| Rumex chalepensis Mill,which root and rhizome belonging to the genus Rumex of Polygonaceae is a kind of traditional Chinese medicine.The medicine has bitter taste and coldness.It has the functions of clearing away heat and detoxifying,breaking new sputum,reducing swelling and pain,stopping bleeding,laxative,and killing insects.It is used for hemostasis and treatment of dysentery.Modern clinical application believes that it has the effect of regulating qi and activating blood circulation,breaking new sputum,and reducing swelling and muscle growth.Its single-drug or compound treatment of Helicobacter pylori(Hp)-related gastritis,peptic ulcer,duodenal bulb ulcer,etc.have achieved good results.In the 1980s,the Jingzhou Traditional Chinese Medicine Hospital of Hubei Province had made a Rumex chalepensis Mill.granule,which had a good curative effect in the clinical treatment of Hp-associated gastritis.However,the basic research of Rumex chalepensis Mill,is relatively weak.There is no local or national standard.The quality is difficult to control.The preparation has not undergone strict process screening,and it is difficult to exert maximum efficacy.The toxicity and pharmacodynamic research was almost blank.Therefore,it is necessary to carry out comprehensive basic research on the Rumex chalepensis Mill,that has been clinically applied for more than 30 years.This subject was to study the chemical composition,quality standard,preparation,acute toxicity and pharmacological activity of gastric acid pepsin.The main contents and results were as follows:1 The original identification of Rumex chalepensis MillThe sample for identification of Rumex chalepensis Mill was from Zhangzhuangpu Town,Gong’an County,Jingzhou City,Hubei Province.There were 2 batches,the first batch had roots,stems,leaves and flowers:the second batch had roots,stems,leaves and fruits.Professor Wang Zhenyue of Heilongjiang University of Traditional Chinese Medicine based on plant morphology and anatomical similarity,according to Volume 25(1)of Chinese Flora,identified the dried roots and rhizomes of Rumex chalepensis Mill.2 Chemical Composition Analysis of Rumex chalepensis Mill.UPLC-Q-TOF-MS technology was used to analyze the chemical components that may be contained in the Rumex chalepensis Mill.Results A total of 18 compounds were identified,including 3 ruthenium compounds,3 free guanidine compounds,4 naphthoquinone compounds,7 flavonoids and 1 lignan.The results of this study laid the foundation for the clarification of quality control and pharmacodynamic mechanisms.3 Establishment of Quality Standard for Rumex chalepensis Mill.3.1 TLC Identification of Rumex chalepensis Mill.Hin layer identification was performed on five components in the Rumex chalepensis Mill..The silica gel G plate was used as the stationary phase,and ethyl acetate-chloroform-acetone-methanol-water(1:5:2.5:1.5:0.4)was used as the developing solvent to detect the chrysophanol-8-O-β-D-glucoside at 365 nm.;using silica gel G plate as stationary phase,petroleum ether(60～90 ℃).ethyl acetate-acetone-formic acid(6:1:1:0.5)as developing agent,in UV The emodin,nepodin,chrysophanol,and physcion were detected at 365 nm.The thin layer map showed that the test sample shows spots of the same color at the same position as the standard sample,and the components of the sample were well separated,Rf chrysophanol-8-O-β-D-glucoside=0.59,Rf emodin=0.26,Rf nepodin=0.46,Rf chrysophanol=0.54,Rf physcion=0.62,Rf values were between 0.2 and 0.8.The above five components could be used as a thin layer identification.3.2 Established a method for determining the liquid phase content of Rumex chalepensis Mill.,and clarified its growth period and harvest time.A method for the determination of Rumex chalepensis Mill,was established by high performance liquid chromatography.By examining the column,mobile phase,and detection wavelength,the determination conditions were as follows:the chromatographic conditions were:Agilent Extend-C18(250 × 4.6 mm,5 μm),and the diode array detector at 254 nm was detected.The mobile phase is:A methanol,B 0.1%formic acid,gradient elution,0～14 min,45%～60%A,14～25 min,60%～86%A,25～30 min,86%～100%A,30～35 min,100%A.The flow rate was 1 mL/min,the column temperature was 25℃,and the injection volume was 5 μL.By screening the solution extraction method,solvent,solvent volume and extraction time,the preparation method of the solution was determined as follows:1.0 g of Rumex chalepensis Mill,powder(passing 50 mesh sieve),adding 40 mL of methanol,and ultrasonic extraction for 1 h.And methodological investigation,standard curve,precision,stability,repeatability,sample recovery and recovery meet the requirements of content determination.The content of the main components contained in different growth years and different harvesting seasons,and the Rumex chalepensis Mill,in the different blocks was measured.The results showed that 16 batches of Rumex chalepensis Mill.chrysophanol-8-O-β-D-glucoside,emodin-8-β-D-glucopyranoside,nepodin emodin,chrysophanol,physcion The content of them were determined to be 0.06%to 0.71%,0 to 0.27%,0.10%to 0.65%,0.01%to 0.06%,0.07 to 0.43%,and 0.01%to 0.04%,respectively.The above six components could be used as indicators for controlling the content of Rumex chalepensis Mill.At the same time,for the comparison of different growth years,different harvesting periods and different plots of samples,it was considered that the optimal harvesting period of Rumex chalepensis Mill,was 2 years,which was suitable for collection in early spring or late summer and early autumn.3.3 Established Rumex chalepensis Mill,fingerprintFingerprint analysis of 16 batches of Rumex chalepensis Mill,was carried out by liquid chromatography.The results showed that 16 batches of Rumex chalepensis Mill,and 10 common peaks were obtained at 254 nm.The chromatographic peaks were well separated and the average similarity was 0.91.Comparing and analyzing the content of different plots,different growth years and different harvesting seasons,it was found that the similarity of HPLC fingerprints of Rumex chalepensis Mill.,but they were all greater than 90%.The fingerprint of the Rumex chalepensis Mill.established in this paper was characteristic and specific,and can be used to comprehensively control the quality of Rumex chalepensis Mill..4 Study on the preparation of Rumex chalepensis Mill.4.1 Extraction of Rumex chalepensis Mill,and Optimization of Optimum Technological ParametersL9(3)4 orthogonal design and single factor investigation were carried out.The total extraction rate and inhibition zone of the main components of Rumex chalepensis Mill.were used as indicators to study the water extraction and alcohol precipitation process and alcohol extraction process of Rumex chalepensis Mill.At the same time according to Jingzhou Hospital Rumex chalepensis Mill,granules production process to prepare Rumex chalepensis Mill,samples.The experimental results showed that the optimal alcohol extraction process of Rumex chalepensis Mill,was extracted with 6 times 75%ethanol for 90 min,and extracted for 3 times.The total extraction rate of nepodin and anthraquinone was 0.73%,which is significantly higher than that of water extraction.0.24%of the process,0.28%of the water extraction and alcohol precipitation process and 0.24%of the Jingzhou Hospital granule process,the transfer rate of the six components is 84.11%,which is higher than the 27.68%of the water extraction process,33.08%of the water extraction and alcohol precipitation process and Jingzhou The original granule process is 27.68%.Therefore,the optimum extraction technology of Rumex chalepensis Mill,was to add 6 times of 75%ethanol and reflux for 90 minutes and extract for 3 times.4.2 Preparation of Rumex chalepensis Mill.CapsulesAccording to the 2015 edition of the Chinese Pharmacopoeia(General Rule 0103),Ethanol extract capsule excipient Rumex chalepensis Mill.was prepared.The moisture content,the difference in the volume,the time limit of disintegration and the stability were all in accordance with the requirements of the Chinese Pharmacopoeia.Excipient capsules:0.45 g/grain,taking method:3 times/day,3 capsules/time.4.3 Established HPLC method for determination of Rumex chalepensis Mill.A method for the determination of Rumex chalepensis Mill.Capsules by high performance liquid chromatography was established.The contents of Rumex chalepensis Mill,capsule mainly contain chrysophanol-8-O-β-D-glucoside,emodin-8-β-D-glucopyranoside,nepodin,emodin,chrysophanol and physcion.The average contents of six components in three batches of Rumex chalepensis Mill.capsules without excipients were 3.20%,0.40%,0.23%,0.10%,1.03%,0.25%,and the total content was 5.21%.40%,0.31%,0.17%,0.07%,0.75%,0.19%,the total content was 4.89%.The results provided a scientific basis for the quality control of Rumex chalepensis Mill,capsules.4.4 Established a thin layer identification method for Rumex chalepensis Mill.CapsulesTLC identification of five components in Rumex chalepensis Mill,capsule was carried out.The identification was carried out with reference to the TLC identification method of Rumex chalepensis Mill..TLC showed that the samples showed the same color of fluorescent spots at the same position as the control.Meanwhile,the separation degree of each fluorescent spots was good.Rf.chrysophanol-8-O-β-D-glucoside=O.59,Rf emodin 0.26,Rf nepodin=0.46,Rf chrysophanol=0.54,Rf physcion=0.62,Rf valueswere between 0.2 and 0.8.The method can be used as a thin layer identification method for five components of Rumex chalepensis Mill.5 Study on Acute Toxicity of Rumex chalepensis Mill.In the absence of LDso,the maximum tolerated dose was not obtained.The results showed that the amount of gastric perfusion in mice was 250 times that of humans,which was much greater than 100 times,indicating that the toxicity of Rumex chalepensis Mill.75%ethanol extract was very small.6 Effect of Rumex chalepensis Mill,on gastric acid pepsin in rats with pyloric ligation gastric ulcerPyloric acid ligation was used to investigate the effects of 75%ethanol extracts from Rumex chalepensis Mill,on gastric acid and pepsin in rats.The experimental results showed that each of the drug-administered groups had the effect of reducing the total acidity of gastric juice,the total acid output per hour and the activity of gastric tissue protease in rats with gastric ulcer.The high-dose group of Rumex chalepensis Mill.combined with omeprazole(western medicine)The ability to inhibit gastric acid secretion is the strongest in each drug-administered group,and its inhibitory effect on gastric acid and pepsin is superior to the positive drug omeprazole,and superior to the single-flavored Chinese medicine Rumex chalepensis Mill.,indicating that Rumex chalepensis Mill.The high dose group combined with omeprazole has synergistic effects. |
