Study On The Quality Standards And Fingerprints Of The Stem And Leaf Of Abelmoschus Manihot(L)Medic.

Objective:In this paper,we studied the quality standard of Abelmoschus manihot?L?Medic.From the medicinal materials collection and processing,identification,inspection,content determination,fingerprinting items,etc.,the establishment of the specific quality range methods for the proposed version in the 2013 edition of"Guizhou Province,Chinese herbal medicines,ethnic medicines quality standards"Theoretical reference.Method:For the harvest processing and trait control items of the stems and leaves of Abelmoschus manihot?L?Medic.,records are made during collection,processing,and storage;Identification items:?1?microscopic identification:surface production method;?2?Thin layer identification:Take appropriate amount of test solution,spot on polyamide film plate,place in acetone-water-formic acid?4:4:1?developing solvent,take out and dry,spray with 5%AlCl₃-Ethanol solution was developed,dried and observed under a 365 nm UV lamp.Check items:refer to the 15th edition of the Chinese Pharmacopoeia,Item4 of the first method?drying method 0832?for moisture measurement,refer to2302 for ash determination,refer to 2201 items for the determination of leachables and general rule 2321 for heavy metal lead arsenic and mercury determination.Determination items:?1?This experiment used UV-Vis Spectrophotometry to determine the total flavonoid content,the test solution preparation using orthogonal test design,using the total flavonoids as indicators to optimize the conditions,with 40%ethanol as the extraction solvent,Liquid to liquid ratio 0.2 g:25mL,reflux temperature The test solution was prepared at 90? for 100 min and the standard curve was established with rutin as the reference substance.The content of total flavonoids in the stems and leaves of Abelmoschus manihot?L?Medic.was also determined.?2?HPLC method simultaneous determination of rutin,hyperoside,isoquercitrin,test solution was optimized by response surface methodology:55%ethanol,reflux extraction 120min,reflux temperature 80?,methodological study,measured by the standard point method Three-component content.Using the“Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System”published by the National Pharmacopoeia Committee?2012 edition?,common peak test and similarity calculation were performed on the fingerprints of the stems and leaves of Abelmoschus manihot?L?Medic.The common peaks were identified by the comparison of reference substances,and a method for the determination of fingerprints of the stems and leaves of Abelmoschus manihot?L?Medic.was established.Result:The planting,harvesting,and processing of Abelmoschus manihot?L?Medic.medicinal materials are more scientific.In terms of trait description,the description of Abelmoschus manihot?L?Medic.basically conforms to the related literature.Microscopic identification:leaf cross-section surface view of different foliage.The upper epidermis cells are of a single column,square in shape,closely arranged,with visible pores;the lower epidermal cells are smaller and more pores are seen.Thin layer identification items:specific properties,on the polyamide film plate,can show more obvious fluorescent spots at the same position as the reference material.Check items:moisture content between 6.16%and 12.99%,average water content of 8.42%;total ash content between 5.51%and 15.21%,average total ash content of 9.94%;acid insoluble ash content between 0.57%and 2.76%The average acid insoluble ash content was 1.25%;the content of hydrothermal leaching was between 10.47%and 30.17%,the average content was 21.01%;the content of alcohol hot leaching was between 6.49%and 24.77%,and the average content was 14.12%.Due to the low content of heavy metals,they are not included in the standard for the time being.Content determination items:?1?Determination of total flavonoid content in the stems and leaves of Abelmoschus manihot?L?Medic.was determined.The total flavonoids content in each sample ranged from 3.6407 to 32.2402 mg·g^-1.The accumulation of total flavonoids in the stems and leaves of Abelmoschus manihot?L?Medic.from different regions in Guizhou varies greatly.The content of Nanbai and Jinyang from Zunyi was higher,especially in Zunyi Nanbaidong.?2?The methodological validation showed that the linear ranges of rutin,hyperoside and isoquercitrin were0.14¹8.05?g/mL?r=0.9995?,0.19²3.79?g/mL?r=0.9999?,and 0.16¹9.51?g,respectively.The linear relationship was good?r=0.9993?.The average recoveries ranged from 93.73%to 102.10%,91.77%to 102.70%,and 89.72%to 104.55%.The RSD ranged from 0.37%to 1.16%.The contents of rutin,hyperoside and isoquercitrin in the 10 batches of samples ranged from 0.0517to 0.6060 mg·g-1,0.0317 to 0.6811 mg·g-1,and 0.0116 to 0.5064 mg·g-1,respectively.Fingerprint items:There are 7 common peaks in the fingerprints of 10 batches of the stems and leaves of Abelmoschus manihot?L?Medic.;the similarity of the 10 batches of samples:Except for the similarity of S5,S6,and S7?Duyun origin?,the others are all greater than 0.830;The comparison of the reference substance identifies the three components of rutin,hyperoside and isoquercitrin.Using the peak area of 7 common peaks of each sample as a variable,the correlations between the 10 batches of samples and their fingerprints were compared with the stem-leaf ratio of the stems and leaves of Abelmoschus manihot?L?Medic.and plant height by SPSS 19.0 cluster analysis.The samples were divided into three categories,which can distinguish the proportion of the stems and leaves of Abelmoschus manihot?L?Medic.and the height of the plants.Category A is S1,S2,S9,and S10.When harvested,stems and stems account for approximately 70%of stems and stems,and plant height is approximately 40cm.Types B are S5,S6,and S7.About 80%of the plant height is about 60 cm;C species are S3,S4 and S8.The stem and leaf ratio of the stem account for about 60%and the plant height is about 30 cm.Conclusion:For the first time,the research on the systemic quality standards of the stems and leaves of Abelmoschus manihot?L?Medic.was carried out in this study.The experimental study was carried out on the aspects of collection,processing,character,identification,inspection,content,and fingerprints.Thin layer development conditions were optimized for thin layer identification items,and the results demonstrated the superiority of the standards of the stem and leaf of Abelmoschus manihot?L?Medic.in its quality control.In the inspection items,limits are set for moisture content,total ash content,acid-insoluble ash content,and leachate content,and heavy metals are inspected.Since the content of heavy metals?lead,arsenic,and mercury?is lower than the prescribed limit,the inspection of heavy metals is temporary.Not included in this standard.Determination of total flavonoids and multi-components?rutin,hyperoside,isoquercitrin?in the determination of the content,this method is simple,fast,specific,linear,reproducible,precision and recovery rate is good The method can be applied to the quality control of the stems and leaves of Abelmoschus manihot?L?Medic.The determination conditions of the fingerprints of the stem and leaf of Abelmoschus manihot?L?Medic.and the method of fingerprinting of the Abelmoschus manihot?L?Medic.stems and leaves were initially established.