Study On The Optimal Harvest Time Of Schizonepeta Tenuifolia Based On Chemical Composition And Biologicai Activity

In the cultivation of Chinese herbal medicines to harvest growing,its composition is a dynamic,complex changing process,harvesting will directly affect the yield,quality and efficiency of harvesting herbs.Schizonepetae herba,is the whole dry part on the ground of Schizonepeta tenuifolia Briq.from Labiatae.It is slightly warm in nature?acrid flavor and go back to lung and liver.It has the effects of relieving superficies,promoting eruptions and slaking sore and mainly used in cold,headache,measles,rubella,sores ulceration and so on in Clinical."Chinese Pharmacopoeia" of 2010 edition provisions that the summer and autumn are suitable harvest while flowers ceiling and tapping down spike green.The content of volatile oil is regard as the main indicator to evaluate the quality of Schizonepeta tenuifolia.However,biosynthesis still ongoing in the harvest period,different harvest periods and conditions are likely to affect the composition ratio of volatile oil medicinal ingredient in all kinds of compounds and each other,and it can be result the different in quality,pharmacodynamic and toxicological.Therefore,in order to determine the appropriate scientific and reasonable harvest time,except the content of volatile oil must be understand in different harvest times,the main components content of the volatile oil changing and differences of its pharmacodynamic studies are necessary.The paper selected Schizonepeta tenuifolia from the main producing areas as a research object,by Characters,determination of the content of volatile oil,the main component changing of volatile oil,the anti-inflammatory efficacy of volatile oil,and anti-viral activity,namely sensory evaluation-chemical evaluation-biological evaluation of a combination of comprehensive determination of optimum harvest.The studies in this paper mainly include the following sections:Part one:Literature reviewAfter having reviewed relative books and literatures,we summarize a review of the chemical composition,pharmacological effects,the harvest times and the status and direction of development of Chinese medicine quality control,to support the new research direction,the feasible experimental program and examine indicators.Part two:The collecting and determining the content of volatile oil of Schizonepeta tenuifolia in different harvesting timesFrom Flowers early to fruit,were taken once every seven days observing its appearance and shape,manual dispensing stem,leaf,spike and calculate the percentage.Proportionally weight amount of stem,leaf,spike,and determine of the content of volatile oil.The results showed that,Schizonepeta tenuifolia the proportion of stem volatility is not too large,generally about 50%,proportion of leaf?spike fluctuations in different harvest times;The overall trend of volatile oil content is decreased,which In the harvesting point 1 volatile oil content up to 2.191%(ml/g).So the characters and content of volatile oil have a big difference with the different of harvesting times.According to the "Chinese Pharmacopoeia" of 2010 edition provides the optimun harvesting point of Schizonepeta tenuifolia is flowers meeting the ceiling and spike becoming green,that meeting the requirements are:3,4,5,corresponding harvesting time are 2012.09.06,2012.09.13,2012.09.20.Traditionally experience,higher levels of volatile oil as the optimum harvesting time,the appropriate harvest point of Schizonepeta tenuifolia is 1,2,3,corresponding harvesting time are 2012.08.23,2012.08.30,2012.09.06,the harvesting point 1,that harvesting time 2012.08.23 is better harvesting time.Part three:Determining the content of main components of volatile oil of Schizonepeta tenuifolia in differenting harvest timesBy GC-MS simultaneous determinate the content of main components of volatile oil(1-octen-3-alcohol,3-octanone,?-myrcene,d-limonene,(-)-menthone,(+)-menthofuran,(+)-pulegone,?-caryophyllene).The results showed that,Schizonepeta tenuifolia in different harvest times,the content of 1-octen-3-alcohol firstly increases and then gradually reduces;the content of ?-caryophyllene,3-octanone,?-myrcene,d-limonene,(+)-menthofuran in the overall trend are gradually reduced.The content of(+)-pulegone firstly reduces and then increases,and reached the maximum content was 860.07mg/g in harvesting time 7;the content changing of(-)-menthone has a opposite trend with(+)-pulegone,and reached the maximum content was 322.22mg/g in harvesting time 3.In summary,the content of main volatile components of Schizonepeta tenuifolia are constantly changing with the different of harvest times,origin.Therefore,the development of appropriate harvest time,will have a direct bearing on the quality of Schizonepeta tenuifolia.Part four:Determining the effects of anti-inflammatory and The anlylisis spectrum-effect relationshipDetermining the effects of anti-inflammatory by carrageenan-induced pleurisy in rats from volatile oil of Schizonepeta tenuifolia in different harvesting times:Using carrageenan-induced pleurisy in rats,with the volume of exudation,the number of white blood cells,protein content,MDA content,MPO activity and lung histopathology as evaluation,study on the effects of anti-inflammatory to carrageenan-induced pleurisy in rats from volatile oil.The results showed that,the volatile oil of Schizonepeta tenuifolia has the strongest inhibition when the harvesting times are 1,2,3,7,8.The volatile oil of Schizonepeta tenuifolia in other harvest times also showed different degree inhibition on carrageenan-induced pleurisy in rats.By the effects of anti-inflammatory as evaluation the results showed that:the harvesting point of Schizonepeta tenuifolia are 2,3,7,8,corresponding harvesting time are 2012.08.23,2012.08.30,2012.09.06,2012.10.04,2012.10.11.The anlylisis spectrum-effect relationship between the volatile components and anti-inflammatory effects:Using gray correlation analysis method to study the degree of association between the content changing of main volatile components(1-octen-3-alcohol,3-octanone,?-myrcene,d-limonene,(-)-menthone,(+)-menthofuran,(+)-pulegone,?-caryophyllene)of Schizonepeta tenuifolia in different harvest times and the inflammatory markers,including the volume of exudation,the number of white blood cells,protein content,MDA content,MPO activity.The results showed that,the order of average association level is(+)-Pulegone>P-caryophyllene>d-limonene>?-myrcene>1-octen-3-ol>3-Octanone>(-)-menthone>(+)-menthofuran;and(+)-pulegone,?-caryophyllene,d-limonene,?-myrcene have the larger degree of association and can be considered they have large contribution to reduce inflammatory markers.And the degree of association between(+)-pulegone,?-caryophyllene and inflammatory markers was significantly higher than other compounds,thus we concluded that these two components in Schizonepeta tenuifolia may be an important material basis of anti-inflammatory effects.The substance of volatile oil plays the efficacy of anti-inflammatory in addition to(+)-pulegone,the low levels of other compounds may also play a key role.Selecting the relative content of two components-(+)-pulegone,?-caryophyllene,and combinating the efficacy of anti-inflammatory at different harvesting times,evaluate the appropriate harvesting time.In the harvesting point 3,the relative content of(+)-pulegone and ?-caryophyllene reach maximum,and the efficacy of anti-inflammatory is remarkable,so select harvesting point 3 as a better harvest time.Corresponding harvesting time is 2012.09.06.The Chinese mdicine of Schizonepeta tenuifolia has complex chemical composition,exploring clear what kind or what kinds of compounds in clinical role to play in what proportion will be a problem for a long time.Therefore,the experiment mainly from the perspective of biological activity study the optimal harvesting time of Schizonepeta tenuifolia.Part five:Studying antiviral activity of Schizonepeta tenuifoliaDetermining antiviral activity of Schizonepeta tenuifolia based on NA:Using a neuraminidase inhibitor screening kits,firstly establish a standard curve of neuraminidase activity,and then calculate the sample for neuraminidase inhibition percentage witch relative the standard curve.And then do the inhibition rate-concentration curve of the sample to calculate IC50 of the sample.The results showed that,IC50 of Schizonepeta tenuifolia from Hebei firstly increases and then gradually reduces,that antiviral activity firstly increases and then gradually reduces,when the harvest time is 2012.09.06,IC50=54.37 mg·ml-1,the antiviral activity reach strongest.So Schizonepeta tenuifolia at different harvesting times all have some degree of antiviral activity,the antiviral activity of Schizonepeta tenuifolia is different with the different of harvest times.By the antiviral activity as evaluation the results showed that:The appropriate harvesting point of Schizonepeta tenuifolia are 2,3,4,corresponding harvesting time are 2012.08.30,2012.09.06,2012.09.13;the harvesting point 3,that harvesting time 2012.09.06 is better harvesting time.Determining the hemagglutination activity of Schizonepeta tenuifolia:Establish a method that can determinate the antiviral activity of Schizonepeta tenuifolia,and rely on the method to inspect antiviral activity of Schizonepeta tenuifolia in different harvest times.Coagulation experiment using a microtiter plate,after optimization and validation methodology,determination of antiviral activity limits established based on hemagglutination of Schizonepeta tenuifolia,its specificity,stability,precision,which all adhere to the relevant requirements of pharmaceutical bioassays requirements;The results showed that,the hemagglutination activity in the overall trend are gradually reduced.When the harvesting is 1,2,3,the hemagglutination activity is better.When the harvesting is 3,highest dilution positive is 1:210,the hemagglutination activity reached its strongest.Therefore,based on hemagglutination activity assay of Schizonepeta tenuifolia showed that,the appropriate harvesting point of Schizonepeta tenuifolia are 1,2,3,corresponding harvesting time are 2012.08.23,2012.08.30,2012.09,06;the harvesting point 3,that harvesting time 2012.09.06 is better harvesting time.Part six:Determining the appropriate harvesting of Schizonepeta tenuifoliaThe summary of results based on different indicators evaluating the optimum harvesting time of Schizonepeta tenuifoliaNote:The underlined that a better harvest time under indicatorsAs can be seen from the table,Schizonepeta tenuifolia more appropriate harvesting times are in late August or early September.Which Schizonepeta tenuifolia in late August at the highest point of the volatile oil content,its anti-viral activity is good;Schizonepeta tenuifolia oil content in early September and harvested in early August,though,but still at a high point,its relative content of pulegone and P-caryophyllene reach maximum,its anti-inflammatory activity,anti-viral activity all were good,Schizonepeta tenuifolia adopted in early September compliance with the "Chinese Pharmacopoeia" of 2010 edition Schizonepeta tenuifolia items bloom ceiling,green spike requirements;Therefore,the above analysis,this paper argues that early September is the best harvesting time of Schizonepeta tenuifolia.