华中五味子果实发育阶段木脂素含量变化的研究
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摘要
华中五味子(Schisandra sphenanthera Rehd.et Wils.)为木兰科五味子属的落叶藤本,其干燥果实被称为南五味子。由于其含量丰富的木脂素类物质具有许多生物学活性,一直以来,作为一种传统中药,被广泛运用于中医治疗。我国华中五味子野生资源丰富,在陕西、山西、甘肃、山东、江苏、安徽、浙江、江西、福建、河南、湖北、湖南、四川、贵州、云南东北部有分布,开发利用前景广阔。但由于华中五味子果实价格不及五味子果实(Schisandrachinensis(Turcz.)Baill.),故常被当作其的代用品,而没有受到人们的关注,直到近几年对华中五味子的研究才开展起来。本论文选取陕西省柞水县华中五味子野生抚育基地的植株为研究对象,运用乙酸乙酯超声提取法和高效液相色谱检测法,对野生华中五味子果实发育阶段木脂素的分布与含量变化进行了研究。旨在了解华中五味子果实生长发育规律,从而确定华中五味子的最佳采收期,为柞水县野生华中五味子资源的开发和利用奠定基础。主要结论如下:
1.确立了高效液相色谱检测华中五味子中木脂素的条件。色谱条件:色谱柱:岛津C18(150×4.6 mm,5μm);流动相:甲醇-水(70:30);流速0.8 mL min~(-1);检测波长:280 nm;柱温:30℃;进样量8μl。方法学考察表明测定方法精密度高、重现性好,可作为华中五味子木脂素含量的检测方法。
2.在华中五味子果实中检测到三种含量较高的木脂素,五味子酯甲、五味子甲素和五味子乙素。显示柞水产华中五味子果实符合药典规定标准,药材品质好。
3.华中五味子植株各部位的木脂素组成和含量差异较大。在果实、叶、多年生茎和一年生茎四个部位中,果实中木脂素种类最多,含量也最丰富,其余三个部位仅含有少量的五味子酯甲。
4.在2007年6-8月间,华中五味子果实的生长发育是一个不连续的过程,分为四个阶段:第一阶段,花后25-40天,为第一个生长阶段;第二阶段,花后40-50天左右,为生长停滞阶段;第三阶段,花后50-75天左右,为第二个生长阶段;第四阶段,花后75天之后,生长基本停止。果实的大小及重量均在花后75天左右时达到最大。
5.华中五味子果实发育阶段,五味子酯甲、五味子甲素和五味子乙素的积累在花后40天时达到最大,之后,五味子酯甲的含量开始急剧减少;五味子乙素的含量则缓慢的减少;五味子甲素的含量基本保持不变,直到花后75天时开始减少。在果实的第一个生长阶段,三种木脂素的含量都是增加的,而在第二个生长阶段,五味子酯甲和五味子乙素的含量减少,五味子甲素的含量基本保持不变。
6.根据华中五味子果实的用途,最佳采收期如下:(1)作为中药使用,最佳采收时期为花后75天左右;(2)用于提取五味子酯甲、五味子甲素和五味子乙素,最佳采收时期分别为花后50天、75天和50天左右。(3)作为水果食用,最佳采收时期为花后75天左右。
Schisandra sphenanthera Rehd.et Wils.(S.sphenanthera) is a kind of perennial lianas which belongs to Schisandra Michx.in Magnoliaceae.Its' dried ripening fruits,which are called nan-wu-wei-zi(Mandarin Chinese,the literal English translation is southern five-taste fruit),are a rich source of lignans with diverse biological activities.The dried fruits of S.sphenanthera have been widely used as a tonic and sedative in China for a long time.S.sphenanthera grow in many provinces of China,such as Shaanxi,Shanxi,Gansu,Anhui,Zhejiang,Jiangxi,Fujian,Henan,Hubei, Hunan,Sichuan,Guizhou,and the northeast part of Yunnan.While the fruits of S.sphenanthera have not attracted enough people's attentions,because the price is lower than Schisandra chinensis (Turcz.) Baill.(another kind of perennial lianas in Schisandra Michx.).So a little of studies have focused on this plant.In this study,a HPLC method for the quantification of four common schisandra lignans,schisandrin,schisantherin A,deoxyschizandrin andγ-schizandrin in S. sphenanthera was developed.And the method was conducted to investigate the lignans changes during S.sphenanthera fruits ripening in four parts of S.sphenanthera,the fruits,the leaves,the multi-year-old stems and the one-year-old stems,expecting to find the develop-pattern of the fruits and determining the best time to harvest,which provide a base-line information regarding these biochemical changes and maximizing the yield for the exploitation of S.sphenanthera in Zhashui in the south of Chin mountain,Shaanxi province in China.The main results were as blow:
1.A rapid and convenient HPLC analysis method had been developed for determination of lignans, schisandrin,schisantherin A,deoxyschizandrin,andγ-schizandrin,in S.sphenanthera.The HPLC system consisted Shimadzu LC-2010(Shimadzu Corp.,Kyoto,Japan) and Shimadzu LCsolution software.Separation was achieved on a Shimadzu C18 column(Phenomenex,150×4.6mm;5μm particle size).The mobile phase consisted of methanol-deionised water(70:30).The flow rate was adjusted to 0.8mL min~(-1),the detection wavelength to 280 nm and 8μm of sample was injected.All separations were performed at 30℃.After the checking of the linearity,the accurately,the repeatability and the precision,the method was showed suitable for the determination of S. sphenanthera.
2.Three lignans were identified in the dried fruits.There were schisantherin A,deoxyschizandrin, andγ-schizandrin.The content of schisantherin A reached the standard of the Chinese Parmacopoeia which indicated that S.sphenanthera from Zhashui was in high grade.
3.The lignans kind and the lignans qualities were vary in different plant organs.In all the four detected parts,fruits,leaves,multi-year-old stems and one-year-old stems,lignans contents were highest in the fruits,not only their kind,but also their qualities.While other three organs,leaves, multi-year-old stems and one-year-old stems,contained only a little of schisantherin A.
4.From Jue to Augest in the year 2007,the development of S.sphenanthera fruits was described as a process of discontinuity,which could be divided into four periods.First period was 25~(th)-around 40~(th) days from anthesis.This was the first growing stage.Second period was about 40~(th)-50~(th) days from anthesis.In this period,the fruits grew very slowly.Third period was 50~(th)-75~(th) days from anthesis or so.This was the second growing stage.Last period was after 75~(th) days from anthesis. The fruits had almost finished growing.Both the weight and the shape of fruits reached their highest points at the 75~(th) days from anthesis.
5.During S.sphenanthera fruit developing,all the three lignans contents accumulated, schisantherin A,deoxyschizandrin,andγ-schizandrin,and reached their highest degrees at the 40~(th) days from anthesis.Then,the schisantherin A content was cut back sharply,theγ-schizandrin content lessened slowly,while the content of deoxyschizandrin maintained almost equably until the 75~(th) days from anthesis when it began to reduce.In the growing stage,the contents of all the three lignans were augmented.In the second growing stage,both the contents of schisantherin A andγ-schizandrin diminished,while the deoxyschizandrin did not vary much.
6.According to the various applications,the S.sphenanthera fruits should be picked up at different time.The best times to harvest were determined as follow.(1) For being used as traditional Chinese medicine,the harvest time was adjusted to around the 75~(th) days from anthesis. (2) For extracting schisantherin A,deoxyschizandrin,andγ-schizandrin,the fruits should be collected at about 50~(th),75~(th),and 50~(th) days from anthesis respectively.(3) For being consuming as commercial fruits,the best time to harvest was determined at 75~(th) days from anthesis.
1.确立了高效液相色谱检测华中五味子中木脂素的条件。色谱条件:色谱柱:岛津C18(150×4.6 mm,5μm);流动相:甲醇-水(70:30);流速0.8 mL min~(-1);检测波长:280 nm;柱温:30℃;进样量8μl。方法学考察表明测定方法精密度高、重现性好,可作为华中五味子木脂素含量的检测方法。
2.在华中五味子果实中检测到三种含量较高的木脂素,五味子酯甲、五味子甲素和五味子乙素。显示柞水产华中五味子果实符合药典规定标准,药材品质好。
3.华中五味子植株各部位的木脂素组成和含量差异较大。在果实、叶、多年生茎和一年生茎四个部位中,果实中木脂素种类最多,含量也最丰富,其余三个部位仅含有少量的五味子酯甲。
4.在2007年6-8月间,华中五味子果实的生长发育是一个不连续的过程,分为四个阶段:第一阶段,花后25-40天,为第一个生长阶段;第二阶段,花后40-50天左右,为生长停滞阶段;第三阶段,花后50-75天左右,为第二个生长阶段;第四阶段,花后75天之后,生长基本停止。果实的大小及重量均在花后75天左右时达到最大。
5.华中五味子果实发育阶段,五味子酯甲、五味子甲素和五味子乙素的积累在花后40天时达到最大,之后,五味子酯甲的含量开始急剧减少;五味子乙素的含量则缓慢的减少;五味子甲素的含量基本保持不变,直到花后75天时开始减少。在果实的第一个生长阶段,三种木脂素的含量都是增加的,而在第二个生长阶段,五味子酯甲和五味子乙素的含量减少,五味子甲素的含量基本保持不变。
6.根据华中五味子果实的用途,最佳采收期如下:(1)作为中药使用,最佳采收时期为花后75天左右;(2)用于提取五味子酯甲、五味子甲素和五味子乙素,最佳采收时期分别为花后50天、75天和50天左右。(3)作为水果食用,最佳采收时期为花后75天左右。
Schisandra sphenanthera Rehd.et Wils.(S.sphenanthera) is a kind of perennial lianas which belongs to Schisandra Michx.in Magnoliaceae.Its' dried ripening fruits,which are called nan-wu-wei-zi(Mandarin Chinese,the literal English translation is southern five-taste fruit),are a rich source of lignans with diverse biological activities.The dried fruits of S.sphenanthera have been widely used as a tonic and sedative in China for a long time.S.sphenanthera grow in many provinces of China,such as Shaanxi,Shanxi,Gansu,Anhui,Zhejiang,Jiangxi,Fujian,Henan,Hubei, Hunan,Sichuan,Guizhou,and the northeast part of Yunnan.While the fruits of S.sphenanthera have not attracted enough people's attentions,because the price is lower than Schisandra chinensis (Turcz.) Baill.(another kind of perennial lianas in Schisandra Michx.).So a little of studies have focused on this plant.In this study,a HPLC method for the quantification of four common schisandra lignans,schisandrin,schisantherin A,deoxyschizandrin andγ-schizandrin in S. sphenanthera was developed.And the method was conducted to investigate the lignans changes during S.sphenanthera fruits ripening in four parts of S.sphenanthera,the fruits,the leaves,the multi-year-old stems and the one-year-old stems,expecting to find the develop-pattern of the fruits and determining the best time to harvest,which provide a base-line information regarding these biochemical changes and maximizing the yield for the exploitation of S.sphenanthera in Zhashui in the south of Chin mountain,Shaanxi province in China.The main results were as blow:
1.A rapid and convenient HPLC analysis method had been developed for determination of lignans, schisandrin,schisantherin A,deoxyschizandrin,andγ-schizandrin,in S.sphenanthera.The HPLC system consisted Shimadzu LC-2010(Shimadzu Corp.,Kyoto,Japan) and Shimadzu LCsolution software.Separation was achieved on a Shimadzu C18 column(Phenomenex,150×4.6mm;5μm particle size).The mobile phase consisted of methanol-deionised water(70:30).The flow rate was adjusted to 0.8mL min~(-1),the detection wavelength to 280 nm and 8μm of sample was injected.All separations were performed at 30℃.After the checking of the linearity,the accurately,the repeatability and the precision,the method was showed suitable for the determination of S. sphenanthera.
2.Three lignans were identified in the dried fruits.There were schisantherin A,deoxyschizandrin, andγ-schizandrin.The content of schisantherin A reached the standard of the Chinese Parmacopoeia which indicated that S.sphenanthera from Zhashui was in high grade.
3.The lignans kind and the lignans qualities were vary in different plant organs.In all the four detected parts,fruits,leaves,multi-year-old stems and one-year-old stems,lignans contents were highest in the fruits,not only their kind,but also their qualities.While other three organs,leaves, multi-year-old stems and one-year-old stems,contained only a little of schisantherin A.
4.From Jue to Augest in the year 2007,the development of S.sphenanthera fruits was described as a process of discontinuity,which could be divided into four periods.First period was 25~(th)-around 40~(th) days from anthesis.This was the first growing stage.Second period was about 40~(th)-50~(th) days from anthesis.In this period,the fruits grew very slowly.Third period was 50~(th)-75~(th) days from anthesis or so.This was the second growing stage.Last period was after 75~(th) days from anthesis. The fruits had almost finished growing.Both the weight and the shape of fruits reached their highest points at the 75~(th) days from anthesis.
5.During S.sphenanthera fruit developing,all the three lignans contents accumulated, schisantherin A,deoxyschizandrin,andγ-schizandrin,and reached their highest degrees at the 40~(th) days from anthesis.Then,the schisantherin A content was cut back sharply,theγ-schizandrin content lessened slowly,while the content of deoxyschizandrin maintained almost equably until the 75~(th) days from anthesis when it began to reduce.In the growing stage,the contents of all the three lignans were augmented.In the second growing stage,both the contents of schisantherin A andγ-schizandrin diminished,while the deoxyschizandrin did not vary much.
6.According to the various applications,the S.sphenanthera fruits should be picked up at different time.The best times to harvest were determined as follow.(1) For being used as traditional Chinese medicine,the harvest time was adjusted to around the 75~(th) days from anthesis. (2) For extracting schisantherin A,deoxyschizandrin,andγ-schizandrin,the fruits should be collected at about 50~(th),75~(th),and 50~(th) days from anthesis respectively.(3) For being consuming as commercial fruits,the best time to harvest was determined at 75~(th) days from anthesis.
引文
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