Citri Reticulatae Pericarpium Research Articles

Effect of the Phytochemical Agents against the SARS-CoV and Some of them Selected for Application to COVID-19: A Mini-Review.

The aim of the present review is to provide basic knowledge regarding the treatment of Coronavirus via medicinal plants. Coronavirus (COVID-19, SARS-CoV, and MERS-CoV) as a viral pneumonia causative agent, has infected thousands of people in China and worldwide. Currently, there is no specific medicine or vaccine available that can treat or prevent this virus and this has posed a severe threat to human health; therefore, there is an urgent need to develop a novel drug or anticoronavirus vaccine. However, natural compounds to treat coronaviruses are the most effective alternative and complementary therapies due to their diverse range of biological and therapeutic properties. We performed an open-ended, English restricted search of Scopus database, Web of Science, and Pubmed for all available literature from Jan-March, 2020, using terms related to phytochemical compounds, medicinal plants and coronavirus. The view on anti-coronavirus (anti-CoV) activity in the plant-derived phytochemicals and medicinal plants gives a strong base to develop a novel treatment employing these compounds for coronavirus. Various phytochemicals and medicinal plant extracts have been revised and are considered as potential anti-CoV agents for effective control of the virus and future drug development. Herein, we discuss some important plants (Scutellaria baicalensis, Psorothamnus arborescens, Glycyrrhiza radix, Glycyrrhiza uralensis, Lycoris radiate, Phyllanthus emblica, Camellia sinensis, Hyptis atrorubens Poit, Fraxinus sieboldiana, Erigeron breviscapus, Citri Reticulatae Pericarpium, Amaranthus tricolor, Phaseolus vulgaris, Rheum palmatum, Curcuma longa and Myrica cerifera) that have emerged to have broad-spectrum antiviral activity. Nigella sativa has potent anti-SARS-CoV activity and it might be a useful source for developing novel antiviral therapies for coronavirus.

Analysis of medication regularity and pharmacodynamic characteristics of traditional Chinese medicine treatment in 444 severe cases of COVID-19

The coronavirus disease 2019(COVID-19) is developing rapidly and posing great threat to public health. There is no effective intervention for the severe patients, and their prognosis is poor. It is worth noting that in the fight against COVID-19, China has always put equal emphasis on both Chinese and Western medicine. Traditional Chinese medicine has played an important role in the whole process. It is of great significance to discuss the rules and characteristics of the prescription of traditional Chinese medicine in the treatment of COVID-19. In this study, information was collected from 444 severe COVID-19 patients who were admitted to a hospital designated to treat patients with severe COVID-19 in Wuhan before March 20, 2020. We collected traditional Chinese medicine prescriptions for patients with severe COVID-19, referred to Chinese Pharmacopoeia to standardize the names of traditional Chinese medicine, and extract the property, flavor and channel tropism of traditional Chinese medicines to analyze the rules of the prescriptions. IBM SPSS Modeler 18.0 software was used to conduct correlation analysis of traditional Chinese medicine. Effective traditional Chinese medicines against COVID-19 was identified by the TCMATCOV platform. In the end, 1 532 effective prescriptions were included. Among them, the high-frequency drugs are Poria, Astragali Radix, Pogostemonis Herba, Armeniacae Semen Amarum, Atractylodis Macrocephalae Rhizoma, Pinelliae Rhizoma, Glycyrrhizae Radix et Rhizoma, Magnoliae Officinalis Cortex, Ephedrae Herba, Cinna-momi Ramulus. Most of the drugs have the following functions: resolving dampness, replenishing deficiency, resolving phlegm, cough, and asthma. The core combinations are Pogostemonis Herba-Poria, Astragali Radix-Pogostemonis Herba-Poria, Amomi Fructus-Poria, Amomi Fructus-Pogostemonis Herba, Amomi Fructus-Astragali Radix. The majority of the medicines are with cold and warm properties, and the proportions are 41.03% and 38.46%, respectively. The medicinal flavors are mainly concentrated in sweet and bitter, and the proportions are 34.71% and 30.58%, respectively. The meridian of the drug is more into the lung, stomach and spleen, with lung accounting for 22.87%. From the analysis of high-frequency drugs to the core combinations, one can see that the main treatment principle for severe COVID-19 is to remove internal and external dampness, protect the spleen and stomach, remove evil energy, and support righteousness. TCMATCOV platform was used to calculate the network disturbances of the high-frequency drugs. It was found that the traditional Chinese medicine with a high disturbance score accounted for a high proportion of the classic anti-COVID-19 prescriptions used by clinicians. Among them, the drugs with top scores are Ephedrae Herba, Citri Reticulatae Pericarpium, Eupatorii Herba, Platycodonis Radix, Cinnamomi Ramulus, Astragali Radix, Magnoliae Officinalis Cortex, Atractylodis Macrocephalae Rhizoma, Pogostemonis Herba, Scutellariae Radix. After a further exploration of the action targets, it was showed that disease-specific factor TNF was the target of the above ten drugs, and traditional Chinese medicine can exert anti-inflammatory and immune-modulating effects.

Network pharmacology research on high frequency use of Pericarpium Citri Reticulatae and Aucklandiae Radix herb pair in treatment of constipation with aromatic traditional Chinese medicine based on data mining

Based on data mining and through the method of network pharmacology, we analyzed the mechanism of high-frequency use of herb pair in the treatment of constipation with aromatic traditional Chinese medicine in this study. Through data mining, aromatic traditional Chinese medicine was obtained for the treatment of constipation and Pericarpium Citri Reticulatae and Aucklandiae Radix herb pair was used as the research object. The volatile oil from Pericarpium Citri Reticulatae and Aucklandiae Radix was extracted by steam distillation, and the chemical compositions of the volatile oil were detected by gas chromatography-mass spectrometry(GC-MS). The targets of volatile oil from Pericarpium Citri Reticulatae and Aucklandiae Radix were searched by PubChem, TCMSP, STITCH and Swiss Target Prediction databases. The targets of constipation were predicted and screened in OMIM, Genecards-Search Resuits and TTD databases. The obtained targets were introduced into Cytoscape 3.7.1 to construct protein-protein interaction(PPI) network diagram for GO and KEGG pathway enrichment analysis by using R language. The network diagram of "component-target-pathway" was constructed according to the results of KEGG enrichment. Discovery Studio 2.5 software was used to verify the molecular docking between the components and the targets. Among them, the most frequently used pair of aromatic traditional Chinese medicine in the treatment of constipation was Pericarpium Citri Reticulatae and Aucklandiae Radix. A total of 33 compounds were detected by GC-MS, and a total of 180 common action targets of Pericarpium Citri Reticulatae and Aucklandiae Radix on volatile oil in the treatment of constipation were predicted. The key targets included CYP19 A1, PPARA, PGR, ACHE, SLC6 A2 and so on. GO enrichment analysis showed that the activities of Pericarpium Citri Reticulatae and Aucklandiae Radix on volatile oil were mainly involved in the biological processes such as circulatory system, blood circulation, and steroid hormone binding. In KEGG enrichment pathway, neuroactive ligand-receptor interaction, endocrine resistance, Ca~(2+) signal pathway and IL-17 signaling pathway showed significant effect on constipation. The results of molecular docking showed that PGR, the target protein related to the treatment of constipation, had a good binding with gamma-linolenic acid, dihydro-alpha-ionone, alpha-eudesmol, caryophyllene oxide and beta-ionone. The results show that by using data mining technology and network pharmacology, it is revealed that the active components of Pericarpium Citri Reticulatae and Aucklandiae volatile oil in high frequency use of aromatic traditional Chinese medicine can be used totreat constipation mainly through CYP19 A1, PPARA, PGR, ACHE, SLC6 A2 and other targets, providing a new idea and method for the further study of aromatic traditional Chinese medicine in the treatment of constipation.

Prescription and medication regularity of traditional Chinese medicine for treating child pneumonia based on data mining

To analyze and summarize formulae of traditional Chinese medicine(TCM) in the treatment of pediatric pneumonia in CNKI based on the data mining method. CNKI was retrieved for literatures of TCM treatment of pneumonia in children in recent 20 years, which was taken the data source. After screening, a prescription database was established. Frequency analysis, association rules Apriori algorithm and complex system entropy clustering analysis methods integrated in TCMISS(V2.5) were applied in data analysis and mining. Core drugs and their properties and flavors, medication modes and new prescriptions were summarized and studied. A total of 408 prescriptions were screened out, involving 218 drugs, among which Chinese herbal medicines with the highest use frequency included Gancao(Glycyrrhizae Radix et Rhizoma), Xingren(Armeniacae Semen Amarum), Huangqin(Scutellariae Radix), Mahuang(Ephedrae Herba), Jiegeng(Platycodonis Radix), Banxia(Pinelliae Rhizoma), Sangbaipi(Mori Cortex), Chenpi(Citri Reticulatae Pericarpium), Fuling(Poria), Jinyinhua(Japonicae Lonicerae Flos). Flavors were mainly bitter, sweet and pungent; and the main medicinal property was cold. Most of these drugs entered lung meridian(LU), stomach meridian(ST) and spleen meridian(SP); and the association rules between drugs were established. Totally 6 new prescriptions were obtained through entropy hierarchical cluster analysis. Our results objectively presented, at present, TCM focuses on phlegm and heat in treating children's pneumonia, which targets at lung and also treats spleen and stomach. The main therapies aim to clear heat and resolve phlegm, ventilate lung and relieve cough and dyspnea, release exterior and dissipate cold, and relieve cough and eliminate phlegm. The results of statistical analysis are basically consistent with the consensus of experts, which can provide theoretical basis for clinical medication and drug development.

Comparison of the content of flavonoids, total phenols, and carotenoids and antioxidant activity in guang Citri reticulatae pericarpium during the aging time

Background: Guang Citri reticulatae Pericarpium (GCRP) is a traditional Chinese medicine, widely used in respiratory and digestive diseases. GCRP must be aged by stored for several years before it can be used as medicine. In the aging process, content of composition, antioxidant activity, and color of GCRP have changed. Objectives: The objective of this study was to compare the content of flavonoids, total phenols, and carotenoids and antioxidant activity in GCRP during the aging time and analyze the correlations of the data. Materials and Methods: Forty batches of GCRP in different aging years were used for qualitative and quantitative analysis of flavonoids, total phenols, and carotenoids by ultra-high-performance liquid chromatography (HPLC)–mass spectrometry, HPLC–diode array detector, and ultraviolet methods. 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and 1,1-diphenyl-2-trinitrophenylhydrazine methods were used to compare the antioxidant activity of GCRP. Colorimetric analysis method was used to determine the external color of GCRP objectively and quantitatively. SPSS software was used to analyze the correlation of all data. Results: With the increase of aging year, the content of three flavonoids and total phenols increased and the content of carotenoids decreased. The results showed that the antioxidant activity of ether extract increased significantly with aging year and methanol extract decreased. The correlation results showed that the content of carotenoids was more closely related to chromaticity values and antioxidant activity. Conclusion: The result provides more objective and detailed information to analyze the quality of GCRP during the aging time.

Identification and Quantification of “Quality Markers” in Tongkang Tablet on Recurrent Respiratory Tract Infections (RRTIs) by UFLC-IT-TOF–MS/MS and HPLC–DAD

Recurrent respiratory tract infections (RRTIs) is one of the most common diseases in children and adolescents, especially for preschool children. Tongkang tablet (TKT), a traditional Chinese medical formula consisting of Astragali Radix (AR), Atractylodes Macrocephala (AM), Saposhnikovia Divaricata (SD), Rhizoma Dioscoreae (RD), Concha Ostreae (CO), and Pericarpium Citri Reticulatae (PCR), has been widely used to treat recurrent upper respiratory tract infections of children. To ensure the efficacy and safety of TKT, an integrative and reliable quality control method is urgently needed. The concept of quality marker (Q-marker) is highly helpful but the selection and quantitation of appropriate Q-markers remains challenging, especially for the complex components of Chinese medicinal products. The aim of this article was to identify and quantify Q-markers of TKT for the establishment of a quality control method. First, by a validated UFLC-IT-TOF–MS/MS method, a total of 40 compounds were unambiguously or tentatively identified, including 21 flavonoids, 10 chromones, 3 coumarins, 3 sesquiterpenes, 2 batatasins and 1 saponin. Among them, 10 compounds were further confirmed by reference standards. Then, 10 compounds, including calycosin-7-O-glucoside, prim-O-glucosylcimifugin, cimifugin, naringin, 4′-O-D-glucosyl-5-O-methylvisamminol, hesperidin, ononin, calycosin, sec-O-glucosyhamaudol and formononetin, were selected as Q-markers based on the knowledge of absorption characteristics and bio-activity with the reference literatures. Finally, based on the 10 Q-markers, the fingerprint chromatography was established by HPLC–DAD method, which proved accurate, sensitive and reliable. The presently developed method based on 10 Q-markers is expected to lay a good foundation for the quality control of TKT.

Effects of Chinese medicines on monacolin K production and related genes transcription of Monascus ruber in red mold rice fermentation.

Monacolin K (MK) is a secondary metabolite synthesized by polyketide synthases of Monascus spp. In this study, the combined supplementation of three medicines, including Citri Reticulatae Pericarpium (CRP), Fructus crataegi (FC), and Radix Angelicae Dahuricae (RAD), were mixed with nonglutinous rice and were optimized by response surface methodology to enhance the production of MK in fermented red mold rice (RMR). Under the optimum condition, MK production achieved 3.60 mg/g, which was 41.18% higher than RMR without medicines. The improved MK production was mainly caused by the up‐regulated transcription level of mokA, mokB, mokF, mokH, mokI, and mplaeA. Meanwhile, the inhibitory effect of Poria cocos (PC) on MK production (only 0.436 mg/g) was caused by significantly down‐regulated transcription of six tested genes. Therefore, this study is beneficial for better understanding of the possible mechanism of enhanced MK production by optimization of fermentation conditions.

A Rapid and Nondestructive Approach for the Classification of Different-Age Citri Reticulatae Pericarpium Using Portable Near Infrared Spectroscopy.

Citri Reticulatae Pericarpium (CRP), has been used in China for hundreds of years as a functional food and medicine. However, some short-age CRPs are disguised as long-age CRPs by unscrupulous businessmen in order to obtain higher profits. In this paper, a rapid and nondestructive method for the classification of different-age CRPs was established using portable near infrared spectroscopy (NIRS) in diffuse reflectance mode combination with appropriate chemometric methods. The spectra of outer skin and inner capsule of CRPs at different storage ages were obtained directly without destroying the samples. Principal component analysis (PCA) with single and combined spectral pretreatment methods was used for the classification of different-age CRPs. Furthermore, the data were pretreated with the PCA method, and Fisher linear discriminant analysis (FLD) with optimized pretreatment methods was discussed for improving the accuracy of classification. Data pretreatment methods can be used to eliminate the noise and background interference. The classification accuracy of inner capsule is better than that of outer skin data. Furthermore, the best results with 100% prediction accuracy can be obtained with FLD method, even without pretreatment.

Dual-QDs ratios fluorescent probe for sensitive and selective detection of silver ions contamination in real sample.

Silver ions, as a commonly used industrial heavy metal, tends to deposit in the body and induce many diseases. In this work, modified CdTe QDs with green and red emission were synthesized to assemble dual-QDs, which could be efficient and selective utilized for Ag+ determination through the electron transfer progress between surface functional group of dual-QDs and Ag+, and the aggregation of Ag+ on the surface of dual-QDs. Under the appropriate pH value and volume ratio, the interaction between the surface functional groups of assembled dual-QDs reduce the affinity of Hg2+ in this system. The fluorescent signal of dual-QDs simultaneously attenuation or enhancement in the same proportion remove the interference of Cu2+ and other metal ions. Therefore, this method can selectively detect Ag+ without any masking agents. The linear region of detection was from 0 to 800nmol/L (R2>0.998), and low of detection (LOD) was 7.7nmol/L, which could meet the corresponding standards of World Health Organization (WHO) and Environmental Protection Agency (EPA). This effective proposed dual-QDs ratios fluorescent probe has been applied to detect Ag+ in real environment water, tea and Citri Reticulatae Pericarpium (CRP) water.

Chemome profiling of Citri Reticulatae Pericarpium using UHPLC-IT-TOF-MS

Citri Reticulatae Pericarpium(CRP, Chinese name: Chenpi) is one of the most famous edible traditional Chinese medicines(TCMs). CRP was first recorded as top grade TCM in Shennong Bencao Jing attributing to the benefits such as regulating Qi, tonifying spleen, eliminating dampness and eliminating phlegm, and has been widely utilized for the treatments of abdominal fullness and distention, vomiting and diarrhea, as well as phlegm cough. CRP is also widely popular as spice in food industry. Because of the wide cultivation, a number of brands that exhibit extensive price range can be found in the market, resulting in a great challenge for grading. Herein, an attempt was made to in-depth chemome profiling for the sake of providing meaningful information of the universal quality control of CRP. A new core-shell column packed with adamantylethyl substituted silica gel particles was deployed for chromatographic separations and IT-TOF-MS that is advantageous at providing abundant high resolution molecular and fragment ions was employed for qualitative detection. A total of 62 components were observed and 61 ones were structurally annotated according to proposing mass fragmentation patterns, matching with reference compounds and relevant databases, and the chemical families included flavone, limonin, etc. In particular, ten compounds bearing 3-hydroxy-3-methylglutarate substitute were detected from CRP for the first time. Above all, the chemical profile of CRP was characterized and the findings are meaningful for the in-depth quality assessment and efficacy material clarification of CRP.

Ultra-small and biocompatible platinum nanoclusters with peroxidase-like activity for facile glucose detection in real samples

The highly sensitive glucose detection based on the peroxidase-like properties of nanoclusters has been gained great interest. In this work, Pericarpium Citri Reticulatae polysaccharide (PCRP) stabilized platinum nanoclusters (Pt-PCRP NCs) were prepared by a green method in which potassium tetrachloroplatinate and PCRP were simply mixed without addition of other agents. Platinum nanoclusters (Pt NCs) had ultra-small size of 1.26 ± 0.34 nm. The hydrodynamic size of Pt-PCRP NCs was 29.7 nm, and zeta potential of which was -12.0 mV. Pt-PCRP NCs showed high biocompatibility toward HeLa cells and red blood cells. In addition, Pt-PCRP NCs catalyzed the decomposition of H2O2 to produce •OH, which further oxidized colorless 3,3′5,5′-tetramethylbenzidine (TMB) to blue oxidized 3,3′,5,5′-tetramethylbenzidine (oxTMB), exhibiting peroxidase-like property. The kinetics followed the Michaelis-Menten equation. More importantly, the colorimetric method for glucose detection using Pt-PCRP NCs had high selectivity and low detection limit for 0.38 μM. The established method based on Pt-PCRP NCs was used to precisely detect glucose detection in human serum, saliva, and sweat. Taken together, the prepared ultra-small and biocompatible Pt-PCRP NCs have good potential glucose applications in clinical diagnosis in the future.

Construction and Chemical Profile on "Activity Fingerprint" of Citri Reticulatae Pericarpium from Different Cultivars Based on HPLC-UV, LC/MS-IT-TOF, and Principal Component Analysis.

Citri Reticulatae Pericarpium (CRP), known as Chenpi (CP) in Chinese, is a medicinal food for health and fitness. In order to find out the characteristic activity chemicals distinguishing various cultivars of CRP and provide a reference for effective development of citrus resources, an “activity fingerprint” of CRP from 21 different cultivars was established based on the evaluation of antitussive and expectorant activities. There were 18 common peaks in the HPLC fingerprint, of which 3 flavonoid glycosides and 14 polymethoxyflavonoids (PMFs) were identified by LC/MS-IT-TOF. Furthermore, five characteristic chemicals were determined and similarity calculation with principal component analysis (PCA) was combined together to compare the similarity and difference among various cultivars. The results showed that some other cultivars were also similar to C. reticulata “Chachi” except for C. reticulata “Tangerina” and C. reticulata “Dahongpao” recorded in Chinese Pharmacopoeia. Most importantly, the peels of C. reticulata “Shiyueju,” C. reticulata “Ponkan,” C. reticulata “Tribute,” and C. reticulata “Bayueju,” traditionally rarely used for medicinal food, were highly similar to that of C. reticulata “Chachi” and rich in bioactive flavonoids, which can be considered the effective medicinal resources of CRP.

A rapid protocol to distinguish between Citri Exocarpium Rubrum and Citri Reticulatae Pericarpium based on the characteristic fingerprint and UHPLC-Q-TOF MS methods.

Citri Exocarpium Rubrum (CER) and Citri Reticulatae Pericarpium (CRP) are used as common functional foods and traditional Chinese medicine (TCM). As different parts of the same fruit, CER and CRP have different effects in clinical applications. However, they are commonly confused due to the similarity of the chemical compounds and a lack of scientific method to distinguish them in the finished product. In this study, an ultra-high performance liquid chromatography-diode array detector (UHPLC-DAD) technique and an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) method were employed to generate the characteristic fingerprint under the optimum analytical conditions. Principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were applied to represent different chemical markers for CER and CRP. 44 potential markers including 15 polymethoxylated flavanones (PMFs), 5 flavone-C-glycosides, 6 flavanone-O-glycosides, 3 flavonoid-O-glycosides, 8 organic acids, 5 limonoids and 2 alkaloids were successfully identified by using UNIFI software. The heat map showed that there were significant differences in the CER and CRP samples. Furthermore, 12 potential markers were screened out by characteristic fingerprint and UHPLC-Q-TOF MS methods and were analyzed by quantitative analysis of multicomponents by a single marker (QAMS) method. Finally, a prediction model based on the discovered chemical markers was established for discrimination between CER and CRP. Using these markers can significantly distinguish the unknown processed products of CER and CRP. In conclusion, an effective way to quickly and easily distinguish CER and CRP was successfully established based on the characteristic fingerprint and UHPLC-Q-TOF MS. It could also be a new strategy for analysis of different processed products of the same plant source.

Development and validation of a supercritical fluid chromatography method for fast analysis of six flavonoids in Citri Reticulatae Pericarpium

The study is aimed to develop a rapid and efficient supercritical fluid chromatography coupled with diode-array detection (SFC-DAD) method for simultaneous quantification of six flavonoids in Citri Reticulatae Pericarpium (CRP). The chromatographic parameters including stationary phase, mobile phase composition, back pressure, temperature and flow rate, were systematically optimized with single-factor test. The results indicated that the six compounds were baseline separated on an Agilent Zorbax RX-SIL column using gradient elution within 10 min. The optimized mobile phase was constituted of carbon dioxide (CO2) and methanol. And the parameters were set as follows: backpressure, 95 bar; temperature, 45 °C; flow rate, 0.8 mL/min. The optimized method showed satisfactory retention and resolution for the analytes. The method was validated to demonstrate its selectivity, linearity, precision, accuracy and robustness. Thus, the verified SFC method was successfully applied to quantification of flavonoids present in CRP samples. The results indicated that SFC has the potential to become an excellent alternative for the analysis of flavonoids in herbal medicines, owning to its intrinsic features like high efficiency, separation speed and eco-friendly characteristics.

Rapid characterization of compounds in fupo ganmao granules by high-performance liquid chromatography tandem mass spectrometry

Fupo Ganmao Granules (FP) is a classic traditional Chinese medicine (TCM) formula, composed of 4 herbal medicines, Folium Hibisci Mutabilis, Magnolia officinalis, Pericarpium Citri Reticulatae, and Fructus arctii. It is regularly used in the treatment of common cold with wind-heat syndrome. The biologically active constituents in FP, especially those in trace amount, remain unclear. In this study, the components of FP were profiled and characterized using HPLC-ESI-MS2. A total of 58 compounds were identified according to their fragmentation features in IT-TOF-MS and IT-MS2, including 13 phenolic acids, 17 flavonoids, 21 lignans, 2 alkaloids, and 5 other compounds. Among them, the identities of 8 compounds were explicitly confirmed by comparing them with standard compounds, and their contents were determined. Our study provided a comprehensive understanding of the diverse chemical profile in FP, and the approach we developed is useful for the analysis of other TCM formulas.

The central bacterial community in Pericarpium Citri Reticulatae 'Chachiensis'.

The dried and aged pericarps of Citri Reticulatae are condiments and medicinal products in southeast and eastern Asia for hundreds of years, among which Pericarpium Citri Reticulatae 'Chachiensis' (PCR-C) is the premium one with obvious health benefits. In order to explore the microbiota in PCR-C and their relationship with the chemical components during aging, culture-independent methods were applied to investigate PCR-C microbiota for the first time. Here in different PCR-C samples, 16S rRNA gene amplicon sequencing revealed common central bacterial community, which were absent or only accounted for small proportion in fresh pericarps or jute bag controls. Bacillus and Lactococcus were the top two dominant genera in PCR-C with acidic pH (4.06-4.51) and low moisture (11.48%-19.13%). Several OTUs were found to closely relate with specific compositions in essential oils and phenolics, such as d-limonene and nobiletin, which contributed to PCR-C flavor and quality. As the first study to reveal the central bacterial communities in PCR-C, it provides new insights to improve the quality and aging process of traditional Pericarpium Citri Reticulatae, and lays foundation for functional characterization of the microbes within.

Correlation between lipid-lowering efficacy and components of Pericarpium Citri Reticulatae

Based on pharmacodynamics-component correlation analysis,the best effective part of hyperlipidemia of Pericarpium Citri Reticulatae( PCR) was screened out to confirm the possible constituents with the lipid regulating effect,in order to provide a basis for the development of new drugs. Hyperlipidemia rats induced by fat emulsion were used to screen out the best hypolipidemic parts of PCR with TC,LDL-c as the index. HPLC-ESI-MS were used to analyze common constituents of the different solvent extracts from PCR. The constituents were classified and identified based on the retention time,m/z and UV spectra. And the HPLC-DAD were used to determine the contents of flavonoids( narirutin,hesperidin,didymin,nobiletin,tangeretin,3,5,6,7,8,3',4'-heptemthoxyflavone).Correlation analysis was conducted on the constituents and efficacy with the method of SPSS ANOVA bivariate correlation. Five extracts could significantly decrease the content of TC,LDL-c of hyperlipemia rats induced by fat emulsion,and the best effective part were95% ethanol and ethyl acetate. There were 19 common peaks in five different solvent extracts from PCR,and 17 flavonoids were identified and classified,including 10 polymethoxyflavonoids and 7 other flavonoids. According to the raw material quantity,the order of content of flavonones arranged from high to low: n-butyl alcohol part> 95% ethanol part> water part> ethyl acetate part> petroleum ether part; and the order of PMFs arranged from high to low: n-butyl alcohol part≈95% ethanol part≈ethyl acetate part > petroleum ether part > water part. The decreased percentage of TC,LDL-c was positively correlated with 10 common PMFs constituents,which suggested that PMFs may be the effective components for reducing blood lipid.

A simple method to discriminate Guangchenpi and Chenpi by high-performance thin-layer chromatography and high-performance liquid chromatography based on analysis of dimethyl anthranilate

Citri Reticulatae Pericarpium (CRP), the dried pericarp of Citrus reticulata Blanco, can be divided into “Guangchenpi” (GCP, the dried pericarps derived from Citrus reticulata ‘Chachi’) and “Chenpi” (CP, the dried pericarps derived from other cultivars of Citrus reticulata Blanco). To discriminate between GCP and CP, a simple and reliable high-performance thin-layer chromatography (HPTLC) method was firstly developed to analyze the volatile compound dimethyl anthranilate, and a high-performance liquid chromatography (HPLC) method was established to simultaneously quantify dimethyl anthranilate and three predominant flavonoids (hesperidin, nobiletin and tangeretin) in CRP samples. Both the HPTLC analysis and HPLC-orthogonal partial least squares discrimination analysis (OPLS-DA) indicated that GCP can be effectively distinguished from CP based on analysis of dimethyl anthranilate. Our results indicated that dimethyl anthranilate can be used as a marker compound for discrimination of GCP and CP. This work provided a convenient approach which might be applied for quality evaluation of CRP.

The Chinese Herbal Formula Shenzhu Tiaopi Granule Results in Metabolic Improvement in Type 2 Diabetic Rats by Modulating the Gut Microbiota.

Objective The aim of this study is to investigate the implication of the Chinese herbal formula (CHF) Shenzhu tiaopi Granule (STG) in type 2 diabetes mellitus (T2DM) and discuss the mechanisms by which STG regulates the gut microbiota. Method Goto-Kakizaki (GK) rats and age-matched Wistar (W) rats were acclimatized for 1 week. The GK rats were randomly divided into 3 groups and orally gavaged with saline (model group, M), acarbose (acarbose group, A), and STG (granule of CHF group, G; the component of this formula includes Codonopsis pilosula, Rhizoma Atractylodis, Pinellia, Poria cocos, Pericarpium Citri Reticulatae, Coptis chinensis Franch, and Pueraria). The W rats were orally gavaged with saline (control group, C). The observation time was 8 weeks. The weight, fasting blood glucose (FBG) level, and blood lipid levels were tested. The 16S rRNA genes in the V3-V4 region were sequenced, and the structure of the gut microbiota was analysed. Results Compared to C, M displayed significant differences in blood glucose, gut microbiota, etc. (P<0.05; P<0.01). Compared to M, A and G showed a similar reduction in the FBG gain and a shift in the structure of the gut microbiota (P<0.05; P<0.01). Compared with A, G exhibited a significant decrease in weight, FBG level, and total cholesterol (P<0.05). The gut microbiota, Bacteroidetes, the Firmicutes/Bacteroidetes ratio, Allobaculum, and Desulfovibrionaceae were significantly decreased in response to the STG treatment, while Lactobacillus was significantly enriched (P<0.05; P<0.01). The community composition also differed at the phylum and genus levels based on the linear discriminant analysis effect size and heat map. Conclusion Our findings suggest that the composition of the gut microbiota was significantly changed in the diabetic GK rats compared with that in the normal W rats. STG treatment can improve glucose and lipid levels and modulate the gut microbiota in T2DM rats.

Supercritical fluid CO2 extraction of three polymethoxyflavones from Citri reticulatae pericarpium and subsequent preparative separation by continuous high-speed counter-current chromatography

Polymethoxyflavones (PMFs) are widely found in Citri Reticulatae Pericarpium (CRP) and have been investigated with a broad spectrum of biological activities as well as health promoting properties. However, separation of the PMFs from a complex sample, especially preparative separation of these PMFs with high purity, remains challenging. In the present study, an efficient method based on supercritical fluid extraction (SFE) and continuous high-speed counter-current chromatography (HSCCC) has been developed for extracting and preparative purification PMFs from CRP. Various experimental conditions were investigated to optimize the SFE and HSCCC processes. Under these optimized conditions, crude extract of CRP (extract I) was obtained with a maximum contents of nobiletin, 3,5,6,7,8,3′,4′-heptamethoxyflavone and tangeretin. Further extraction of crude extract I was carried out to obtain crude extract II, which was further isolated and purified by HSCCC. It was worth mentioned that continuous injection HSCCC process were realized without lost of separation efficiency, which allowed for multiple purification cycles and therefore saved a lot of labor and time. Furthermore, high-performance liquid chromatography (HPLC) was employed to analyze the fractions separated by HSCCC, which revealed that the purities of the three PMFs were all above 98%. The structures of the three PMFs were identified by LC–MS and 1H NMR spectroscopy.