Analysis of Chemical Components and Tissue Distribution of Lujiao Formula Based on UPLC-Q-Orbitrap-MS / 中国实验方剂学杂志

ObjectiveTo qualitatively analyze the chemical constituents and their tissue distribution in Lujiao formula based on ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Orbitrap-MS）. MethodThe separation was performed on an ACQUITY UPLC® BEH C18 column（2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution（A）-methanol（B） in a gradient elution（0-2 min， 4%B； 2-6 min 4%-12%B； 6-38 min， 12%-70%B； 38-38.5 min， 70%B； 38.5-39 min， 70%-95%B； 39-43 min， 95%B； 43-43.1 min， 95%-4%B； 43.1-45 min， 4%B）， the flow rate was 0.3 mL·min-1 with the column temperature of 40 ℃ and the injection volume of 5 µL. The data were acquired by an electrospray ionization（ESI） in the full scanning mode of positive and negative ions， the scanning rang was set at m/z 100-1 500， the collision energies were 10， 20， 40 eV. Retention time， precise relative molecular mass and secondary mass spectrometry fragment ions were used to identify the compounds in Lujiao formula and analyze their tissue distribution， combing with self-established database and comparing with standard substances and published literature data. ResultA total of 260 compounds， including 156 flavonoids， 43 terpenoids， 18 coumarins， 13 organic acids， 7 phenylethanoids， 7 alkaloids and 16 others， were identified or hypothesized from Lujiao formula， 68 of which were identified by comparison with standard substances. And the results of tissue distribution showed that 100， 143， 129 and 126 prototype components were detected in blood， heart， liver and kidney， respectively. ConclusionThe chemical composition of Lujiao formula and their tissue distribution were systematic analyzed， which can provide reference for the quality control， clinical application， pharmacokinetics and pharmacodynamic material basis of Lujiao formula and its medicinal materials.

Analysis of Suitable Processing Time of Rehmanniae Radix Praeparata Processed with Amomi Fructus and Citri Reticulatae Pericarpium Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

ObjectiveTo investigate the relative content changes of differential metabolites and reducing sugars during the processing process of Rehmanniae Radix Praeparata （RRP） processed with Amomi Fructus （AF） and Citri Reticulatae Pericarpium （CRP）， and to lay the foundation for revealing the processing principle of this characteristic variety. MethodThe samples of the 0-54 h processing process of RRP processed with AF and CRP were taken as the research object， and their secondary metabolites were detected by ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS）. The 0.1% formic acid aqueous solution （A）-acetonitrile （B） was used as the mobile phase for gradient elution （0-1 min， 1%-3%B； 1-10 min， 3%-9%B； 10-15 min， 9%-12%B； 15-22 min， 12%-18%B； 22-31 min， 18%-24%B； 31-35 min， 24%-100%B； 35-36 min， 100%-5%B； 36-40 min， 5%-1%B； 40-45 min， 1%B）， column temperature was 40 ℃， injection volume was 3 μL， flow rate was 0.3 mL·min-1. Electrospray ionization （ESI） was used to scan and collect MS data in the negative ion mode， the scanning range was m/z 50-1 250. Data analysis was carried out using PeakView 1.2 software， and the chemical composition of RRP processed with AF and CRP was identified by combining the literature information and chemical composition databases. The MS data were normalized by MarkerView 1.2， and then the multivariate statistical analysis was applied to screen the differential metabolites， and the changes of the relative contents of the differential metabolites with different processing times was analyzed， finally， correlation analysis was performed between the differential metabolites， the change of the reducing sugar content was combined to determine the most suitable processing time of RRP processed with AF and CRP. ResultA total of 121 compounds were identified from RRP processed with AF and CRP at different processing times， and 12 differential metabolites were screened out by multivariate statistical analysis， including catalpol， hesperidin， isoacteoside， acteoside， narirutin， echinacoside， isomartynoside， decaffeoylacteoside， 6-O-E-feruloylajugol， dihydroxy-7-O-neohesperidin， jionoside D， and rehmapicroside. With the prolongation of processing time， the relative contents of these 12 differential metabolites and reducing sugars changed slightly at 52-54 h. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents of RRP processed with AF and CRP at different processing times， and the suitable processing time of 52-54 h is determined according to the content changes of different metabolites and reducing sugars， which provides a basis for revealing the scientific connotation of the processing principle of this variety.

Quality Difference Analysis of Raw and Steamed Products of Citri Reticulatae Pericarpium Based on Electronic Eye Technique and Chemical Fingerprint / 中国实验方剂学杂志

ObjectiveTo explore the quality differences between steamed products and raw products of Citri Reticulatae Pericarpium（CRP）. MethodThe color of steamed products and raw products of CRP was determined from the perspective of appearance by electronic eye technique， and the quality differences between them was objectively characterized by the luminous value（L*）， yellow-blue value（b*）， red-green value（a*） and total chromatic value（E*ab）. Based on this， ultra-high performance liquid chromatography（UPLC） was used to establish a fingerprint evaluation method with the mobile phase of acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-5 min， 5%A； 5-30 min， 5%-20%A； 30-60 min， 20%-52%A）， detection wavelength at 270 nm， flow rate of 0.3 mL·min-1 and column temperature of 30 ℃. The quality differences between steamed products and raw products of CRP were compared from the perspective of chemical composition， and correlation analysis was used to reveal the correlation between the difference in appearance color and the difference in internal chemical composition. ResultAfter being steamed， L*， b* and E*ab of CRP showed an overall decreasing trend， indicating that the color of the steamed products darkened and deepened from yellow to blue but still tended to be yellow， while a* showed an overall increasing trend， indicating that the color of the steamed products tended to red. A total of 24 peaks were identified in the fingerprint profiles of raw products and steamed products of CRP， and 13 of the main peaks were identified. The precision， stability and repeatability studies showed that compared with the reference peak （peak 14， hesperidin）， the relative standard deviations（RSDs） of the relative peak area and relative retention time of the remaining peaks were<3.0%.The results of chemometric statistical analysis showed that there were some differences between raw products and steamed products of CRP， and 7 main differential components were identified， among which 5-hydroxymaltol（peak 1） and 5-hydroxymethylfurfural（peak 2） were the characteristic components of steamed products. The correlation analysis results showed that， in addition to the above two characteristic components， four components of peak 4， peak 10 （vicenin-2）， peak 23 （tangeretin） and peak 24 （5-demethylnobiletin） also correlated significantly with the color change （E*ab） of the samples （P<0.05， P<0.01）. ConclusionBefore and after steaming， not only the chemical composition changes， but also the color. Comparing the characteristic peaks of chemical composition difference and color difference before and after steaming of CRP， it is found that 5-hydroxymaltol， 5-hydroxymethylfurfural and peak 4 are common characteristic difference components， which can provide a reference for establishing the characteristic quality control method of steamed products， and quickly evaluating the quality difference between raw products and steamed products of CRP.

Analysis on Components Absorbed into Blood of Citri Reticulatae Pericarpium in Rats Based on Sequential Metabolism and UPLC-HRMS / 中国实验方剂学杂志

ObjectiveTo study the metabolism of chemical components from Citri Reticulatae Pericarpium（CRP）in different parts of rats by sequential metabolism and ultra performance liquid chromatography-high resolution mass spectrometry（UPLC-HRMS）. MethodSD male rats were employed as experimental subjects， and blood samples of intestinal metabolism and hepatic metabolism were prepared after administration of CRP ethanol extract by in situ intestinal perfusion， and comprehensive metabolic samples were collected after intragastric administration. UPLC-HRMS was used to analyze the samples with acetonitrile（A）-0.1% formic acid aqueous solution（B）as the mobile phase for gradient elution（0-10 min， 10%-30%A； 10-30 min， 30%-95%A； 30-31 min， 95%-10%A； 31-35 min， 10%A）at a flow rate of 0.35 mL·min-1， using a heated electrospray ionization with positive and negative ion mode scanning in the range of m/z 100-1 500. Under these conditions， the differences in the profiles of CRP ethanol extract， blank plasma and drug-containing plasma under different treatment groups were compared， and the chemical components of each sample were analyzed and identified based on the retention time， accurate relative molecular mass， primary and secondary ion fragments， and the information of reference substances. ResultA total of 44 chemical components were identified in the CRP ethanol extract， including flavone-O-glycosides， flavone-C-glycosides and polymethoxyflavonoids， etc. The results of sequential metabolism showed that 22 chemical components in CRP were detected in the intestinal metabolic sample， 18 chemical components were detected in the hepatic metabolic sample， and 9 identical chemical components（narirutin， hesperidin， meranzin， 5，7，8，3ʹ，4ʹ，5ʹ-hexamethoxy-flavone， isosinensetin， sinensetin， 3，5，6，7，8，3ʹ，4ʹ-heptamethoxyflavone， nobiletin and tangeretin）could be detected in all three metabolic samples， with a total of 22 compounds entering the blood in prototype form. ConclusionThe identified 21 components with well-defined structures entering the blood as prototypes may be potential active components of CRP， and differences in the components at different metabolic parts can provide an experimental basis for elucidating the in vivo biotransformation process of the metabolic components of CRP.

Processing of Citri Reticulatae Pericarpium： A Review / 中国实验方剂学杂志

Citri Reticulatae Pericarpium is a commonly used traditional Chinese medicine in China. By referring to herbal monographs and related research documents， and comparing all editions of Chinese Pharmacopoeia and local processing specifications， the processing history， modern processing specifications， study on chemical constituents and pharmacological effects before and after processing of Citri Reticulatae Pericarpium were summarized. It was found that the processing methods of Citri Reticulatae Pericarpium in the past dynasties were mainly cleansing， cutting and frying. The processing methods of Citri Reticulatae Pericarpium included in all editions of Chinese Pharmacopoeia were all cleansing and cutting. In modern local processing specifications， there were mainly processing methods such as frying， steaming and carbonizing. The components of Citri Reticulatae Pericarpium were mainly volatile oil and flavonoids. After processing， the total amount of volatile oil generally decreased， and the contents and compositions of volatile oil and flavonoids also changed. It may be the main reason for the difference of efficacy before and after processing. At present， the optimization of processing technology of Citri Reticulatae Pericarpium mainly focuses on steaming， processing with vinegar and baking. Pharmacological research on Citri Reticulatae Pericarpium mainly focuses on reducing blood lipid， relieving cough， relieving asthma and resolving phlegm， and inhibiting pulmonary fibrosis. The current research on the processing methods of Citri Reticulatae Pericarpium is not in-depth enough， the material basis of many processing methods has not been clarified， and the research on the efficacy before and after processing is not in-depth enough. Further research is needed to clarify the material basis and mechanism after the processing of Citri Reticulatae Pericarpium， so as to standardize the processing method and establish specific quality standards.

Contributions of flavonoids from citri reticulatae pericarpium to gastric hormones, CD3~+ and TFF3 mRNA expression in rats with spleen deficiency intervened by Liujunzi Decoction / 中国中药杂志

The present study established the spectrum-effect relationship model of flavonoids in Citri Reticulatae Pericarpium(CRP) from 15 batches of Liujunzi Decoction and statistically analyzed the correlation between chemical peaks and efficacy to identify the main effective components. HPLC fingerprints of flavonoids in CRP from 15 batches of Liujunzi Decoction were established. HPLC analysis was carried out on the Venusil XBP C_(18)(L) column(4.6 mm×250 mm, 5 μm) at 30 ℃ with acetonitrile-water(containing 0.1% formic acid) as mobile phase for gradient elution, a flow rate of 1.0 mL·min~(-1), and detection wavelength of 300 nm to obtain chemical fingerprints. Additionally, the effects of flavonoids from CRP in 15 batches of Liujunzi Decoction on the content of GAS, MTL, and VIP, TFF3 mRNA expression, and percentage of CD3~+ T-cells of model rats with spleen deficiency were determined. The spectrum-effect relationship model was established by gray correlation analysis. The results showed that the main characteristic peaks with great contribution to the regulation of gastrointestinal tract were peak 16(vicenin-2), peak 63(sinensetin), peak 64(isosinensetin), peak 65(nobiletin), peak 67(3,5,6,7,8,3',4'-heptemthoxyflavone), peak 68(tangeretin), and peak 69(5-desmethylnobiletin). Therefore, there was a linear correlation between flavonoids from CRP in Liujunzi Decoction and the efficacy, and the medicinal effect was achieved by multi-component action. This study is expected to provide a new idea for exploring the material basis of the effect, i.e., regulating qi prior to replenishing qi, of CRP in Liujunzi Decoction.

Analysis and Identification of Flavonoids in Citri Reticulatae Pericarpium with Different Aging Time by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:To analyze and identify the flavonoids of Citri Reticulatae Pericarpium with different aging time by an ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS）. Method:Compounds were separated on Agilent Extend-C₁₈ column （3.0 mm×100 mm， 1.8 μm）， mobile phase was 0.1% acetic acid aqueous solution （A）-0.1% acetic acid methanol solution （B） for gradient elution （0-25 min， 5%-95%B； 25-30 min， 95%B； 30-30.1 min， 95%-5%B； 30.1-35 min， 5%B）， the flow rate was 0.4 mL·min^-1， and the column temperature was set at 30 ℃. High resolution mass spectrometry was performed with electrospray ionization （ESI）， and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. The multistage ion fragment information combined with mzCloud network database， local high resolution mass spectrometry database of traditional Chinese medicine components （OTCML）， literature information and relevant reference materials were used for accurate qualitative analysis. Result:Totally 43 flavonoids in Citri Reticulatae Pericarpium were identified， including 24 flavones， 5 flavonols， 13 dihydroflavones and 1 chalcone. The flavonoids in samples with different aging time were basically consistent in material types， but the peak area was different. According to the comparison of relative content in the peak area， it was found that the relative contents of 30 flavonoids showed an overall increasing trend with the increase of aging time. Among them， the relative contents of 24 flavonoids （such as hesperidin， diosmin， 6-demethoxytangeretin， nobiletin and tangeretin） increased significantly. There was no significant change in the relative contents of the other 13 flavonoids （such as naringenin and neohesperidin）. Conclusion:An efficient method is established in this paper to identify flavonoids in Citri Reticulatae Pericarpium with different aging time and their relative content changes rapidly and accurately. The findings provide a methodological reference for the study on pharmacodynamic material base and quality control of Citri Reticulatae Pericarpium， and it provides experimental basis that drugs processed long time ago have better effect of Citri Reticulatae Pericarpium.

Improvement effect and mechanism of ethanol extract from Citri Reticulatae Pericarpium on triglyceride in hyperlipidemia model rat / 中国中药杂志

The aim of this paper was to study the improvement effect of ethanol extract from Citri Reticulatae Pericarpium(CRP) on triglyceride of hyperlipidemia model rats, and to explore the possible mechanism. SD rats were randomly divided into normal group, model group, positive control group, and high, medium and low-dose CRP ethanol extract groups, with 10 rats in each group. During the experiment, except for the normal group that was fed with distilled water and ordinary feed, rats in the other groups were given different concentrations of alcohol and fed with high-sugar and fat diets. All rats were given free diets. While being modeled, each group was administered with 0.01 mL·g~(-1) by gavage once a day for six weeks. Blood samples were collected after two weeks, four weeks and six weeks of drug treatment. After the completion of the experiment, blood, liver and adipose tissue were collected. Triglyceride(TG), alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) in serum, TG in liver tissue and TG in fecal were detected. Free fatty acid(FFA) and triglyceride-related hydrolase, such as adipose tiglyceride lipase(ATGL), lipoprotein lipase(LPL), hepatic lipase(HL), hormone-sensitive triglyceride lipase(HSL) were detected by ELISA. The mRNA expressions of peroxisome proliferators-activated receptors(PPARγ), sterol regulatory element binding protein 1 c(SREBP-1 c) and farnesoid X receptor(FXR) were determined by RT-PCR. Compared with the model group, each administration group could reduce TG levels in serum and liver to varying degrees, reduce serum ALT, AST, ALP activities, significantly reduce free fatty acid content in serum, significantly increase triglyceride metabolism-related enzymes, including fat ATGL, LPL and liver HL content, and significantly reduced the content of fat HSL. According to the study of transcriptional regulation genes relating to triglyceride metabolism, extract from CRP could significantly increase the mRNA expressions of PPARγ and FXR. In conclusion, ethanol extract from CRP could ob-viously reduce the TG level of hyperlipidemia model rats, and might reduce plasma TG content by increasing PPARγ-LPL/ATGL and FXR-HL triglyceride hydrolysis pathways.

Prediction of Q-markers of Citri Reticulatae Pericarpium volatile oil and GC-MS based quantitative analysis / 中国中药杂志

This study was designed to predict the Q-markers of Citri Reticulatae Pericarpium volatile oil and conduct quantitative analysis by GC-MS. The common components of Citri Reticulatae Pericarpium volatile oil were detected by GC-MS. The network pharmacology approaches were utilized for constructing the component-target network and protein-protein interaction(PPI) network, followed by the GO and KEGG pathway enrichment analysis to clarify the pharmacological effects of common components. Molecular docking was conducted to observe the biological activities of common components, thus identifying the Q-markers of Citri Reticulatae Pericarpium volatile oil. The obtained Q-markers were subjected to quantitative analysis by GC-MS. The GC-MS analysis of 19 batches of Citri Reticulatae Pericarpium volatile oil revealed three common components, namely, D-limonene, γ-terpinene, and myrcene. The common components were analyzed based on network pharmacology, and the results showed that Citri Reticulatae Pericarpium volatile oil mainly acted on the core targets GABRA1, GABRA6, GABRA5, GABRA3, and GABRA2 through D-limonene and γ-terpinene, with five important pathways such as nicotine addiction and GABAergic synapse involved. The core targets were mainly distributed in olfactory region, cerebral cortex, cerebellum, basal ganglia, hippocampus, and amygdala to exert the pharmacological effects. As revealed by molecular docking, D-limonene and γ-terpinene exhibited good biological activities, so they were identified as the Q-markers of Citri Reticulatae Pericarpium volatile oil. The results of quantitative analysis showed that the volume fraction of D-limonene was within the range of 0.77-1.03 μL·mL~(-1), and that of γ-terpinene within the range of 0.04-0.13 μL·mL~(-1). The prediction of D-limonene and γ-terpinene as the Q-markers of Citri Reticulatae Pericarpium volatile oil has laid an experimental foundation for the establishment of the quality evaluation standard for Citri Reticulatae Pericarpium volatile oil.

Chemome profiling of Citri Reticulatae Pericarpium using UHPLC-IT-TOF-MS / 中国中药杂志

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.

Study on herb property expression of pungent-aromatic Citri Reticulatae Pericarpium, Caryophylli Flos, and Foeniculi Fructus based on network pharmacology of olfactory receptors / 中草药

Objective: In this study, network pharmacology techniques and methods were used to screen the material basis, targets and pathways of the interaction between the pungent-aromatic herbs (Citri Reticulatae Pericarpium, Caryophylli Flos, and Foeniculi Fructus) and olfactory receptor. To explain the molecular mechanism of the expression of pungent properties of the pungent-aromatic herb. Methods: TCMSP database was used to collect the chemical components of Citri Reticulatae Pericarpium, Caryophylli Flos, and Foeniculi Fructus, in order to screen out the effective components and predict the related targets. All the proteins related to olfactory receptors were collected from UniProt database. The selected targets and ORs proteins were uploaded to the online STRING database for protein interaction analysis. The interaction network was drawn by using the software of Cytoscape software, and the key targets was further analyzed by GO functional enrichment analysis and KEGG pathway enrichment analysis. Results: Five active components of Citri Reticulatae Pericarpium, five of Caryophylli Flos and two of Foeniculi Fructus, 224 targets and 515 olfactory receptors were obtained. Protein interaction network showed 25 targets (such as GNB2, CHRM2, CHRM3, GNGT2, ADRA2A) were the key targets for ORs of three pungent-aromatic herbs. The enrichment analysis of GO and KEGG identified 31 biological processes and 78 signaling pathways. Conclusion: Olfactory receptors are mainly activated by volatile oil components of the pungent-aromatic herb. And the classical cAMP signaling pathway may not be the only one after the olfactory receptor activated, which may be involved in the expression of pungent properties of the pungent-aromatic herb together with several other signaling pathways, so as to produce "pungent flavor".

Stoichiometric Analysis in Fingerprint of Aqueous Extract of Citri Reticulatae Pericarpium and Spectrum-effect Relationship with Diabetic Cognitive Dysfunction / 中国实验方剂学杂志

<b>Objective::To establish the HPLC fingerprint of aqueous extract of Citri Reticulatae Pericarpium, and research the spectrum-effect relationship in diabetic cognitive dysfunction rats. <b>Method::The HPLC fingerprints of 21 batches of Citri Reticulatae Pericarpium were established, with acetonitrile (A)-0.1%of phosphoric acid (B) as the flow phase for gradient elution. The detection wavelength was 270 nm. Acomparative analysis was performed for the results based on similarity evaluation, SPSS 24.0 two-dimension clustering analysis and SIMCA 14.1 principal component analysis of fingerprint software. Thediabetic rat model wasduplicated by one-time intraperitoneal injection with streptozotocin (STZ) at the dose of 50 mg·kg^-1. After successful replication of the model, thediabetic rats were given aqueous extract of Citri Reticulatae Pericarpium by gavage. The blood glucose value was determined at regular intervals. The cognitive function was analyzed with Morris water maze experiment at the 12 week. Grey relational analysis (GRA) and partial least squares regression analysis were used to study the spectrum-effect relationship. <b>Result::Totally 16 common peaks of water extract of Citri Reticulatae Pericarpium were confirmed, five substances were identified, and 21 species of Citri Reticulatae Pericarpium of different varieties and origins were divided into 2 categories. Compared with the model group, the blood glucose of rats in each group treated with water extract of Citri Reticulatae Pericarpiumwere reduced, and their cognitive function was improved.According to the grey correlation analysis and the least partial square regression analysis, the peaks No. 19, 15, 4, 17 and 6 were significantly correlated with the cognitive function of diabetic rats. <b>Conclusion::There were some differences in aqueous extract of Citri Reticulatae Pericarpium between different varieties and origins. It is convenient and fast to make a comprehensive quality evaluation of aqueous extract of Citri Reticulatae Pericarpium of different varieties and producing areas by stoichiometry, and aqueous extract of Citri Reticulatae Pericarpium can reduce the blood glucose level and improve the cognitive function of diabetic cognitive dysfunction rats, its mechanisms of action are resultedfrom the joint effects of multiple effective components, and peaks No. 19, 15, 4, 17 and 6 are components closely related to the cognitive function of diabetic rats.

Analysis of Fingerprint of Standard Decoction of Citri Reticulatae Pericarpium Based on Spectroscopic Method / 中国实验方剂学杂志

Objective:To establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium and evaluate its quality. Method:According to the preparation conditions of the standard decoction,15 batches of standard decoction of Citri Reticulatae Pericarpium were prepared.HPLC was employed to determine the content of hesperidin in this standard decoction.Ultraviolet spectroscopy(UV) and infrared spectroscopy(IR) were used to establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium.The correlation coefficient method and double index sequence analysis method were used to compare and analyze the spectra of different batches of this standard decoction. Result:The content of hesperidin in 15 batches of this standard decoction were 0.82%-2.60%,and the measured value of dry extract rate was 32.02%-46.11%.Compared with ultraviolet and infrared control fingerprint,the fingerprint similarities of the standard decoction of each batch were > 0.897 and > 0.942,respectively.The double index analysis results showed that the common peak ratio was more than 62.50%,variation peak ratio was less than 46.67%. Conclusion:The quality evaluation method established in this study can be used for systematic evaluation of standard decoction of Citri Reticulatae Pericarpium,and it can provide theoretical reference for the formulation of quality standard of Citri Reticulatae Pericarpium dispensing granules and other related preparations.

Analysis of medication patterns of Chinese patent medicine for cough / 中草药

To reveal the medication patterns of Chinese patent medicine in the treatment of cough based on the analysis of the patent status of Chinese materia medica (CMM) in the field of cough control in nearly 20 years. Patents of Chinese patent medicine in the treatment of cough were systematically searched in SIPO Platform and CNKI, association analysis and network analysis of high-frequency medicines were used to reveal the medication patterns of Chinese patent medicine in the treatment of cough with software Clementine 12.0. The results showed that Chinese medicine with highest frequency in 316 formulas was Glycyrrhizae Radix et Rhizoma, the other Chinese herbs with higher frequency were Platycodonis Radix, Ephedrae Herba, Citri Reticulatae Pericarpium, Pinelliae Rhizoma, etc. The main categories were the medicines used to treat cough and asthma, as well as to reduce phlegm. The commonly used herbal pairs were Glycyrrhizae Radix et Rhizoma-Platycodonis Radix, Glycyrrhizae Radix et Rhizoma-Citri Reticulatae Pericarpium, Pinelliae Rhizoma-Glycyrrhizae Radix et Rhizoma, Ephedrae Herba-Glycyrrhizae Radix et Rhizoma, etc. It was concluded that the medication patterns of Chinese patent medicine in the treatment of cough can be revealed based on the analysis of the herbal frequency, herbal pairs, association rules, and the network of high-frequency medicines and the theory of traditional Chinese medicine, so as to provide reference evidences for clinical medication.

HPLC fingerprints research on decoction pieces of Citri Reticulatae Pericarpium based on chemometrics methods combined with orthogonal partial least square discriminate analysis / 中草药

Objective To establish an HPLC method for the fingerprints analysis of decoction pieces of tangerine peel, so as to provide reference for the quality control of it. Methods Fingerprints of 17 batches of decoction pieces of tangerine peel were established by HPLC and evaluated by cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least square discriminate analysis (OPLS-DA). Results The method of fingerprint of decoction pieces of tangerine peel was established, the similarities were greater than 0.9. There were 25 common peaks in the HPLC fingerprint, of which variable importance projection (VIP) of 14 peaks were greater than 1. Compared with the spectrogram of reference substances, peak 13 was narirutin, peak 14 was hesperidin, peak 23 was nobiletin, peak 24 was 3,5,6,7,8,3’,4’-heptamethoxy flavone, and peak 25 was hesperetin. Conclusion This simple and reliable method can be used for the identification and quality control of decoction pieces of Citri Reticulatae Pericarpium.

Study on influence of different storage environments and packaging materials on quality of Citri Reticulatae Pericarpium / 中国中药杂志

Traditional Chinese herbs are readily contaminated by mold that produced mycotoxins which are closly related to the herbs' external factors and external environments during the storage process. In this study, Citri Reticulatae Pericarpium was used as example, and the characteristics of traits, water content, active components (naringin, hesperidin, sinensetin, naringin, tangeretin) and the accumulation of aflatoxins (AFs) were selected as the evaluation indexes. Citri Reticulatae Pericarpium was stored under different environments and packaging materials for 12 months, and then the quality changes and mildew of Citri Reticulatae Pericarpium were examined. The results showed that the color of Citri Reticulatae Pericarpium was deepened after storage, but without mildew phenomenon. Besides, the sample storage in kraft paper and woven bags had varying degrees of moth phenomenon after 12 months storage, and the water content exceeded the limit of Chinese Pharmacopoeia. In addition, the contents of the five active constituents obviously decreased, especially for hesperidin, which did not meet the pharmacopoeia standard after storage. AFs were not detected in any of the tested samples. According to the results, we conclude that low temperature and humidity environment is more suitable for the storage of Citri Reticulatae Pericarpium, and that packaging materials should be further investigated. This study is of great significance for preventing the mold to contaminate the traditional Chinese medicine and ensuring the quality, effectiveness and safety of TCMs.

Research progress on Citri Reticulatae Pericarpium, Aurantii Fructus Immaturus, and Aurantii Fructus and Q-marker predictive analysis / 中草药

Citri Reticulatae Pericarpium, Aurantii Fructus Immaturus, and Aurantii Fructus belong to the traditional Chinese medicine derived from citrus and are commonly used for regulating vital (Qi in Chinese). But these three are different in the function, efficacy and therapeutic. Based on the concept of Q-marker and the combination of pharmacodynamics, pharmacological and pharmacokinetic studies, their quality markers were predicted. The prediction study provides a theoretical basis for establishing and improving the quality standard of the medicinal materials.

Optimization of processing technology of decoction pieces of Citri Reticulatae Pericarpium by Box-Behnken response surface methodology based on CRITIC-AHP weighted evaluation / 中草药

Objective To determine the weight coefficient and optimize the processing technology of decoction pieces of Citri Reticulatae Pericarpium (CRP). Methods The contents of narirutin, hesperidin, nobiletin, 3,5,6,7,8,3’,4’-heptamethoxy flavone (HF), and hesperetin were simultaneously determined by HPLC. The weight coefficient of each component was evaluated by CRITIC-AHP (analytic hierarchy process) methods. With composite score as index, Box-Behnken design-response surface methodology was adopted to investigate the effects of water addition, soaking time, and soaking temperature on the quality of the processed products and optimize the processing technology of decoction pieces of CRP. Results Optimal processing parameters were as follows: 33% of water was added to 1 000 g of herbs, soaking time was 64 min, and soaking temperature was 45 ℃. Conclusion The optimized processing technology is simple and feasible, which can provide a reference for the processing of decoction pieces of CRP. The method established to simultaneously determine the contents of five components in decoction pieces of CRP is rapid and reliable for controlling the quality of decoction pieces of CRP.

Metabolic and excretory process of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium in rats' urine and feces analyzed by RRLC-QqQ-MSn / 中国中药杂志

This present study was to investigate the metabolism and excretion of characteristic polyphenols such as flavonoids and coumarins in urine and feces of rats after intragastric administration of ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. The urine and feces of rats were collected after intragastric administration of 70% ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. Rapid resolution liquid chromatography coupled with quadrupole tandem mass spectrometry (RRLC-QqQ-MSn) was applied to compare the contents of polyphenols in ethanol extract, urine and feces. By comparing with reference substance, 30 polyphenols were identified from the ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium, including flavone glycosides, flavones, flavonone glycosides, flavonones, flavonol glycosides, polymethoxyflavones, coumarins, and limonoids and so on. The detection of various types of compounds showed differences in contents between the intestinal metabolism and excretion in the feces after systemic circulatory metabolism and renal excretion. The results showed that the polymethoxyflavones and flavonones were primarily excreted through urine, and the flavonone glycosides and limonoids were primarily excreted through feces. However, coumarins were hardly detected in feces and urine, indicating that coumarins may be metabolized in the body.

Separation and Identification of Surface Preponderant Fungi on Citri Reticulatae Pericarpium and Its Influence on Effective Substance / 世界科学技术-中医药现代化

This paper studied the influence ofsurface preponderantfungi on main effective substances of CitriReticulatae Pericarpium (CRP).Spreading plate method was used to isolatepredominant strains from the surface of CRP.Besides,the method combining microscopic and molecular identification was also adopted.HPLC and UVspectrophotometric methods were used to determine the main effective substances in CRP.From thesurface of CRP,the advantage strain fungi were Aspergillusniger and A.Flavus.After the inoculation of A.Niger and A.flavus against CRP,the effective compositionwas changed.And different A.niger strains had differenceeffectiveness oneffective chemical components,especially one strain of A.niger.Compared withthe control group,contents of total flavonoids and hesperidin were significantly increased (P＜0.01);and five types of obvious new chemical compositions were produced.It was concluded that the metabolic transformation of fungi was related tochanges ofeffective substances of CRP,which played a significant role in the aging process of CRP.The growth and metabolism of fungi consumeeffeetive substances and producechanges of composition.From the perspective of microorganisms,"the older,the better" of CRP hasa better explanation.