雪松松针化学成分的研究
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摘要
雪松具有解痉、镇痛、抗炎、抗癌、抗菌、抗病毒等多种药理活性。其化学成分主要为非挥发性成分和挥发性成分两部分,其中非挥发性成分主要含萜类、木脂素类、黄酮类、脂肪酸等;挥发性成分中以萜类化合物为多见,主要是单萜、倍半萜和它们的含氧衍生物。本论文归纳总结了国内外学者对雪松化学成分及药理活性的研究成果,希冀为雪松的全面开发利用和深入研究提供参考。
本论文对雪松松针中挥发油进行成分分析研究,并建立了非挥发性成分总黄酮的含量测定方法,优选了提取工艺路线。
为研究雪松松针中挥发油的化学成分,将雪松松针用常规水蒸汽蒸馏制得挥发油,出油率为0.1%;采用气相色谱/质谱法(Gas chromatography/massspectrometry method)对其挥发油的化学成分进行分析,结果分离得到83个峰,鉴定出68个组分,占总挥发油的96.56%,主要化学成分为2,5-三亚甲基降茨烷、(+)-(1S,3S)-间薄荷-4,8-二烯、D-柠檬烯、a-松油醇、4-萜烯醇、石竹烯、石竹烯氧化物等,其中2,5-三亚甲基降茨烷(32.65%)、(+)-(1S,3S)-间薄荷-4,8-二烯(21.45%)、D-柠檬烯(13.72%)、a-松油醇(5.75%)、4-萜烯醇(1.46%)、石竹烯(1.41%)、石竹烯氧化物(1.25%)、1-甲基-4-异丙基苯(1.10%),占总挥发油的78.79%,此外还含有酸、酯、烯、醇、含氮氧化物等多种化合物。雪松松针中的黄酮类化合物为重要的生理活性物质,本文根据黄酮类化合物与金属Al~(3+)形成的螯合物在410nm波长处有最大吸收的原理,用无水乙醇提取雪松松针,以芦丁为对照品,建立雪松松针中总黄酮含量测定的紫外分光光度法(Ultravioletspectrophotometry),结果显示芦丁在4.46～26.76μg·ml~(-1)范围内线性关系良好,回归方程为A=27.338C-0.0086,r=0.9991(n=3);平均回收率为100.4%,RSD=0.95%,该方法简便、准确、可靠,可作为雪松松针中总黄酮的含量检测方法。为优选雪松松针中总黄酮的最佳提取工艺,以总黄酮的含量为检测指标,采用L_9(3~4)正交试验对雪松松针的超声提取工艺进行研究。正交试验结果表明:影响提取工艺的因素依次为温度>提取时间>固液比,最终确定的雪松松针中总黄酮的提取工艺为:用30倍的无水乙醇,于40℃下超声提取3次,每次20min。经验证,该工艺简便快速,合理可行。
The major pharmacological activities of genus Cedrus deodara include analgesic, spasm, anti-inflammatory, anticancer, antibacterial, anti-virus etc. And mainly chemical constituents include two parts: non-volatile and volatile constituents.Non-volatile constituents include mainly terpenes, lignans, flavonoids, fatty acids etc.the main volatile components are terpenoids components, including monoterpenes,sesquiterpenes and their oxygenated derivatives.This paper summarizes research current status of genus Cedrus deodara in domestic and foreign , provide the reference on comprehensive development and utilization of genus Cedrus deodara.
In this paper, constituents of volatile oil from Pine needles of Cedrus deodara were studied and the extraction technics routes of total flavonoids from Pine needles were optimized.
The volatile oil of Pine needles was obtained by steam distillation, The result shown that the yield of volatile oil from Pine needles is 0.1%, and its chemical compounds were analyzed by Gas Chromatography/mass spectrometry, 83 peaks were isolated and 68 compounds were identified,which accounted for 96.56% of the total contents,the main chemical compounds are 2,5-Trimethylenenorbornane (32.65%), m-Mentha-4,8-diene(21.45%),D-limonen(13.72%),p-Menth-l-en-8-ol(5.75%),4-Terpineol(1.46%),caryophyllene(1.41%),caryophyllene oxide(1.25%),Benzene,l-methyl-4-(l-methylethyl)(1.10%),which accounted for 78.79% of the total contents,including acid,ester,hydroxybenzene,nitrogenoxide and so on.The total flavonoids is an important pharmacological activities.In this context Pine needles were extracted by ethanol ,rutin was used as reference substance, detection wavelength was 410nm,the method was established to determine total flavonoids content of pine needles by ultraviolet spectrophotometry.The result shown that the linearity of rutin was obtained within the range of 4.46～26.76μg·ml~(-1),Regression Equation Was A=27.338C-0.0086, and correlation coefficient r=0.9991(n=3).The average recovery was 100.4% and RSD =0.95%.The method is simple, rapid and accurate.It can be used as a quality control method to determine the total flavonoids of Pine needles.To optimize the best extraction technics routes of total flavonoids from Pine needles, that total flavornoids was used as detective target, the best ultrasonic wave extraction technics routes was studied by orthogonal design of L_9 (3~4) .The result was indicated that the order of factors influenced to affect extraction was temperature > extraction time > liquid solid ratio.The optimum extraction technics conditions were 30-fold ethanol, extracting by ultrasonic wave for 20minutes each at 40℃for three times.The best extraction technic routes reasonable, practicable and convenient.
本论文对雪松松针中挥发油进行成分分析研究,并建立了非挥发性成分总黄酮的含量测定方法,优选了提取工艺路线。
为研究雪松松针中挥发油的化学成分,将雪松松针用常规水蒸汽蒸馏制得挥发油,出油率为0.1%;采用气相色谱/质谱法(Gas chromatography/massspectrometry method)对其挥发油的化学成分进行分析,结果分离得到83个峰,鉴定出68个组分,占总挥发油的96.56%,主要化学成分为2,5-三亚甲基降茨烷、(+)-(1S,3S)-间薄荷-4,8-二烯、D-柠檬烯、a-松油醇、4-萜烯醇、石竹烯、石竹烯氧化物等,其中2,5-三亚甲基降茨烷(32.65%)、(+)-(1S,3S)-间薄荷-4,8-二烯(21.45%)、D-柠檬烯(13.72%)、a-松油醇(5.75%)、4-萜烯醇(1.46%)、石竹烯(1.41%)、石竹烯氧化物(1.25%)、1-甲基-4-异丙基苯(1.10%),占总挥发油的78.79%,此外还含有酸、酯、烯、醇、含氮氧化物等多种化合物。雪松松针中的黄酮类化合物为重要的生理活性物质,本文根据黄酮类化合物与金属Al~(3+)形成的螯合物在410nm波长处有最大吸收的原理,用无水乙醇提取雪松松针,以芦丁为对照品,建立雪松松针中总黄酮含量测定的紫外分光光度法(Ultravioletspectrophotometry),结果显示芦丁在4.46～26.76μg·ml~(-1)范围内线性关系良好,回归方程为A=27.338C-0.0086,r=0.9991(n=3);平均回收率为100.4%,RSD=0.95%,该方法简便、准确、可靠,可作为雪松松针中总黄酮的含量检测方法。为优选雪松松针中总黄酮的最佳提取工艺,以总黄酮的含量为检测指标,采用L_9(3~4)正交试验对雪松松针的超声提取工艺进行研究。正交试验结果表明:影响提取工艺的因素依次为温度>提取时间>固液比,最终确定的雪松松针中总黄酮的提取工艺为:用30倍的无水乙醇,于40℃下超声提取3次,每次20min。经验证,该工艺简便快速,合理可行。
The major pharmacological activities of genus Cedrus deodara include analgesic, spasm, anti-inflammatory, anticancer, antibacterial, anti-virus etc. And mainly chemical constituents include two parts: non-volatile and volatile constituents.Non-volatile constituents include mainly terpenes, lignans, flavonoids, fatty acids etc.the main volatile components are terpenoids components, including monoterpenes,sesquiterpenes and their oxygenated derivatives.This paper summarizes research current status of genus Cedrus deodara in domestic and foreign , provide the reference on comprehensive development and utilization of genus Cedrus deodara.
In this paper, constituents of volatile oil from Pine needles of Cedrus deodara were studied and the extraction technics routes of total flavonoids from Pine needles were optimized.
The volatile oil of Pine needles was obtained by steam distillation, The result shown that the yield of volatile oil from Pine needles is 0.1%, and its chemical compounds were analyzed by Gas Chromatography/mass spectrometry, 83 peaks were isolated and 68 compounds were identified,which accounted for 96.56% of the total contents,the main chemical compounds are 2,5-Trimethylenenorbornane (32.65%), m-Mentha-4,8-diene(21.45%),D-limonen(13.72%),p-Menth-l-en-8-ol(5.75%),4-Terpineol(1.46%),caryophyllene(1.41%),caryophyllene oxide(1.25%),Benzene,l-methyl-4-(l-methylethyl)(1.10%),which accounted for 78.79% of the total contents,including acid,ester,hydroxybenzene,nitrogenoxide and so on.The total flavonoids is an important pharmacological activities.In this context Pine needles were extracted by ethanol ,rutin was used as reference substance, detection wavelength was 410nm,the method was established to determine total flavonoids content of pine needles by ultraviolet spectrophotometry.The result shown that the linearity of rutin was obtained within the range of 4.46～26.76μg·ml~(-1),Regression Equation Was A=27.338C-0.0086, and correlation coefficient r=0.9991(n=3).The average recovery was 100.4% and RSD =0.95%.The method is simple, rapid and accurate.It can be used as a quality control method to determine the total flavonoids of Pine needles.To optimize the best extraction technics routes of total flavonoids from Pine needles, that total flavornoids was used as detective target, the best ultrasonic wave extraction technics routes was studied by orthogonal design of L_9 (3~4) .The result was indicated that the order of factors influenced to affect extraction was temperature > extraction time > liquid solid ratio.The optimum extraction technics conditions were 30-fold ethanol, extracting by ultrasonic wave for 20minutes each at 40℃for three times.The best extraction technic routes reasonable, practicable and convenient.
引文
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[2]Miller,CN.Mesozoicconifers[J].BotanicalReview,1997,43(2):217-218.
[3]李斌,顾万春,卢宝铭.白皮松天然群体种实性状表型多样性研究[J].生物多样性,2002,10(2):181-187.
[4]中草药学编写组.中草药学.南京:江苏人民出版社,1978.
[5]杨天明,夏德超,朱景申.松针的研究进展[J]中国药师,2002,5(12):748-749.
[6]李萍,刘友平.松针研究进展[J]成都中医药大学学报,2001,24(3):49-50.
[7]毕跃峰,郑晓珂,冯卫生,等.[J]松针的研究进展及开发利用,199,14(2):14-17.
[8]郑万钧,傅立国.中国植物志(第七卷)[M].北京:科学出版社,1978:167-203.
[9]Dallimore W, Jac kson A B, Harrion S GHandb Conif and Ginkgo, ed.4 [M].London:Edward, 1966:137-146.
[10]Mhaskar K S, Blatter E, Caius (Eds.) J F. Indian Medicinal Plants,vol.3[M].New Delhi:Satguru Publications,2000:3303.
[11]Sachin B S,Koul M,Zuthshi A,et al. Simultaneous high-performance liquid Chromatographic determination of Cedrus deodara active constituents and the pharmacokinetic profile in mice [J].J Chromatogr B,2008,862:237-241.
[12]Joseph T C, Sukh D. Studies in sesquiterpenes-ⅩⅩⅨ structure of himachalenes [J].Tetrahedron,1968,24:3809-3827.
[13]Joseph T C,Sukh D.Parts ⅩⅩⅨ-ⅩⅩⅩⅡ of the present series:studies in sesquiterpenes[J].Tetrahedron,1961,17:216.
[14]Bisarya S C, Sukh D.Studies in sesquiterpenes-ⅩⅩⅩⅢ: himachalol[J].Tetrahedron, 1968,24:3861-3867.
[15]Bisarya S C,Sukh D.Studies in sesquiterpenes-:ⅩⅩⅩⅣ:structure of allohimachalol [J].Tetrahedron,1968,24:3869-3879.
[16]Kulshreshtha D K, Rastogi R P.Constitution of the sesquiterpenod, centdarol[J].Phytochemistry,1975,14(10):2237-2240.
[17]Kulshreshtha D K, Rastogi R P.The constitution of isocentdarol,a sesquiterpenediol from Cedrus deodara[J].hytochemistry,1976,15(4):557-558.
[18]Pande B S,Krishnappa S,Bisarya S C,et al.Studies in sesquiterpenes -XLVII: Cis-and trans-atlantones from Cedrus deodara loud[J].Tetrahedron,1971,27(4):841-844.
[19]Shinde U A, Phadke A S,Nair A M, et al.Membrane stabilizing activityapossible mechanism of action forthe anti-inflammatoryactivity of Cedrus deodara wood oil [J].Fitoterapia,1999,70:251-257.
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