| ObjectiveRosa Roxburghii Radix is the traditionally used minority medicinal plant found in Guizhou province in china which has the efficacy of stomach digestion,convergence and antidiarrheal to treat abdominal pain,diarrhea,etc.At present,we are accelerating the development of a healthy pharmaceutical industry with a focus on Miao medicine,Rosa Roxburghii Radix,a minority medicinal plant,was established on quality improvement studies by Food and Drug Administration of GuiZhou Province.According to the quality standard of Rosa Roxburghii Radix,it is found that the medicinal material standards of this variety are relatively simple,only the shape description,lack of key thin layer identification items,inspection items and content determination items,etc.It is difficult to qualitatively and quantitatively evaluate the status and quality of Rosa Roxburghii Radix.In order to further improve and improve the quality standard for the production of Rosa Roxburghii Radix,and to improve the quality of medicinal materials,the project conducted qualitative and quantitative research on the Rosa Roxburghii Radix,and established the standards and draft standards for Rosa Roxburghii Radix.We hope that the final research results will be a new version of“Chinese Medicinal Materials quality standards of Guizhou Province”.The Rosa roxburghii Tratt is a treasure,and it is used as medicine in many parts.It can not only be used as medicine for medical treatment,but also used for ornamental purposes.It also has broad application prospects in food and beverage,health care products,and cosmetics.In the course of improving the quality standard of Rosa Roxburghii Radix,this study found that the studies of the heavy metal accumulation,migration,risk assessment,source analysis and the status of heavy metal in the soil of Rosa roxburghii Tratt were rarely reported.In addition,the source pollution of the heavy metal is from soil,which has a direct influence on the heavy metal content of the root,leaf,and fruit of the Rosa roxburghii Tratt.Therefore,this article has investigated the contents of heavy metals Pb,As,Hg,Cd,and Cr in the Rosa roxburghii Tratt growing in base of Rosa roxburghii Tratt.The distribution characteristics and pollution risk of heavy metals in different parts of Rosa roxburghii Tratt were studied.The sources of heavy metals enrichment,migration and pollution in the soil-Rosa roxburghii Tratt system were discussed in order to provide reference for quality evaluation,drug safety,and establishment of heavy metal limit standards for Rosa roxburghii Tratt.Method?1?With reference to the“Chinese Pharmacopoeia?2015 Edition?”,the improvement of the quality standard of Rosa Roxburghii Radix includes the following points:?1?Source identification?2?Thin-Layer Chromatography identification?3?Chinese herbal medicine inspection items,including:Moisture determination refers to"Chinese Pharmacopoeia?2015Edition?",Part 4 General Method 0832,Second Method?drying method?;Determination of total ash and acid-insoluble ash refer to"Chinese Pharmacopoeia?2015 Edition?",Part 4 General 2302;Water-soluble Leaching The determination of the substance refers to the fourth general rule 2201 of the Chinese Pharmacopoeia?2015 Edition?;the contents of heavy metals Pb,As,Hg,Cd,and Cr are referenced to GB5009.12-2017,GB5009.11-2014,GB5009.17-2014,GB5009.15-2014 and GB5009.123-2014.?4?Determination of contentDetermination of ellagic acid in Rosa Roxburghii Radix by HPLC?first established?Determination of total polyphenols content in Rosa Roxburghii Radix by UV method?first established??5?Feature Mapping StudyTo establish a preliminary study of the characteristic maps of gallic acid,ferulic acid,ellagic acid and rutin in the extracts of Rosa Roxburghii Radix and to pass the software of.?6?Integrate the above items,and prepare technical requirements in accordance with the quality standards and drafting instructions for the Chinese herbal medicines and ethnic medicines in Guizhou Province,and draw up the draft standards and drafting instructions for the quality standard of the Rosa Roxburghii Radix.?2?Distribution characteristics and risk assessment of heavy metals in roots,stems,leaves,and fruits of Rosa roxburghii Tratt and soil?1?Collect soil and root,stem,leaf and fruit samples of the Rosa roxburghii Tratt from the Rosa roxburghii Tratt planting base?2?Digestion of soil and plant samples according to current national standards and determination of heavy metals?3?According to"GB15618-1995 Environmental quality standard for soils"and"WM/T2-2004 Green standards of medicinal plants and preparations for foreign trade and economy"and"GB 2762-2017 Quantity of pollutants in food",the heavy metal content in soil and plant samples will be initially discussed to investigate pollution characteristics and conditions.?4?Apply soil heavy metal assessment methods?potential ecological hazard index,improved matter element model?and plant heavy metal evaluation methods?enrichment coefficient evaluation method,index evaluation method,human health risk assessment method?to the soil and the thorn pear root,stem,leaf of five kinds of heavy metals in various parts of fruits and vegetables?5?Using mathematical statistics?correlation analysis,principal component analysis?to initially explore the sources of pollutionResult?1?The improvement of the quality standard of Rosa Roxburghii RadixIn the experiment,Thin-Layer Chromatography identification,check items,content determination,and characteristic maps were newly added in the quality standard of Rosa Roxburghii Radix.The research on the improvement of the quality standard of Rosa Roxburghii Radix was completed and a draft quality standard was formed.?1?Identification of the basic source This product is the root of Rosa roxburghii Tratt.f.normalis Rehd.et Wils.and Rosa roxburghii Tratt.?2?The new Thin-Layer chromatography identification method is simple,easy to repeat,and spots are clearly visible.?3?Add and improve all the indicators under the inspection item,complete the sample test of thorn pear root,and initially determine each limit standard.Among them,the water content of 25 batches of Rosa Roxburghii Radix was within the range of 7.38%to 11.95%with an average of 9.59%.The total ash content of 25 batches of Rosa Roxburghii Radix was within the range of 0.41%11.66%,and the average value was 4.03%.The content of acid-insoluble ash in 22 batches of Rosa Roxburghii Radix was in the range of0.10%to 3.33%with an average of 1.03%.The average acid-insoluble ash content of 22 batches of Rosa Roxburghii Radix ranged from 6.65%to 23.23%,and the average value was 14.36%.15 batches of Asparagus in the Rosa Roxburghii Radix were 0.16-0.58 mg·Kg-1,Hg was 0.03-0.12 mg·Kg-1,Cd was 0.01-0.70 mg·Kg-1,and Pb was 0.24-4.04 mg·Kg-1,The content of heavy metals is low.?4?The two methods for determining the contents of ellagic acid and total polyphenols were established.Method 1 For the first time,a total spectrophotometric method for the determination of total polyphenols in the Rosa Roxburghii Radix was established.Gallic acid was used as a control.The detection wavelength was766 nm.The total polyphenol content of 15 batches of thorn pear root samples was 7.54%to 14.04%,with an average of 10.70%.RSD is 21.24%.Method 2 The RP-HPLC determination of ellagic acid was established for the first time.The COSMOSIL 5C18?4.6 ID×250 mm,5?m?column was used for the isocratic elution with acetonitrile-0.1%aqueous solution of phosphoric acid as the mobile phase.The detection wavelength was 254 nm;Volume flow 1mL·min-1;column temperature 30°C;injection volume 10?L.The linearity of ellagic acid was good in the range of 2.43-21.87?g·mL-1.The average recovery rate was 105.83%and the RSD was 3.23%.The content of ellagic acid in 20 samples of Rosa roxburghii Radix samples ranged from 0.20to 1.48 mg·g-1 with an average value of 0.76 mg·g-1.?5?Established specific chromoatograms for the extraction of Rosa Roxburghii Radix by HPLC.Twelve common peaks were identified by TCM fingerprint similarity evaluation system software?2012 version?.The similarity of 12 batches of specific chromoatograms reached 0.8 or more.Peaks Nos.4,10,11 and 12 were determined by comparison of reference materials.The four components were gallic acid,ferulic acid,ellagic acid,and rutin,respectively,and this components were first detected in the Rosa Roxburghii Radix.?6?Complete the draft quality standards and drafting instructions.?2?Distribution characteristics and risk assessment of heavy metals in roots,stems,leaves,and fruits of Rosa roxburghii Tratt and soil?1?The heavy metals Pb,As,Hg,and Cd in the 45 batches of soil samples from the four Rosa roxburghii Tratt bases were all within the limits of the national secondary soil standard,and were in conformity with the requirements for heavy metals in base soils for the cultivation of Chinese herbal medicines.?2?The 131 samples of Rosa roxburghii Tratt containing 15 batches of root,48 batches of stem,48 batches of leaf,20 batches of fruit detected,were based on the limits of heavy metals in the“Green standards of medicinal plants and preparations for foreign trade and economy”?WM/T2-2004?.The sample pass rates were:%,93.5%for C.oleifera,80%for C.oleifera,and60.5%for C.oleifera.?3?The distribution characteristics of heavy metals in different parts of Rosa roxburghii Tratt were stem>leaf>root>fruit.From the distribution characteristics of heavy metals,the pollution status of heavy metals was Cr>Cd>Pb>Hg>As;From the point of view of over-standard elements,Cr in the roots,stems and leaves of Rosa roxburghii Tratt all exceeded the standard.?4?The single factor pollution index of each heavy metal in the fruit,root,stem and leaf of the thorn pear was Cr>Cd>Pb>Hg>As.The comprehensive pollution index of each part of Rosa roxburghii Tratt was root>stem>leaf>fruit,the enrichment coefficient was Cd>Hg>Pb?As?Cr.?5?The overall soil condition of the four bases is good,and they are in lighter pollution levels.A small amount of samples have Hg and Cd pollution,of which the potential ecological risk of Hg is strong.?6?The Cd polluted by Rosa roxburghii Tratt mainly comes from soil,and Hg,Pb,As,Cr may be related to atmospheric deposition and agricultural activities.?7?Base soils Cd and Cr are derived from parent materials,and As and Pb may originate from agricultural activities,while Hg pollution may be closely related to atmospheric deposition.Conclusions and highlightsThis study carried out a preliminary exploration of the Rosa Roxburghii Radix in terms of thin-layer chromatographic identification,inspection items,content determination,and characteristic maps to achieve the purpose of improving the quality standard of the Rosa Roxburghii Radix,and provided a theoretical basis for improving the quality standard of the Rosa Roxburghii Radix.Based on the field investigation and indoor analysis,the same surface soil of Rosa roxburghii Tratt and different parts of Rosa roxburghii Tratt was collected.The contents of heavy metals Pb,As,Hg,Cd and Cr in different parts of Rosa roxburghii Tratt and soils were determined by atomic absorption spectroscopy and atomic fluorescence spectrometer respectively.Then the heavy metals in the soil and various parts of the Rosa roxburghii Tratt were preliminarily explored according to national standards.The characteristics and circumstances of the pollution were investigated;the evaluation methods of soil heavy metals?potential ecological hazard index,improved matter element model?and plant heavy metal evaluation method?enrichment coefficient evaluation method,exponential evaluation method,and human health risk assessment method?were used.Methods were used to evaluate the contents of five heavy metals in the soil,roots,stems,leaves,and fruits of each sample;mathematical statistics?correlation analysis,principal component analysis?was used to explore the sources of pollution. |
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