction conditions of D-borneol are:solid-liquid ratio 1:20, soaking time 0 h, and distillating time 2h.
The linearity and correlation coefficient was yielded from 0.4-2.8mg/ml (R2=0.9994).
2. The study of the Seasonal Variations of the Contents of D-borneol in leaves of Cinnamomum camphora (L.) Presl:The content of D-borneol determined is highest in August, lowest in December.
3.Study on D-borneol content measuration of different parts of Cinnamomum camphora (L.) Presl:The content of D-borneol determined is highest in leaf, lowest in branch cortex.
4. Constituent analysis of volatile oil among different parts of Cinnamomum camphora (L.) Presl in the same collecting season:
The total number of chemical contituents of the five kinds of volatile oil are 77, and the each number of chemical contituents of five kinds of volatile oil is 52、48、55、28、51, the content of the contituents confirmed is:99.40%,99.49%,97.12%, 88.86%和99.42%.
The common constituents of volatile oil from five different parts are 19 constituents:D-Borneol, comphor, n-butyl acetate, a-Thujene, a-Pinene, Camphene,β-Pinene, p-Myrcene, a-Phellandrene, o-Cymene, D-Limonene, Cineole, cis-β-Ocimene, y-Terpinene, cis-p-Ocimene, P-Linalool, a-Terpineol, P-Caryophyllene, a-Caryophyllene, Caryophyllene oxide.
Conclusions:
1.It had established GC/MS method to determine the content of D-borneol in Cinnamomum camphora (L.) Presl, GC/MS method is accurate and reliable.
The linearity and correlation coefficient was yielded from 0.4-2.8mg/ml (R2=0.9992)
The optimum extraction conditions of D-borneol are:solid-liquid ratio 1:20, soaking time 0 h, and distillating time 2h.
2.The highest content period of D-borneol in leaves of Cinnamomum camphora (L.) Presl was August, so August is the best time to collect the leaves.
3.Among the five parts, leaves is the best part to extract D-borneol, fruit is the second.
4.The chemical constituents of each part of Cinnamomum camphora (L.) Presl are different from others, so we can extract violate oil from each part, to utilize violate oil of Cinnomomum camphora (L.) Presl comprehensively.
引文
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[21]陈铁峰,林曙光,陈露西,等.冰片促进四甲基吡嗪的吸收[J].中国药理学报,1990,11(1)：42-44.
[22]杨丽莉,袁倚胜,佟永岭,等.冰片和川芎嗪血药浓度的c,C-MSD测定法[J].药学学报,1994,29(9)：697-701.
[23]王原,吴伟康,黄志宏,等.冰片在正常人皮肤增加曲安缩松的透皮生物利用度[J].中药药理与临床,1992,8(特辑)：178.
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[1]王坚.樟科樟属植物樟的化学成分和组织培养研究[J].江西中医学院学报,2004,16(2)：69—70.
[2]周汉芬,韦一良,胡健华,等.樟树籽仁油醇解工艺研究[J].中国油脂,2005,30(8)： 53-56.
[3]欧阳少林,龙光远,黄璐琦,等.天然冰片的新资源[J].江西林业科技.2005,5：.38.
[4]龙光远,彭招兰,郭德选,等.龙脑樟矮林作业技术和效益分析[J].林业科技开发,2000,14(6)：30-31.
[5]程平,蓝裕民,黄宝祥,等.樟树籽核油的开发利用[J].江西林业科技,1998(1)： 26-27.
[6]石皖阳,何伟,文光裕,等.樟精油成分和类型划分[J].植物学报,1989,31(3)： 209-214.
[7]陈金芳,张良均,童身毅.樟树籽及其综合利用[J].林产化工通讯,1991：24-26.
[8]周武,张彬,邓丹雯.樟树籽油提取的研究[J].中国油脂,2004,29(2)：30一31.
[9]李林松,罗永明.井岗山地区樟树果挥发性成分的分析[J].江西中医学院学报.2005,6(17)：36-37.
[10]刘亚,李茂昌,张承聪,等.香樟树叶挥发油的化学成分研究[J].分析实 验室.2008,1(27)：88-92.
[11]张斌,许莉勇.樟树籽提取樟油的试验[J].浙江林学院学报2001,18(1)：57-59.
[12]戴宝合.野生植物资源学[R].北京：农业出版社,1993
[13]陈美兰,黄璐琦,等.药用植物樟化学型形成机理的基础研究[R].中国中医科学院：17-18.
[14]Prates H T, Santos J P, Waquil J M, et al. Ovicidal activity of monoterpenes against RhyzoPertha dominica (F.) and Tribolium castaneum (Herbst) [J]. J. Stored Prod. Res.1998,34 (4):243-249.
[15]Cimanga K, Kambu, K, Tona L, et. al. Correlation between chemical composition and antibacterial activity of essential oils of some aromatic medicinal Plants growing In the Democratic Republic of Congo [J]. Journal of EthnoPharmacology,2002,79(2):213-220.
[16]田玉红,张祥民,等.桉叶油的研究进展.食品与发酵工业[J].2007(33)：139-143.
[17]吴桂生.冰片中毒的救治方法.中华腹部疾病杂志[J],2006,6(7)：517-518.
[18]陈建南.曾惠芳,李耿,等.龙脑樟挥发油及天然冰片成分分析[J].中药材,2005,28(9)：782—783.
[19]魏雪芳,陈杰.冰片提高生物利用度的研究进展[J].中草药,2005,36(7)：1106—1108.
[20]朱健平,王宗锐,吴宋夏.龙脑促进药物经皮渗透作用的研究[J].中国药学杂志,1999,34(2)：104.
[21]陈铁峰,林曙光,陈露西,等.冰片促进四甲基吡嗪的吸收[J].中国药理学报,1990,11(1)：42-44.
[22]杨丽莉,袁倚胜,佟永岭,等.冰片和川芎嗪血药浓度的c,C-MSD测定法[J].药学学报,1994,29(9)：697-701.
[23]王原,吴伟康,黄志宏,等.冰片在正常人皮肤增加曲安缩松的透皮生物利用度[J].中药药理与临床,1992,8(特辑)：178.
[24]王晖,许卫铭,王宗锐,等.薄荷醇与冰片的促透作用[J].中草药,1997.28(2)：93-95.
[25]朱健平,王宗锐,吴宋夏.龙脑促进药物经皮渗透作用的研究[J].中国药学杂志,1999,34(2)：104-106.
[26]许碧莲,王晖,许卫铭.冰片对盐酸川芎嗪促透作用的研究[J].中成药,2001,23(12)：864—867.
[27]江文德,徐端正,胡国钧,等.冠心苏合丸的药理研究及其简化制剂-苏冰滴丸的理论基础[J].药学学报,1979,14(11)：655-660.
[28]邓文龙.开窍方药的临床及实验研究进展[J].中成药研究,1985,(3)：33-38.
[29]江光池,冯旭军,黄岚,等.龙脑和异龙脑对小鼠和家兔的药理作用[J].华西药学杂志,1989,4(1)：23-25.
[30]刘德福,刘强,孙以林,等.麝香和冰片促进雪旺细胞生长作用的研究[J].哈尔滨医科大学学报,1986,(4)：6-7.
[31]牟家琬,杨胜华,孙玉梅,等.龙脑和异龙脑的体外抗菌作用的研究[J].华西药学杂志,1989,4(1)：20-22.
[32]江光池,杨胜华,冯旭军.龙脑和异龙脑的抗炎作用[J].华西药学杂志,1990,5(3)：190-191.
[33]侯桂芝,廖仁德,孟如松,等.冰片对激光烧伤创面的镇痛及抗炎作用[J].中国药学杂志,1995,30(9)：532-534.
[34]徐连英,陶建生,冯怡.冰片抗生育作用及剂型研究[J].中成药研究,1986,(3)：1.