Herbal Textual Research on Zingiberis Rhizoma Recens in Famous Classical Formulas and Its Quality Evaluation / 中国实验方剂学杂志

ObjectiveTo provide references for the selection of Zingiberis Rhizoma Recens on the research of famous classical formulas and the reasonable uses for medicines and foods through herbal textural research and quality analysis of Zingiberis Rhizoma Recens from main producing areas in China. MethodBy consulting the ancient and modern literature， the name， origin， producing areas， harvest time， processing methods of Zingiberis Rhizoma Recens were summarized. According to the 2020 edition of Chinese Pharmacopoeia， the contents of 6-gingerol， 8-gingerol， 10-gingerol， and volatile oil in Zingiberis Rhizoma Recens samples were determined. ResultHerbal textural research indicated that medicinal Zingiberis Rhizoma Recens originated from the fresh rhizome of Zingiber officinale. Before Tang dynasty， Zingiberis Rhizoma Recens produced in Sichuan was the best. In the Song dynasty， Zingiberis Rhizoma Recens produced in Sichuan， Zhejiang， and Anhui was of excellent quality. The cultivation of Zingiberis Rhizoma Recens in Shandong developed during the Ming and Qing dynasties. From ancient times to the present， the harvest period extended from the autumnal equinox to the winter solstice. Quality evaluation standards of Zingiberis Rhizoma Recens were essentially the same in ancient and present documents， as those with little gluten or gluten-free and strong pungency were preferred. After determination， the contents of 6-gingerol， 8-gingerol， and 10-gingerol in 44 samples were qualified in 27 samples， with a qualified rate of 61.4%. Among them， 17 samples were unqualified in the total contents of 8-gingerol and 10-gingerol. Among these qualified samples， the content of 6-gingerol ranged from 0.067% to 0.255%， and the total contents of 8-gingerol and 10-gingerol ranged from 0.040% to 0.131%. The content of volatile oil in 36 samples were qualified in 33 samples， with a qualified rate of 91.7%. Among the qualified samples， the content of volatile oil ranged from 0.175% to 0.410%. ConclusionZingiberis Rhizoma Recens has been used as medicines and foods since ancient times， and the genuine producing areas are consistent in ancient and present times， while the quality of the products， especially the medicinal Zingiberis Rhizoma Recens， should be monitored. Medicinal Zingiberis Rhizoma Recens planted in Leshan city of Sichuan province contains high contents of effective components， followed by Qujing and Wenshan cities of Yunnan province. Zingiberis Rhizoma Recens planted in Shandong and other places is mostly edible.

Analysis on Indications and Medication Regularities of Famous Classical Formula Wuzhuyu Tang Based on Modern Clinical Literature / 中国实验方剂学杂志

Wuzhuyu Tang （WZYT）， a classical traditional Chinese medicine （TCM） formula， was first recorded in Treatise on Cold Damage by the prestigious physician ZHANG Zhong-jing in the Eastern Han Dynasty， and included in Catalogue of Ancient Classical Formulas （First Batch）. WZYT is composed of four medcinals： Euodiae Fructus， Ginseng Radix et Rhizoma， Zingiberis Rhizoma Recens， and Jujubae Fructus， which has the effects of warming the middle and tonifying deficiency， dissipating cold and descending adverse qi. Based on data mining， this paper analyzes the modern clinical research literature on WZYT， which is expected to lay a basis for clarifying the "disease-symptom-prescription-medicinal" relationship of WZYT， as well as its modern clinical indications and clinical dosage. With the search term "Wuzhuyu Tang"， 253 complete medical records were retrieved from CNKI， VIP， and Wanfang （2000-2020）， involving 292 TCM prescriptions. Microsoft Excel 2019 was employed to establish the medical record database and the main clinical indications were statistically analyzed. Moreover， the syndromes， incidence law， and medication regularity were summed up. The result indicates WZYT has a wide range of modern clinical applications， mainly including the treatment of headache， dizziness， vomiting， diarrhea， dysmenorrhea， morning sickness， and other diseases， and the "deficiency， cold， and reversal" symptoms， mainly manifested as nausea and vomiting， headache， fatigue and lack of strength， reversal cold of hands and feet， poor appetite， gloomy complexion， dizziness， aversion to wind and cold， abdominal pain， sloppy diarrhea， abdominal distension， poor sleep， pale tongue， white coating， and deep thread wiry pulse. These diseases and symptoms mainly occur in women and middle-aged people. In most cases， medicinals are added or subtracted according to the symptoms， and the common doses of Euodiae Fructus， Zingiberis Rhizoma Recens， Codonopsis Radix or Ginseng Radix et Rhizoma， and Jujubae Fructus in the original prescription are 6-15 g， 10-15 g， 10-15 g or 9-12 g， and 4-6， respectively. The added medicinals are in the descending order of Pinelliae Rhizoma， Atractylodis Macrocephalae Rhizoma， Poria， Glycyrrhize Radix et Rhizoma， and Chuanxiong Rhizoma according to the frequency.

Views on Key Technical Problems in Research and Development of Famous Classical Formulas / 中国实验方剂学杂志

In this paper， the key technical problems in the research and development of famous classical formulas are analyzed. Firstly， the puzzled problem for a long-time， which is conversion relationship from medicinal metrology of Han dynasty （HD） to that of modern （gram，g）， is comprehensively expounded that one Liang （两） of HD=3 g is more appropriate. Secondly， the model and principles of quality consistency evaluation are given for the transformation from the quality of authoritative basic sample prepared by casserole （ABS-C） to the quality consistency in Laboratory process， pilot-scale process and industrial production. The consistency evaluation model is ξABS-X=K1（Q1ABS-X/Q1ABS-C）+K2（Q2ABS-X/Q2ABS-C）+……+Ki（QiABS-X/QiABS-C）=∑Ki（QiABS-X/QiABS-C）（i=1，2，3……n）. In the formula， ABS-X means laboratory reference sample ABS-C （ABS-L）， pilot-scale ABS-C （ABS-mP） or industrial production ABS-C （ABS-P）， ξABS-X means the quality consistency rate or similarity degree of ABS-L， ABS-mP and ABS-P processes with ABS-C， Ki means the weight of each quality evaluation index （i）， QiABS-X is the data of i in ABS-L， ABS-mP， ABS-P samples， and QiABS-C is the data （or mean） of i in ABS-C sample. Thirdly， in order to control the quality of the herbal medicines whose active ingredients were unknown， their chemical constituents should be studied deeply， and if necessary， the bioassay research should be carried out according to the main efficacy or indication of famous classical formulas. Finally， for the special processing of some herbal medicines， it is difficult to formulate the processing method， technology and standard of prepared slices. It is suggested that the scientific connotation and historical evolution of the special processing method should be thoroughly sorted out， and its technological characteristics are summarized， the modern processing technology and production processes are simulated， and then the corresponding processing methods and quality standards are formulated.

Guiding significance of "disease-syndrome-symptom" mode in FU Qing-zhu's Obstetrics and Gynecology (FU Qing-zhu Nyu Ke) for dealing with ovulation disorder infertility caused by hyperprolactinemia / 中国中药杂志

This paper discussed the guiding significance of "disease-syndrome-symptom" mode in FU Qing-zhu's Obstetrics and Gynecology(FU Qing-zhu Nyu Ke) for dealing with ovulation disorder infertility caused by hyperprolactinemia(HPRL). FU Qing-zhu's Obstetrics and Gynecology(FU Qing-zhu Nyu Ke) concentrates on the disease entities, main symptoms, pathogenesis, and syndrome differentiation, based on which the prescriptions are prescribed. This reflects the "disease-syndrome-symptom" mode, with the core lying in the "combination of disease with syndrome". The contained Discussion on Menstruation Regulation(Tiao Jing Pian) and Discussion on Getting Pregnant(Zhong Zi Pian) have important reference significance for later doctors in the diagnosis and treatment of inferti-lity, and many prescriptions are still in use due to good effects. It is believed in traditional Chinese medicine(TCM) that HPRL results from kidney deficiency and liver depression, among which kidney deficiency is the main cause. Liver depression accelerates the onset of HPRL, so the kidney-tonifying and liver-soothing herbs were mainly selected. The "disease-syndrome-symptom" mode in FU Qing-zhu's Obstetrics and Gynecology(FU Qing-zhu Nyu Ke) sheds enlightenment on the diagnosis and treatment of ovulation infertility caused by HPRL, in that it is not confined to disease entity and syndrome type. The integration of "disease-syndrome-symptom" highlights the main complaint of patients and emphasizes the main pathogenesis, thus giving full play to the overall advantage of syndrome differentiation. For multiple diseases in FU Qing-zhu's Obstetrics and Gynecology(FU Qing-zhu Nyu Ke) such as infertility due to liver depression, infertility due to obesity, delayed menstruation, and irregular menstruation, although the typical lactation symptom of HPRL is not mentioned, the medication can still be determined according to the chief complaint, syndrome type, and symptoms and signs, making up for the defects of excessive reliance on serum biochemical indicators in modern Chinese medicine. We should learn its diagnosis and treatment thoughts of paying attention to liver, spleen, kidney, and heart, holism, and strengthening body resistance to eliminate pathogenic factors.

Research progress of tannins in traditional Chinese medicines in recent ten years / 中国中药杂志

In this paper, the newly isolated tannins were sorted after a review of the literature concerning tannins in recent 10 years, and their research progress was summarized in terms of extraction, isolation, pharmacological activity and metabolism. Hydrolysable tannins and condensed tannins are the main structural types. Modern research shows that tannins have many pharmacological effects, such as bacteriostasis, antioxidation, antitumor, antivirus and blood glucose reduction, and have broad development prospects. They are usually extracted by water, ethanol and acetone and isolated and purified by macroporous resin and gel column chromatography. The packings commonly adopted for the column chromatography mainly included Sephadex LH-20, Diaion HP-20, MCI-gel CHP-20 and Toyopearl HW-40. Modern analytical techniques such as nuclear magnetic resonance spectroscopy(NMR), fast atom bombardment mass spectrometry(FAB-MS) and circular dichroism(CD) are generally used for the structural identification of tannins. Howe-ver, their isolation, purification and structural identification are still challenging. It is necessary to use a variety of high-throughput screening methods to explore their pharmacological activities and to explore the material basis responsible for their functions through experiments in vivo.

Content determination and transferring rules of seven components in Angelicae Sinensis Radix and its processed products / 中国中药杂志

A HPLC method was established for simultaneous determination of two organic acids(chlorogenic acid and ferulic acid) and five phthalides(senkyunolide I, senkyunolide H, senkyunolide A, ligustilide, and butylidenephthalide) in Angelicae Sinensis Radix and its processed products to clarify the underlying material transferring rules. The analysis was performed on a Welch Ultimate C_8 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.085% phosphoric acid water(B) as the mobile phase in a gradient elution mode at the flow rate of 1.1 mL·min~(-1), the column temperature of 25 ℃, the detection wavelength of 280 nm, and the injection volume of 10 μL. Under these conditions, the content of the above-mentioned seven components was analyzed in 15 batches of Angelicae Sinensis Radix and its processed products, and the transfer rate of each compound was calculated. As a result, in the processed products, the average content of chlorogenic acid was slightly decreased and that of ferulic acid was equivalent to the medicinal materials. The content of senkyunolide I, senkyunolide H, senkyunolide A, and butylidenephthalide showed an increasing trend in the processed products as compared with the medicinal materials. The mass fraction of ligustilide in the medicinal materials was above 0.7%(0.94% on average), meeting the requirement of 0.6% in the Hong Kong Chinese Materia Medica Standards, but was 0.47% on average in the processed products, which was decreased by 50% approximately. Further investigation showed that the content of ligustilide in freshly made processed products of Angelicae Sinensis Radix did not change significantly compared with that in the medicinal materials, indicating that the loss of ligustilide in the processed products mainly occurred in the storage. Therefore, Angelicae Sinensis Radix is suitable for storing in the form of medicinal materials and the freshly made processed products should be used except for special cases. Additionally, it is recommended to control the content of volatile oils or ligustilide in medicinal materials and processed products of Angelicae Sinensis Radix to ensure its effectiveness in clinical medication.

Research progress on Polygalae Radix / 中国中药杂志

Polygalae Radix has long been used in China for calming the mind, promoting intelligence, communicating the heart and kidney, eliminating phlegm, and reducing swelling. At present, it is used to treat amnesia, insomnia, and malaise. Modern research has revealed that Polygalae Radix mainly contains triterpenoid saponins, xanthone, oligosaccharide esters, etc., with the activities of improving memory, resisting dementia, protecting the brain, relieving cough, and removing phlegm, as well as sedation and hypnosis. The present study reviews the research progress on chemical composition, pharmacological action, quality control, and metabolism of Polygalae Radix in the past 30 years, to provide a theoretical basis for further research and development.

HPLC fingerprint of Tibetan medicinal plant ″Bangga″ and chemical components analysis of its two original plants by LC-MS/MS / 中国中药杂志

To obtain the chemical profile of Tibetan medicinal plant ″Bangga″, the present study established the HPLC fingerprint of ″Bangga″ and inferred common chemical constituents of its two original plants, Aconitum tanguticum and A. naviculare by LC-MS. The HPLC analysis was performed on a Kromasil 100 C_8 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.1% formic acid in water(B) as mobile phase in a gradient elution mode. Besides, the flow rate was set at 1 mL·min~(-1) and the column temperature was 35 ℃. The detection wavelength was set at 255 nm and the injection volume was 10 μL. Seventeen batches of ″Bangga″ samples were analyzed and the HPLC fingerprint was established under the above conditions. Similarity evaluation was performed using Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012). As a result, 16 common peaks were selec-ted and the similarity values of 17 batches of ″Bangga″ were in the range of 0.702-0.966. Furthermore, one batch of A. tanguticum and one batch of A. naviculare were analyzed by LC-MS/MS and 74 common compounds were inferred, including 10 phenolic acids, 26 flavonoids, and 38 alkaloids. The established method, with good separation and strong specificity, is simple and feasible, and can be used for the quality control of ″Bangga″ and identification of its two original plants. A. tanguticum and A. naviculare are similar in chemical composition and component content, but are quite different in the content of flavonoids.

Pharmacokinetics of the main components differing between Ligustri Lucidi Fructus and its wine-steamed product / 药学学报

A quantitative analysis method for ten principal components (phenylethanol, iridoids and triterpenes) of raw Ligustri Lucidi Fructus and its wine-steamed product was developed using liquid chromatography tandem triple quadrupole mass spectrometry (LC-QQQ-MS) to study their pharmacokinetic behavior in vivo. The results of methodological investigation were in accord with the criteria of biological analysis. After a single administration to rats of the water extracts of Ligustri Lucidi Fructus and its wine-steamed product, the plasma concentration of each component at different time points was measured and the pharmacokinetic parameters were determined. The AUC0-24 h and Cmax of the phenylethanol components (salidroside, tyrosol, hydroxytyrosol) were the greatest, suggesting that these components are the main pharmacological substances of Ligustri Lucidi Fructus. In addition, the tmax values of the eight major components were even lower with administration of the wine-steamed product, suggesting that these components are rapidly absorbed. However, the tmax values of specnuezhenide and oleanolic acid were greater with administration of the wine-steamed product, indicating that these two components were more slowly absorbed. A secondary peak phenomenon of tyrosol and hydroxytyrosol were observed in two sample groups, whereas the secondary peak phenomenon of salidroside occurred only with the wine-steamed product. This result suggests that the effect of wine-steamed product could persist for a long period. Meanwhile, the relative bioavailability of specnuezhenide and oleanolic acid was greater than 100% with administration of the wine-steamed product, consistent with the Traditional Chinese Medicine theory of the wine-steamed product being more effective than the raw material. The results reveal the different pharmacokinetic parameters and relative bioavailability of each component of Ligustri Lucidi Fructus and its wine-steamed product, and also demonstrate the variation and correlation of various components in vivo and in vitro, providing an experimental basis for the selection of quality control indexes, mechanisms of processing and the metabolic rule in vivo of Ligustri Lucidi Fructus. These experiments were approved by the Ethics Committee of Institute of Basic Theory for Chinese Medicine, China Academy of Chinese Medicine Science.

Analysis of Chemical Constituents in Euodiae Fructus by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To analyze the chemical constituents in Euodiae Fructus by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. Method:The chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with acetonitrile （A）-0.1% formic acid aqueous solution （B） as mobile phase （0-3 min， 6%A； 3-4 min， 6%-10%A； 4-7 min， 10%-12%A； 7-8 min， 12%-14%A； 8-13 min， 14%-15%A； 13-15 min， 15%-20%A； 15-18 min， 20%-30%A； 18-21 min， 30%-49%A； 21-25 min， 49%-51%A； 25-27 min， 51%-73%A； 27-30 min， 73%-80%A； 30-31 min， 80%-100%A； 31-32 min， 100%A） for gradient elution. The column temperature was 35 ℃， and the flow rate was 0.4 mL·min^-1. Mass spectrometry was performed using an electrospray ionization and data were collected in positive and negative ion modes， and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 200. The chemical constituents in Euodiae Fructus were identified rapidly and comprehensively based on the accurate relative molecular mass and combined with literature data and reference substances. Result:A total of 92 chemical constituents were speculatively identified from the 70% methanol extract of Euodiae Fructus， including 39 alkaloids， 19 flavonoids， 12 limonoids， 20 phenolic acids and 2 organic acids. Among them， 26 compounds were confirmed by the reference substances. Conclusion:The compound types of Euodiae Fructus are multiple and quite different in polarity. The chemical compositions of Euodiae Fructus from different regions and species are similar. The established method is rapid and accurate， with which the chemical compositions of Euodiae Fructus have been identified comprehensively. Therefore， this study provides an experimental reference for further clarifying active and toxic constituents of Euodiae Fructus.

Discussion for Quality Control of Chinese Patent Drugs from Quality Difference Among Different Dosage Forms of Liuwei Dihuang Preparations / 中国实验方剂学杂志

Objective:Through comprehensive evaluation and analysis of the quality of Liuwei Dihuang （LWDH） preparations from different manufacturers and combining factors such as production technology， the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations. Method:Morroniside， loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A） -0.3% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-8%A； 5-20 min， 8%A； 20-35 min， 8%-20%A； 35-45 min， 20%-60%A； 45-55 min， 60%A）， the detection wavelength of paeonol was at 274 nm， and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms （big candied pills， water-honeyed pills， concentrated pills， hard capsules and soft capsules） from different manufacturers were analyzed. Combined these results with their actual production processes， the key-points of quality control in the whole production process were discussed. Result:The contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of <italic>Chinese Pharmacopoeia</italic>， however， the content limit of some dosage forms in the current standard was unreasonable. For example， although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills （15.00， 8.57 g， respectively）， they got exactly the same daily limits of the contents for both the quality markers. What′s more， these two formulations had the same process， so the differences between the process obviously could not be the reason of these differences. Conclusion:It is recommended that for the products with different dosage forms should have a similar content limits， if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards， and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.

Investigation on in Vitro Antioxidant Activity and Chemical Composition of Different Polar Parts of Extract of Rosa cymosa Roots / 中国实验方剂学杂志

Objective:To investigate the antioxidant activity and chemical composition of 75% ethanol extract of <italic>Rosa cymosa</italic> roots and its different polar parts. Method:The 75% ethanol extract of <italic>R. cymosa</italic> roots was divided into dichloromethane， ethyl acetate， <italic>n</italic>-butanol and water parts by organic solvent extraction. <italic>In vitro</italic> antioxidant activity of each fraction was determined by 1，1-diphenyl-2-picrylhydrazyl （DPPH） and 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） free radical scavenging assays， as well as ferric reducing antioxidant power （FRAP） test. The contents of total triterpenes， total phenols， total tannins and condensed tannins in each fraction were determined by spectrophotometry. SPSS 24.0 software was used to conduct Pearson correlation analysis between the antioxidant activity of each fraction and the content of the main components， and then the main active fraction and the main active components were determined. The chemical constituents of the active fraction was analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）， and the structures of the main chromatographic peaks were predicted. Result:Each fraction of <italic>R. cymosa</italic> roots had certain antioxidant activity， and there was a significant dose-effect relationship within a certain concentration range， but the antioxidant activity of different polar parts was different. In DPPH and ABTS free radical scavenging tests， the antioxidant activity of each fraction and vitamin C （VC， the positive drug） was ranked as ethyl acetate fraction>VC><italic>n</italic>-butanol fraction>ethanol extract>water fraction>dichloromethane fraction. In FRAP test， the activity of ethyl acetate fraction was weaker than that of VC， and the other order was unchanged. The contents of total triterpenes， total phenols， total tannins and condensed tannins in ethyl acetate fraction were 3.81%， 50.33%， 3.32%， and 39.79%， in <italic>n</italic>-butanol fraction were 0.88%， 41.42%， 2.25% and 23.55%， in ethanol extract were 2.90%， 41.95%， 3.43% and 20.14%， in water fraction were 0， 26.80%， 16.90% and 7.57%， and in dichloromethane fraction were 21.23%， 12.90%， 1.59%， and 6.17%， respectively. Correlation analysis results showed that the contents of total phenols and condensed tannins were positively correlated with the antioxidant activity， the contents of total triterpenes were negatively correlated with the antioxidant activity， and the correlation between total tannins and antioxidant activity was not obvious. A total of 26 compounds were identified from the ethyl acetate fraction by UPLC-Q-TOF-MS/MS， including 11 condensed tannins， 4 hydrolysable tannins， 6 triterpenes， 3 flavonoids， 1 benzoic acid derivative and 1 chlorogenic acid analogue. Conclusion:Ethyl acetate fraction is the main antioxidant active site of <italic>R. cymosa</italic> roots， and phenols mainly composed of condensed tannins are the main active components. The results can provide experimental basis for the development of natural antioxidants.

Anti-acetylcholinesterase Active Ingredients from Aconitum tanguticum by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To identify the anti-acetylcholinesterase active ingredients in <italic>Aconitum tanguticum</italic>， so as to lay the foundation for finding new anti-Alzheimer's disease （AD） drugs. Method:The anti-acetylcholinesterase active fractions of <italic>A. tanguticum</italic> were screened by the modified Ellman's method， and the chemical composition of the active fraction was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×50 mm， 1.7 μm） with acetonitrile （A）-0.4% ammonia aqueous solution （B） as mobile phase for gradient elution， and the column temperature was set at 30 ℃ with the flow rate of 0.4 mL·min^-1. Phase A of the dichloromethane fraction changed with time as follows：0-3 min， 5%A； 3-7 min， 5%-20%A； 7-11.5 min， 20%-33%A； 11.5-15.5 min， 33%-50%A； 15.5-20.5 min， 50%-80%A； 20.5-23 min， 80%-85%A； 23-25 min， 85%-95%A. Phase A of the <italic>n</italic>-butanol fraction changed with time as follows：0-2 min， 5%A； 2-8 min， 5%-20%A； 8-11 min， 20%-33%A； 11-15 min， 33%-95%A. Mass spectrometry was performed on electrospray ionization， data were collected in positive ion mode， and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 500. Result:Both the dichloromethane and <italic>n</italic>-butanol fractions had a certain inhibitory effect on acetylcholinesterase， their half inhibitory concentration （IC₅₀） values were （64±4.4） mg·L^-1 and （85.7±3.8） mg·L^-1， respectively. By UPLC-Q-TOF-MS/MS analysis， a total of 21 alkaloids were identified from the dichloromethane fraction， and 11 alkaloids were identified from <italic>n</italic>-butanol fraction. Guan-fu base Ⅰ， found in both fractions， was first discovered in <italic>A. tanguticum</italic>. Conclusion:Diterpene alkaloids are the main anti-acetylcholinesterase substances of <italic>A. tanguticum</italic>， which is worth further exploration.

Quality standard of Perillae Folium based on multicomponent determination with HPLC method / 中国中药杂志

This research established the HPLC methods for the determination of perillaketone, perillaldehyde, caffeic acid, scutellarin, and rosmarinic acid in 33 batches of Perillae Folium. Kromasil C_(18)(4.6 × 250 mm, 5 μm) chromatographic column was used, and the mobile phase for determination of the perillaketone and perillaldehyde was methanol-water(55∶45) solution, at a flow rate of 1.0 mL·min~(-1), with the column temperature at 30 ℃. The mobile phase for the determination of caffeic acid, scutellarin and rosmarinic acid was methanol(A)-0.2% phosphoric acid aqueous solution(B) with gradient elution(0-20 min, 25%-30% A; 20-60 min, 30%-43% A). The flow rate was 1.0 mL·min~(-1) and the column temperature was set at 30 ℃. The results showed that the established method can achieve good separation of the five components in samples, with a good linear relationship and high accuracy, indicating that the methods can be used for the determination of Perillae Folium. The results showed that all samples contained five components. And the content of rosmarinic acid(0.04%-1.57%) > scutellarin(0.03%-0.77%) > perillaldehyde(0.02%-0.66%) > perillaketone(0.03%-0.30%) > caffeic acid(0.006%-0.07%). Thirty-three Batches of Perillae Folium can be grouped into 5 categories. There are certain content rules and region specificities under different clusters. Perillaketone, perillaldehyde, and rosmarinic acid can be used as the main markers to evaluate the quality of Perillae Folium.

Discussion on rationality of Yinpian commodity grades--Sophora flavescentis as an example / 中国中药杂志

In recent years, the establishment of the commercial grade of Yinpian [traditional Chinese medicine(TCM) pieces for decoction] in the TCM industry has been hotly disputed. In this article, Sophorae Flavescentis Radix(SFR) was selected as a representative example to investigated. Through systematic comparison and analysis, the different grades of SFR slices were traced, verified and evaluated. According to the current published local grade standards of SFR slices, the results showed that the first-class of SFR slices were mostly derived from the wild medicinal materials, the second-class were mostly originated from the cultivated materials in 3-4 years, and the third-class products were from a small number of lateral roots and short-growing years or harsh habitat of wild medicinal materials. On the basis of identifying the sources of different grades of SFR slices, the contents of the active components, including matrine, oxymatrine, oxysophocarpine, sophoridine, N-methyl-cytisine, sophocarpine, were quantitatively determined in typical samples, it was found that the grades were inversely proportional to the contents of active ingredients. In order to ensure the universality of the conclusion, the contents of different grades of commercial SFR slices were determined, and the conclusion was verified as "the commercial grades of SFR slices were inversely linked to their contents of active ingredients". This phenomenon is common in the determination of the commercial grade of Yinpian of radix and rhizome. Therefore, we propose that the method or standard of the commercial grade of Yinpian of radix and rhizome based on the size of Yinpian maybe not proper. Whether and how to classify Yinpian commercial grade is not only a multi-disciplinary issue, especially in combination with clinical efficacy, but also a big problem need to consider the production, commercial circulation and other processes link of quality risk and quality assurance, and should be treated with great caution.

Determination of β-nicotinamide mononucleotide and nicotinamide adenine dinucleotide in Dendrobium officinale and congeneric species by UPLC-MS/MS / 中国中药杂志

As anti-aging ingredients, β-nicotinamide mononucleotide(NMN) and nicotinamide adenine dinucleotide(NAD~+) have attracted worldwide attention in recent years. After oral administration, NMN can be converted into NAD~+ in vivo and the latter is the actual ingredient which exerts anti-aging effect. In order to explore the "rejuvenating and anti-aging" effect of Dendrobium officinale, which was firstly recorded in Shennong's Herbal Classic of Materia Medica, this study established the quantitative method of UPLC-MS/MS for simultaneous determination of NMN and NAD~+ in D. officinale and the congeneric species for the first time, and 34 batches of samples were detected. UPLC conditions are as follows: ACQUITY UPLC HSS T3 column(2.1 mm × 100 mm, 1.8 μm), gradient elution with acetonitrile-0.1% formic acid in water at the flow rate of 0.3 mL·min~(-1), and column temperature of 40 ℃. MS conditions were scanned electrospray ionization source and multiple reaction monitoring mode. The method was verified by systematic methodology. The mean recoveries of NMN and NAD~+ were 77.58% and 80.70%, respectively, with RSD of 3.6% and 4.3%, separately. All results showed that the content of NMN was higher in D. officinale than in the other congeneric species. Particularly, the content in fresh D. officinale stems was as high as 0.931 9 μg·g~(-1). NAD~+ was only found in D. officinale and the content was three times higher than that of NMN. This may be the reason that D. officinale topped the "nine famous anti-aging herbs". In addition, processing method influences the content of NMN and NAD~+ in Dendrobium. Specifically, the content of NMN and NAD~+ was in the order of fresh Dendrobium stems > dried Dendrobium stem segments > spiral or spring-like dried Dendrobium stems.

Rapid discovery of tyrosinase inhibitors from Sophora flavescens:significance for comprehensive utilization of its non-medicinal resources / 中国中药杂志

Sophorae Flavescentis Radix,derived from the root of Sophora flavescens in the Leguminosae family,has been widely used in the medicine,agriculture,animal husbandry,and daily chemical industry. A pharmacophore model-based method for rapid discovery of tyrosinase inhibitors( TIs) from S. flavescens was established by molecular docking under Lipinski rules,and verified by enzyme assays. Briefly,the chemical constituent database of S. flavescens( CDSF) was established based on the previous papers. Theoptimal pharmacophore model( OPM) was constructed by DS 2019 on the basis of known active TIs. Eighty-three hits predominated by flavonoids having higher fitting scores with OPM than the positive control were screened out,and subjected to molecular docking based on the three-dimensional structure of tyrosinase crystal protein. The potential TIs such as kurarinone and nor-kurarinone were rapidly discovered from the compounds with higher docking scores than the positive control under the Lipinski rules. The results were verified by the in vitro enzyme assays. The inhibition activities of tyrosinase from non-medicinal parts of S. flavescens were also tested to explore the relationship between the inhibition activity and chemical compositions. This study is expected to provide data support for the comprehensive application and development of S. flavescens and also a new method for the rapid discovery of active substances or functional constituents in the complex systems.

Qualitative and quantitative analysis of nucleosides and nucleobases in Phylloporia ribis by UPLC-Q-TOF-MS and HPLC / 中国中药杂志

The dried fruit body of Phylloporia ribis(Hymenochaetaceae), which prefers to live on the stumps of Lonicera japonica(Caprifoliaceae), has a variety of activities, whereas its pharmacodynamic material basis is not completely clear and there are few reports on its quality control and evaluation. In this study, an UPLC-Q-TOF-MS method was used to analyze the nucleosides and nucleobases in P. ribis and a HPLC method was established for simultaneous determination of 10 nucleosides and nucleobases. MS and MS/MS data were acquired in positive ion mode. Based on the data comparison of the sample and the reference substance, the literature data and the compound databases of ChemSpider and PubChem, 18 nucleosides and nucleobases were identified qualitatively from the water extract of P. ribis for the first time. After optimization, the HPLC was performed using a Welch Ultimate AQ C_(18) column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile and water as mobile phase, the flow rate of 1.0 mL·min~(-1), the detection wavelength of 260 nm, and the column temperature of 30 ℃. Through the investigation of the extraction method, solvent and time, it was determined that the test solution should be obtained by cold water extraction for 18 h. At the present HPLC conditions, 10 components of uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine and thymidine could be well separated(R > 1.5) and showed good linearity(r > 0.999 9) in the concentration ranges of 0.247-24.7, 0.283-28.3, 0.273-27.3, 0.256-25.6, 0.257-25.7, 0.318-31.8, 0.245-24.5, 0.267-26.7, 0.250-25.0 and 0.267-26.7 mg·L~(-1), respectively. The average reco-veries of 10 components were 95.78%-104.5%, and the RSDs were 2.2%-5.2%(n=6). The contents of 10 nucleosides and nucleobases in different samples of P. ribis varied greatly, which were 0.021-0.122, 0.004-0.029, 0.014-0.226, 0.009-0.442, 0.003-0.014, 0.002-0.146, 0.007-0.098, 0-0.054, 0.005-0.069, 0.004-0.081 and 0.072-1.28 mg·g~(-1) for uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine, thymidine and total 10 components, respectively. These results demonstrated that the components had significant differences in the internal quality, and good quality control was needed to ensure the medical efficacy. This study provides a scientific basis for the discovery of pharmacodynamic ingredients, quality control and evaluation of P. ribis.

Examples of Professor MA Kun's treatment of infertility caused by hyperprolactinemia with kidney deficiency and blood stasis / 中国中药杂志

Hyperprolactinemia(HPRL) is one of the diseases leading to anovulatory infertility, which is a refractory gynecological disease and seriously affects female reproductive function. Professor MA Kun has summarized his experience in clinical and scientific studies for many years. And believes that kidney deficiency is the pathogenesis of HPRL and blood stasis is the dominant pathological manifestation of HPRL and can promote the progress of the disease. In view of this, Professor MA Kun took the therapy of kidney-tonifying and blood-activating as the principle for treating anovulatory infertility caused by HPRL, with soothing the liver and promoting Qi as adjuvant therapies. She has also summarized and refined the prescriptions for tonifying kidney and inducing ovulation, which have a remarkable clinical efficacy.

Qualitative and quantitative analysis on triterpenoids in Ligustri Lucidi Fructus / 中国中药杂志

Triterpenoids are one of the most active constituents in Ligustri Lucidi Fructus, but only oleanolic acid has been mostly studied. In recent years, a growing number of studies have shown that other triterpenes from Ligustri Lucidi Fructus also have various biological activities, so it is necessary to build up a detailed profile of the triterpenoids in Ligustri Lucidi Fructus. The chromatographic separation was performed on a C_(18) column(4.6 mm×250 mm, 5 μm) with mobile phase of methanol-acetonitrile-0.2% formic acid for gradient elution. The detection wavelength was set at 210 nm, with a flow rate of 0.5 mL·min~(-1), and the column temperature of 25 ℃. The HPLC fingerprint of triterpenoids in Ligustri Lucidi Fructus was built by testing 21 batches of samples from different sources. The structures of the total 15 common chromatographic peaks were elucidated with UHPLC-ESI-Orbitrap-MS/MS technique and six of them were identified as tormentic acid, pomolic acid, maslinic acid, botulin, oleanolic acid and ursolic acid by comparison to the reference substances. Under the same chromatographic condition, four main triterpenes(podocarboxylic acid, hawthorn acid, oleanolic acid and ursolic acid) were quantified and the results of system adaptability and methodology investigation all met the requirements of content determination. Meanwhile, with oleanolic acid(A) as the internal reference substance, quantitative analysis of multi-components by single marker(QAMS) method was used to analyze the above four components. The relative correction factor of oleanolic acid(B), hawthorn acid(C) and ursolic acid(D) to oleanolic acid was f_(B/A)=1.12, f_(C/A)=1.02 and f_(D/A)=0.88, respectively, and the relative retention values of these three to oleanolic acid was RRV_(B/A)=0.46, RRV_(C/A)=0.70 and RRV_(D/A)=1.03, respectively. The contents determined by two methods were similar. In conclusion, the method built in this paper is proved to be simple, reliable and specific for the simultaneous qualitative and quantitative analysis of the triterpenoids in Ligustri Lucidi Fructus, which can lay foundation for further assays of the triterpenoids in Ligustri Lucidi Fructus and the relative products.