Changes Of Chemical Components And Quality Control Of Morning Glory Seeds Before And After Frying

Semen pharbitidis is the dried mature seeds of Convolvulaceae plant Pharbitis nil(L.)Choisy or Pharbitis purpurea(L.)Voigt.It first appeared in the book Mingyi Bielu,and also known as Hei Chou,Bai Chou,Er Chou;etc.The character of it is cold,bitter and poisonous,into the lung,kidney and large intestine meridian.It can cause drastic purgation for clearing away retained fluid,and kill worms.It is used for therapy of ansarca with cinstipation and oliguria,dyspnea and cough caused by retained fluid,abdominal pain due to intestinal parasitosis,taeniasis.Pharbitic not only has a long history of medicinal preparation,but also has a long history of processing.Its processing first appeared in the Lei Gong Pao Zhi Lun.Ancient processing methods are commonly used with wine,salt,stir-frying etc.The Chinese Pharmacopoeia of 2010 edition recorded the semen pharbitidis processed products is stir-fried semen pharbitidis,and provinces processing norms is mostly used in stir-frying method.Study shows that Pharbitic raw materials commonly used for induce diuresis to alleviate edema,insecticide.After stir-fried,its toxicity will reduce,efficacy can be tempered,and become easily broken and the active ingredient dissolved out,so it is more commonly used for digestion.Therefore,Pharbitic and stir-fried semen pharbitidis have different effects in clinical application.At present,there are few studies on semen pharbitidis,most of them are focused on the chemical components and pharmacological aspects.Clinical often used stir-fried semen pharbitidis,but there is a lack of relevant research on stir-fried semen pharbitidis pieces at present.The standard of semen pharbitidis in ’China pharmacopoeia’(2015 edition)is the same to in ’China pharmacopoeia’(2010 edition).Therefore,this research is necessary.Objective:1.Study on the chemical constituents of Semen pharbitidis,and establish the quality evaluation method.2.Analysis the principle of stir-frying and stir-frying technology.3.Study on the chemical constituents of stir-fried Semen pharbitidis,and establish the quality evaluation method.Method:1.Chemistry research of semen pharbitidis and stir-fried semen pharbitidis.Collect the semen pharbitidis come from Hebei,and processed into stir-fried semen pharbitidis.By using HPLC-MS method to analyze the chemical composition of semen pharbitidis before and after fried and the corresponding structures were speculated.2.Using the multi index comprehensive weighted scoring method to optimize the best stir-flying process of semen pharbitidis.Using single factor investigation method to inspect different stir-frying time.We screened the optium processing methods mainly through the appearance,the assay of the content of water-soluble extractive and ethanol-soluble extractive.By giving the correspond coefficient to each index weight to calculate the comprehensive score for each processing level samples,then optimize best frying process.3.Research of specific chromatogram.Using HPLC method to establish the specific chromatogram of semen pharbitidis and stir-fried semen pharbitidis.The conditions were as followed:column:Agilent Eclipse XDB-C18(4.6×250mm,5μm),detection wavelength:296nm,column temperature:25℃,flow rate 1.0mL/min,injection volume:10μL,with the 0.1%formic acid as the mobile phase A,methanol as mobile phase B for gradient elution.4.Research of extract content.According to the determination method from 2010 edition of ’Chinese Pharmacopoeia’ appendix X A to determine the alcohol extract content and water extract content of herbs and pieces.Result:1.Through HPLC and HPLC-MS,a total of 16 compounds were identified,they are phenolic acids,flavonoids,phenolic acid lactones.This compounds are 5-CQA,3-CQA,4-CQA,CA,5-pCoQA,5-FQA,FA,3-CQAL,4-CQAL,3,4-DiCQAL,3,4-DiCQA,3,5-DiCQA,4,5-DiCQA and so on.Three components of 3-CQAL,4-CQAL,3,4-DiCQAL were found in the mass spectrum of stir-fried semen pharbitidis,and not found in semen pharbitidis,So they are the peculiar compositions of stir-fried semen pharbitidis2.By measuring the appearance,alcohol extract and water extract of different levels stir-fried samples,the result showed that when the frying time was 10-12min,the comprehensive score was the highest,so the best process is as follows:take net semen pharbitidis 200g,stir-fried 10-12 min under 200℃,then remove out and let cool.3.Analysis the HPLC specific chromatogram of different origins medicinal materials,we found that there have a total of 17 common peaks,seven of them were identified.Stir-fried samples have a total of 21 common peaks,seven of them were identified.Compared the two specific chromatograms we can know,the common peaks of stir-fried samples than semen pharbitidis increased by 4.The content of CA,3-CQA,3,5-DiCQA reduced and the content of 5-CQA,4-CQA,3,4-DiCQA,4,5-DiCQA increased.Through the study of stability,repeatability and precision etc on specific chromatogram we can conclude that main common peaks RSD of relative retention time and relative peak area are consistent with the requirements,it indicates that the established specific chromatogram meets the requirements of the establishment of fingerprint technology and can be used for qualitative analysis.4.The extract content of semen pharbitidis and stir-fried semen pharbitidis from different origins were determined.The results showed that the 10 batches herbs in alcohol extract(maceration)content range from 17.24%to 20.51%,water soluble extract(hot dip)content range from 33.57%to 37.04%;the 10 batches stir-fried semen pharbitidis in alcohol extract(maceration)content range from 16.36%to 17.33%,water soluble extract(hot dip)content range from 39.12%to 42.03%.After comparison,the two groups both have no obvious difference among the 10 batches,showed that the different origins have little effect on the seed extract content;Comparison of semen pharbitidis and stir-fried semen pharbitidis content shows that stir-fried semen pharbitidis alcohol extractive content was decreased,mean from 19.05%down to 16.77%,the content of water extracts increased,mean from 35.42%rise to 40.22%.Because the water extract is also part of effective component of medicinal materials and stir-fried samples,it is recommended that we should increase the control of water extract content of semen pharbitidis and stir-fried semen pharbitidis.So it is recommended that the water soluble extracts content of semen pharbitidis in dried meter is not lower than 29.75%,the content of stir-fried semen pharbitidis in dried meter is not less than 34.25%.Conclusion:1.This study is for the first time to detect out quercetin and quercetin glucoside from semen pharbitidis,but also for the first time to detect out flavonoids from semen pharbitidis;By comparing the chemical composition of semen pharbitidis and stir-fried products we can know,the chemical components and contents are all changed before and after stir-fried.In terms of species,three components of 3-CQAL,4-CQAL,3,4-DiCQAL were found in the mass spectrum of stir-fried semen pharbitidis,and not found in semen pharbitidis,So they are the peculiar compositions of stir-fried semen pharbitidis;In terms of component content changes,for stir-fried semen pharbitidis,the content of CA,3-CQA,3,5-DiCQA compared with raw medicinal materials decreased,while the content of 5-CQA,4-CQA,3,4-DiCQA,4,5-DiCQA increased.This method can be more comprehensive understanding of the changing rule of the main chemical composition before and after stir-fried,for the semen pharbitidis processing mechanism and stir-fried samples quality standards research provides basic chemical information.This can provide basic information for the researching work on process mechanism and quality standard of decoction pieces.2.Validation test showed that the optimized process is stable;therefore,this stir-frying process is reasonable.3.Analysis the HPLC specific chromatogram of different origins medicinal materials,found that the overall chromatogram looks almost the same,the relative retention time and relative peak area of characteristic peaks have little difference,indicating that each semen pharbitidis from different origins contains similar chemical composition,the medicinal materials quality is stable.For ten groups of stir-fried semen pharbitidis,the relative retention time and relative peak area of characteristic peaks also had little difference.Compared the HPLC specific chromatogram and the relative peak area of the semen pharbitidis before and after stir-fried we can know,after stir-fried the specific chromatogram changed significantly.Therefore,HPLC specific chromatogram can quickly and efficiently identify the semen pharbitidis herbs and stir-fried pieces,and can be used for the overall quality evaluation,so as to provide a scientific basis for effectively control the medicinal materials inner quality standard.4.Comparative extract content shows that semen pharbitidis,stir-fried semen pharbitidis are not have obvious differences among batches,and indicating that origin of pharbitis seed has little effect on extract content;compared semen pharbitidis and stir-fried semen pharbitidis extract results shows,alcohol extract content decreased and water extract content increased after stir-fried.Studies have pointed out that after fried semen pharbitidis volatile oil content decreased to produce aroma,so the alcohol extract content decrease may be related to the decrease of these substances;In addition,stir-fried semen pharbitidis become easily broken and the dissolute of effective components become easily too,so the water extract content increased,this is consistent with the traditional ’feng zi bi chao’ view.