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ObjectiveTo establish an ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry(UHPLC-QqQ-MS) for determination of the active ingredients in Erdongtang, and to predict the targets and pathways of anti-insulin resistance action of this formula. MethodThe analysis 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)-acetonitrile(B) for gradient elution(0-3 min, 90%-87%A; 3-6 min, 87%-86%A; 6-9 min, 86%-83%A; 9-11 min, 83%-75%A; 11-18 min, 75%-70%A; 18-19 min, 70%-52%A; 19-22 min, 52%A; 22-25 min, 52%-5%A; 25-27 min, 5%-90%A; 27-30 min, 90%A). The contents of active ingredients in Erdongtang was detected by electrospray ionization(ESI) and multiple reaction monitoring(MRM) mode under positive and negative ion modes. On this basis, network pharmacology was applied to predict the targets and pathways of Erdongtang exerting anti-insulin resistance effect. ResultThe 20 active ingredients in Erdongtang showed good linear relationships within a certain mass concentration range, and the precision, stability, repeatability and recovery rate were good. The results of determination showed that the ingredients with high content in 15 batches of samples were baicalein(1 259.39-1 635.78 mg·L-1), baicalin(1 078.37-1 411.52 mg·L-1), the ingredients with medium content were mangiferin(148.59-217.04 mg·L-1), timosaponin BⅡ(245.10-604.89 mg·L-1), quercetin-3-O-glucuronide(89.30-423.26 mg·L-1), rutin(46.91-1 553.61 mg·L-1), glycyrrhizic acid(55.97-391.47 mg·L-1), neomangiferin(37.45-127.03 mg·L-1), nuciferine(0.89-63.48 mg·L-1), hyperoside(6.96-136.78 mg·L-1), liquiritin(30.89-122.78 mg·L-1), liquiritigenin(26.64-110.67 mg·L-1), protodioscin(58.57-284.26 mg·L-1), the ingredients with low content were wogonin(7.16-20.74 mg·L-1), pseudoprotodioscin(5.49-22.96 mg·L-1), ginsenoside Rb1(7.31-23.87 mg·L-1), ginsenoside Rg1(10.78-28.33 mg·L-1), ginsenoside Re(7.78-24.76 mg·L-1), ophiopogonin D(2.08-4.29 mg·L-1), methylophiopogonanone A(0.74-1.67 mg·L-1). The results of network pharmacology indicated that the mechanism of anti-insulin resistance exerted by Erdongtang might be related to the phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) signaling pathway. ConclusionThe established UHPLC-QqQ-MS has the advantages of simple sample processing, strong exclusivity and high sensitivity, and can simultaneously determine the contents of the main ingredients from seven herbs in Erdongtang, which can lay the foundation for the development of Erdongtang compound preparations. The results of the network pharmacology can provide a reference for the mechanism study of Erdongtang in the treatment of type 2 diabetes mellitus.
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@#Abstract: It is well-known that crystal form of a drug is a key factor impacting the physicochemical properties of the drug, which in turn affects its in vivo efficacy, safety and stability. The study on crystal forms of solid-state drugs is crucial for drug quality control, selection of production process and evaluation of clinical efficacy. The combination of chemometric and analytical techniques exhibited its great ability to analyze a large amount of multidimensional data, providing the possibility for quantification of trace amount of crystals (< 1%). Meanwhile, using the process analytical technology (PAT) to monitor the crystal content real-time during prescription preparation process can further realize the control on formulation quality and serve as a core technology to support the patent protection of crystalline forms. In this review, the combined application of crystal analytical techniques and chemometric methods for the quantitative analysis of trace crystals were summarized, aiming to provide guidance for the manufacturing of pharmaceutical preparations and their quality control.
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OBJECTIVE To establish a quantitative analysis model for the contents of tussilagone, moisture, ethanol-soluble extract and total ash in Farfarae Flos based on near-infrared spectroscopy (NIRS), providing a new idea for the rapid quality evaluation of Farfarae Flos and its preparations. METHODS Referring to the 2020 edition of the Chinese Pharmacopoeia, the contents of the main quality control indexes tussilagone, moisture, ethanol-soluble extract and total ash in 130 batches of Farfarae Flos from 19 producing areas were determined by HPLC, drying method, hot dip method and ash assay, respectively. The NIRS data information of the medicinal herbs of Farfarae Flos was collected, and then NIRS combined with the partial least squares method was used to establish the individual quantitative analysis models of the above quality control indexes in the samples, and the predictive model of the NIRS content was obtained after sample validation with validation set. RESULTS The range for the contents of tussilagone, moisture, ethanol-soluble extract and total ash in 130 batches of Farfarae Flos were 0.051 4%-0.103 5%, 7.75%-10.93%, 20.17%-31.12%, and 7.68%-12.10%, respectively. The internal cross-validation coefficients of determination (R2) of the established models for the quantitative analysis of tussilagone, moisture, ethanol-soluble extract and total ash in Farfarae Flos were 0.985 8, 0.968 4, 0.973 4, 0.988 0, respectively; the root mean square errors of calibration (RMSEC) were 0.001 54, 0.187, 0.478, 0.127, respectively; the root mean square errors of prediction (RMSEP) were 0.001 81, 0.212, 0.543, 0.149, respectively; RMSEP/RMSEC were 1.175 3, 1.133 7, 1.136 0 and 1.173 2, respectively, which were all within a reasonable range (1<RMSEP/RMSEC≤1.2). The mean absolute errors between the true and model-predicted values of the above four quality control indexes in the validation set of samples were -0.000 36, 0.061 43, 0.144 00, and 0.010 43, respectively,and the mean predicted recoveries were 99.65%, 100.72%,100.66%, and 100.15%, respectively. CONCLUSIONS The established NIRS quantitative analysis model has high stability and reliable results, which can be used for the rapid batch prediction of the content of relevant quality control indexes in Farfarae Flos.
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Objective:To establish the HPLC fingerprint of Bolbostemmatis Rhizoma standard decoction; To determine the three effective components with similar structure by quantitative analysis of multi-components by single marker (QAMS); To evaluate the quality of Bolbostemmatis Rhizoma standard decoction.Methods:HPLC was adopted to establish the fingerprints of 15 batches of Bolbostemmatis Rhizoma standard decoction. The Chromatographic column was Waters XBridge Phenyl (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution. Cluster analysis (HCA) and principal component analysis (PCA) were conducted based on the relative peak area of common peaks. The same method as the fingerprint was used to establish QAMS of tubeimoside A, B, C on Bolbostemmatis Rhizoma standard decoction.Results:There were 14 common peaks in the fingerprint of Bolbostemmatis Rhizoma standard decoction. It was confirmed that the peak 3 was L-tryptophan, the peak 11 was tubeimoside B, the peak 12 was tubeimoside C, and the peak 13 was tubeimoside A. 15 batches of Bolbostemmatis Rhizoma standard decoction from different origins were divided into 3 categories by HCA and PCA. There was no significant difference between QAMS and the external standard method (ESM) through the system suitability inspection. Conclusion:This method is accurate, reliable and has good specificity, which can effectively evaluate the quality of Bolbostemmatis Rhizoma standard decoction.
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BACKGROUND:Total knee arthroplasty is one of the effective methods to treat end-stage knee osteoarthritis.However,some patients still experience chronic post-surgical pain.It is significant to find out the influencing factors of chronic post-surgical pain.Demographic factors,social psychological factors and perioperative pain were the focus of previous studies,but muscle factors closely related to the occurrence and development of knee osteoarthritis were rarely reported. OBJECTIVE:To evaluate the value of preoperative quantitative ultrasound analysis of quadriceps femoris in predicting chronic post-surgical pain after total knee arthroplasty. METHODS:A total of 250 patients with knee osteoarthritis who underwent the first unilateral total knee arthroplasty under elective general anesthesia from January to August 2022 in the Affiliated Hospital of Xuzhou Medical University were selected.All patients were treated with the same anesthesia and operative methods.Before the surgery,clinical data were recorded,and the thickness and echo intensity of quadriceps femoris on the operated side were measured by ultrasound imaging,which could quantify the degree of quadriceps femoris atrophy.Multivariate logistic regression was used to analyze the independent factors affecting the occurrence of chronic post-surgical pain,and receiver operating characteristic curves were used to evaluate its predictive value. RESULTS AND CONCLUSION:(1)250 subjects were involved in the result analysis,and 91 of them had chronic post-surgical pain,with an incidence of 36.4%.(2)There were significant differences between the chronic pain and non-chronic pain groups in preoperative pain score during movement,preoperative Western Ontario and McMaster University Osteoarthritis Index,preoperative anxiety and depression scale score,preoperative muscle thickness and echo intensity of quadriceps femoris,and postoperative acute pain score(P<0.05).(3)Multivariate logistic regression analysis showed that preoperative thickness of quadriceps femoris was an independent protective factor for chronic post-surgical pain and preoperative pain score during movement was an independent risk factor for chronic post-surgical pain.(4)Receiver operating characteristic curves showed that the area under the curve of the preoperative thickness of quadriceps femoris was 0.625(95%CI:0.555-0.695),and the critical value was 2.78 cm,sensitivity was 0.802,specificity was 0.415.(5)It is concluded that the preoperative thickness of quadriceps femoris is an independent protective factor for chronic post-surgical pain,but its predictive efficacy is low,and its clinical application needs to be further verified or modified.
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Objective Based on the Hood Policy Tool perspective on China's DIP policy text mining,word frequen-cy analysis and clustering analysis of the policy content,to explore the actual policy in each pilot region,to further analyze the reasons,to provide a scientific and reference for the subsequent policy adjustment and implementation.Methods Policies were included in this study from platforms such as local healthcare protection bureaus,local health-care commissions,local people's government portals,and NVivo,from which policy documents with a high degree of relevance to DIP were selected.NVivo 20.0 software was used for text coding,word frequency analysis,and analysis based on Hood's policy tool dimensions and regional distribution dimensions to further explore DIP policy specifics.Results A total of 27 policies were included,covering all pilot provinces,with a total of 1,707 policy codes.Authoritative,fiscal,informational,and organizational policy instruments accounted for 54.9%,20.3%,16.9%,and 7.9%,respectively.The percentage of coded reference points in the East,Central,West,and Northeast regions were 23.0%,20.2%,44.5%,and 8.7%,respectively.Conclusion The use of DIP policy tools in the pilot regions focused on authoritative tools and less on organization tools.Among the authoritative policy tools,the sub-tools of mechanism building were mostly used,while the sub-tools of standards and norms were less used.The focus of the policy tools varies among the pilot regions,and the use of policy tools should be tailored to the local context.
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AIM To establish a quantitative analysis of multi-components by single-marker(QAMS)method for the simultaneous content determination of gastrodin,parishin E,syringin,parishin B,parishin C,ferulic acid,parishin A,buddleoside,harpagoside and cinnamic acid in Tianma Toufengling Capsules.METHODS The analysis was performed on a 30℃thermostatic GL Science InertsilTM ODS-3 column(150 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,280 nm.Syringin was used as an internal standard to calculate the relative correction factors of the other nine constituents,after which the content determination was made.RESULTS Ten constituents showed good linear relationships within their own ranges(r≥0.999 7),whose average recoveries were 98.53%-102.22%with the RSDs of 1.26%-2.68%.The result obtained by QAMS approximated those obtained by external standard method.CONCLUSION This accurate and specific method can be used for the quality control of Tianma Toufengling Capsules.
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Objective:To apply digital analysis to quantify hyperechogenicity of substantia nigra, and explore its clinical value for diagnosis of Parkinson′s disease (PD).Methods:The cross-sectional study included 652 PD patients (PD group) and 99 healthy controls (healthy control group) from November 2017 to October 2020 in Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology. All subjects underwent transcranial sonography. The diagnostic accuracy of substantia nigra hyperechogenicity using digital analysis was compared with that in a manual measurement in PD. Furthermore, the receiver operating characteristic (ROC) curve analysis was applied to explore its diagnosis value in PD.Results:There were 482 subjects including 400 in the PD group and 82 in the healthy control group, whose quantified results of substantia nigra hyperechogenicity could be used for analysis. The ROC analysis showed that the area under the curve of the quantified larger substantia nigra hyperechoic region detection for diagnosing PD was 0.858 (95% CI 0.805-0.910), the sensitivity was 87.8%, and the specificity was 73.2%, consistent with that of doctors (area under the curve: 0.884). Further more, among these PD patients, there was no correlation between larger substantia nigra hyperechogenicity and age, age of onset, course of disease, non-motor symptoms, and motor symptoms (all P>0.05). Conclusions:Digital analysis was used to quantify the changes in substantia nigra hyperechogenicity in this seudy. The results showed that diagnostic accuracy for PD based on digital analysis was consistent with that of experienced clinicians.
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ObjectiveTo investigate the influence of concentration ratio(CR) between the internal reference and target components on the quantitative accuracy of quantitative analysis of multi-components by single marker(QAMS) by taking ginsenosides as an example. MethodUltra performance liquid chromatography(UPLC) was employed, the contents of nine components in Ginseng Radix et Rhizoma(ginsenosides Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rb3, Rd) were determined by external standard method(ES). Using ginsenoside Rg1 as the internal reference, the contents of the remaining 8 ginsenosides were determined by QAMS, and the quantitative results were compared with those of ES to evaluate the quantitative accuracy of the established QAMS. According to the contents of these 9 ginsenosides, the simulated sample solutions with different CRs of ginsenoside Rg1 to ginsenosides Rf, Rb2, Rd were prepared with the reference substance(CR=100∶1, 10∶1, 5∶1, 2∶1, 1∶1, 0.5∶1, 0.25∶1), in order to validate the effect of the CRs between the internal reference and other components on the quantitative accuracy of the QAMS. ResultThe results of ginsenosides Re, Rf, Rb1, Rc, Rb2 calculated by the two methods were the same with the relative standard deviation(RSD)<3%, however, the results of ginsenosides Rh1, Rb3 and Rd calculated by the two methods were different with RSDs of 7.06%-9.61%. According to the result of the simulated sample solution, the difference between the calculated results of ES and QAMS was large when the CR between the internal reference(ginsenoside Rg1) and other components was 5 or 10 or even higher. ConclusionThe quantitative error of QAMS will increase when the CR between the quantitative component and the internal reference is too large, so it is suggested that when establishing the QAMS, the components with close concentration to the internal reference should be selected for quantification.
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OBJECTIVE To extract and isolate the four chemical components of Yao medicine Ventilago leiocarpa, and to conduct identification and content determination for them. METHODS The chemical components of V. leiocarpa were separated and purified by solvent extraction, extraction, silica gel column chromatography and preparative liquid chromatography; then the chemical structures of four isolated compounds were identified based on their spectral data. The contents of four components were determined by high performance liquid chromatography(HPLC)-quantitative analysis of multi-components by single-marker (QAMS) method, with the following chromatographic conditions: chromatographic column was Echway GowonTM C18 (250 mm× 4.6 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution; the detection wavelength was 269 nm, and the column temperature was 25 ℃ . Using emodin as internal reference, the relative correction factors (fi/s) between emodin and the other 3 components were established and used to calculate the content. At the same time, the content of each component was calculated with the external standard method (ESM), and the differences between these two methods were compared. RESULTS Four compounds were isolated from V. leiocarpa, and they were identified as emodin, frangulin A, pleuropyrone A, emodin-8-O-β-D-glucoside. The result of HPLC-QAMS showed that the fi/s of pleuropyrone A, emodin-8-O-β-D- glucoside and frangulin A were 1.147 2, 0.874 7 and 0.644 4, respectively. The content of these four components was measured as a good linearity (r≥0.999 6); relative standard deviation (RSD) of precision, stability and reproducibility tests were all lower than 2.00%, and average recoveries were E-mail:dearhuangjianyou@126.com 99.41%-100.46%(RSD≤2.05%). There was no significant difference between QAMS method and ESM (RSD<3.00%). CONCLUSIONS Emodin, frangulin A, pleuropyrone A and emodin- 8-O-β-D-glucoside are isolated from V. leiocarpa; among them, the last three components are all isolated from for the first time. The established HPLC-QAMS method is accurate and reliable for the determination of 4 components in V. leiocarpa, and can used for quality control of V. leiocarpa.
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OBJECTIVE To sort out reform policy for basic medical insurance designated retail pharmacy (referred to as designated retail pharmacy) in China, and to provide reference for the improvement of the policy under the new situation of mutual- aid mechanism for covering outpatient bills. METHODS The policy texts on designated retail pharmacies issued by ministries and commissions of the State Council and departments directly under the State Council were collected from December 1998 to June 2023. The content analysis and social network analysis were adopted to construct a three-dimensional analytical framework based on the policy subject, the policy tool, and the policy process, in order to quantitatively analyze the policies on reforming designated retail pharmacies. RESULTS & CONCLUSIONS The reform policy of designated retail pharmacies can be roughly divided into three stages: germination, exploration and development, and in-depth promotion. The use of policy tools is dominated by environment-oriented tools, and the cooperation network of policy subjects presents a “head-body-tail” chain structure. With the advancement of policy reforms, the number of policy subjects showed a trend of decline followed by growth, the number of policy documents showed an upward trend, emphasizing the use of tools such as the construction of the pharmacist system, the flow of prescriptions, the payment of medical insurance, and the management of “dual-channels” and “outpatient co-ordination”. It is suggested that in terms of policy formulation, all policy subjects should adhere to top-level design, grasp the characteristics of the stage of policy development, and adjust the use of policy tools according to local and timely conditions; we should also strengthen cooperation and communication, improve policy formulation efficiency, achieve policy coordination, and continuously improve policies for designated retail pharmacies.
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ObjectiveTo establish a rapid and stable liquid chromatography-mass spectrometry(LC-MS) for simultaneous analysis of 17 chemical components in Gnaphalium affine aboveground parts with flowers, so as to provide experimental basis for improving the quality standard of this herb. MethodUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used for the quantitative analysis of 17 constituents in 15 batches of G. affine from different origins, the separation was performed on an ACQUITY UPLC® BEH C18 column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of methanol(A)-0.1% formic acid aqueous solution(B) for gradient elution(0-1.0 min, 8%A; 1.0-4.0 min, 8%-26%A; 4.0-9.0 min, 26%A; 9.0-14.0 min, 26%-34%A; 14.0-14.5 min, 34%-45%A; 14.5-15.0 min, 45%-60%A; 15.0-18.0 min, 60%-90%A; 18.0-19.0 min, 90%A; 19.0-19.01 min, 90%-8%A; 19.01-20.0 min, 8%A), the flow rate was 0.3 mL·min-1, the column temperature was 40 ℃ and the injection volume was 2 μL. And the electrospray ionization was used with full scanning in both positive and negative ion modes, and the scanning range was m/z 100-1 000. ResultThe established method has been verified by the methodology and could be used for the simultaneous quantification of 17 components in G. affine. The content ranges of the 17 components(quinic acid, gallic acid, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 1,3-O-dicaffeoylquinic acid, isochlorogenic acid A, isoquercitrin, 1,5-O-dicaffeoylquinic acid, apigenin-7-O-glucoside, astragalin, isochlorogenic acid C, luteolin, apigenin and hispidulin) in 15 batches of G. affine samples was 39.60-179.12, 0.17-0.84, 2.41-8.38, 4.33-31.50, 13.63-180.38, 2.43-14.75, 1.16-19.68, 0.49-5.63, 55.77-445.16, 0.23-10.26, 62.04-530.10, 1.11-18.01, 11.36-90.61, 12.22-65.98, 7.22-69.84, 3.37-45.65, 0.30-2.59 μg·g-1, respectively. The content of organic acids was higher than that of flavonoids in G. affine, and the contents of 1,5-O-dicaffeoylquinic acid, isochlorogenic acid A, quinic acid and chlorogenic acid were higher. Meanwhile, the content of flavonoids in the samples from Guizhou was higher than that from Jiangsu, while the content of organic acids in the samples from Jiangsu was higher than that from Guizhou. ConclusionThe established method can be used for the rapid and accurate determination of 17 components in G. affine, which clarifies the content range of the main components in this herb, and can provide a reference for the selection of quality control markers of G. affine.
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ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang(LJZT), and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), and the resulting compounds were identified by using databases, such as MassBank, PubChem, ChemSpider, Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform(TCMSP), and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT, including liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature, database and MS information, a total of 79 compounds were identified from LJZT, including 31 flavonoids, 15 terpenoids, 14 nitrogen-containing compounds, 6 phenylpropanoids, 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges, and the precision, stability, reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5, 2.602 1, 0.082 6, 0.128 1, 1.778 6, 0.015 7, 0.006 7, 0.030 4, 0.003 2 mg·g-1, respectively. ConclusionThe established method can quickly, sensitively and accurately analyze the chemical constituents in LJZT, clarify that the material basis of LJZT is mainly flavonoids, terpenoids and nitrogen-containing compounds, and simultaneously determine the contents of the 9 components, which can lay a foundation for the research on quality control, mechanism and clinical application of LJZT.
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Objective To establish HPLC fingerprints of soaking Euodiae fructus with water decoction of Radix glycyrrhizae by Zhangbang method(hereinafter referred to as"soaking Euodiae fructus"),and to determine the content of evodiamine,rutaecarpine and evocarpine,so as to provide the basis for the quality control and standard improvement of soaking Euodiae fructus.Methods The fingerprint of soaking Euodiae fructus was established based on wavelength switching technology,and the similarity evaluation was conducted.Taking the rutaecarpine as the internal reference,the relative correction factors of evodiamine and evocarpine were calculated by slope correction method,and the differences in measurement results between quantitative analysis of multi-components by single-marker(QAMS)method and external standard method were compared.Results The established fingerprint of soaking Euodiae fructus had a total of 20 common peaks,the similarity between it and the control fingerprint spectrum was 0.970 to 0.998,and 7 common peaks of them were identified.The contents of evodiamine and evocarpine determined by QAMS method were 1.754-7.542 mg/g and 1.281-2.455 mg/g in 10 batches of samples,and the results obtained by QAMS method and external standard method were similar.Conclusion The established HPLC method is simple,reliable,with good separation and repeatability,and can be used for quality evaluation of processed Euodiae fructus.
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ObjectiveTo analyze the correlation between 11 small molecule active components and 1 protein component of characteristic processed products with porcine cardiac blood and other products of Salviae Miltiorrhizae Radix et Rhizoma(SMRR) from Menghe medical school and anti-cerebral ischemic oxidative damage, and to identify its key component markers of characteristic processed products with porcine cardiac blood for anti-cerebral ischemic oxidative damage. MethodHigh performance liquid chromatography(HPLC) was established to simultaneously determine the contents of 11 active ingredients in SMRR and its processed products[processed with porcine cardiac blood, porcine blood, wine and transferrin(Tf) in porcine cardiac blood], and the content of Tf in different processed products of SMRR was detected by enzyme-linked immunosorbent assay(ELISA). Furthermore, A zebrafish ischemic stroke model was constructed to evaluate the effects of different processed products of SMRR on the behavioral trajectory of cerebral ischemic zebrafish, the neuronal damage of transgenic zebrafish Tg(elavl3:eGFP) brain, as well as the levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in the brain tissues. The hippocampal neurons oxygen-glucose deprivation/reoxygenation(OGD/R)-induced ischemia-hypoxia model was constructed to evaluate the effects of different processed products of SMRR on oxidative damage of neuronal cells by taking lactate dehydrogenase(LDH), reactive oxygen species(ROS), MDA and SOD as indexes. Finally, principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and Spearman correlation analysis were used to analyze the 11 small molecule active components and 1 protein component with efficacy indicators, in order to screen the key components of the characteristic processed products with porcine cardiac blood for cerebral ischemic oxidative damage. ResultCompared with the raw products, the contents of water-soluble and fat-soluble components in processed products of SMRR increased to different degrees, while the content of salvianolic acid A decreased. Compared with the wine-processed products, the contents of salvianolic acid B, danshensu, rosmarinic acid and other components in the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products were increased, while the content of salvianolic acid A was decreased. ELISA results showed that there was no significant difference in Tf content between the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products. Pharmacological results showed that different processed products of SMRR could improve the behavioral deficits, brain neuronal injury and oxidative stress after ischemic stroke in zebrafish, and the effect of the porcine cardiac blood-processed products was most pronounced. PCA results showed that salvianolic acid B, salvianolic acid A, rosmarinic acid, lithospermic acid, danshensu, tanshinone ⅡA, caffeic acid, cryptotanshinone and tanshinone Ⅰ were the main contributing components of SMRR and its processed products. And the results of correlation analysis showed that the contents of cryptotanshinone, rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and tanshinone Ⅰ were negatively correlated with MDA level in zebrafish brain tissue, while the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B and Tf were positively correlated with SOD level, and the contents of rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA, tanshinone Ⅰ, danshensu, Tf were positively correlated with neuronal fluorescence intensity in the zebrafish brain. And the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and Tf were negatively correlated with LDH, ROS and MDA levels and positively correlated with SOD level. ConclusionThere are differences in the anti-ischemic oxidative damage effects of SMRR and its different processed products, among which the porcine cardiac blood-processed products has the strongest effect on improving oxidative damage, which may be related to the content changes of salvianolic acid B, danshensu, rosmarinic acid and other components. This study can provide a basis for clarifying the quality markers of SMRR processed with porcine cardiac blood for cerebral ischemia and elucidating its processing mechanism.
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Natural colorants derived fom plant materials have gained increasing popularity due to their non toxic nature. pigment extraction from the florets is normally done by Soxhlet extraction, maceration, and hydro distillation are conventional methods that have been widely used in industry and laboratory .phytochemical analysis of safflower florets revealed the plant presence of high amount of Carthamin and carthamidin.
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The article is devoted to the study of lipids and fatty acid composition of the above-ground part of the Nepeta olgae Regel (L.) plant of the Laminaceae family. It was found that the content of neutral lipids (NL) is 5.54%, PL - 6.12%, and total lipids (NL, PL) - 11.66%. Of the neutral lipids, the unsaponifiable substances (HB) had a bright yellow color, which is explained by a small amount of carotenoids (88.87 mg%). Glycolipids dominate in PL. Among the unsaponifiable substances were found biologically active components such as hydrocarbons, carotenoids, aliphatic alcohols, sterols and triterpenols. Phytosterols were the main component of unsaponifiable NS. Quantitative and qualitative analysis of fatty acids from the plant Nepeta olgae Regel (L.) was carried out by gas chromatography (GC). 28 acids were identified, of which 11 compounds are saturated, and 7 compounds are unsaturated fatty acids. Of the fatty acids, the main ones are linolenic 18:3 (35.48), palmitic 16:0 (33.38%), as well as ?-3 polyunsaturated fatty acids, including eicosanoic 20:1, cis-11,14-eicosadienoic 20:2, 8,11,14-eicosatriene 20:3 + arachidonic 20:4. Extracts of Nepeta olgae Regel (L.) were distinguished by a high content of polyunsaturated acids, which determines their potential biological activity.
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This article for the first time presents the results of the study of qualitative and quantitative elemental and amino acid composition of the aboveground part of the plant Nepeta olgae Regel (L.) taken in the territory of Chust and Kosonsai districts (from the slopes of Gova and Kosonsai mountains) of Namangan region during the period before and during flowering (May-June, 2021-2022). The use of instrumental analysis of high-throughput energy dispersive X-ray fluorescence spectrometry, allowed to establish 20 mineral elements in the plant Nepeta olgae Regel (L.), among which to vital 9 elements and 3 to conditionally necessary. The amino acid composition of the plant Nepeta olgae Regel (L.) was studied by high performance liquid chromatography (HPLC) and 17 compounds were identified. Of these, 8 were substitutable and 9 essential amino acids.
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Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.
Assuntos
Humanos , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Medicamentos de Ervas Chinesas/química , Controle de Qualidade , Artrite GotosaRESUMO
@#Facial symmetry evaluation has always been a hot topic of concern for doctors who engage in the study of facial beauty disciplines such as orthodontics, dentistry, and plastic surgery. Although scholars at home and abroad have carried out much research on the evaluation of facial symmetry with a variety of emerging technologies and methods, there is still a lack of unified standards for the evaluation of facial asymmetry due to the complexity of the content and methods and individual subjectivity. Facial asymmetry involves changes in the length, width and height of the face. It is a complex dental and maxillofacial malformation whose early identification and accurate evaluation are particularly important. Clinically, in addition to the necessary dental and maxillofacial examinations, it is also necessary to evaluate facial asymmetry with the help of corresponding auxiliary methods. This paper gives a summary of the commonly used three-dimensional evaluation methods. The evaluation methods of facial asymmetry can be divided into 5 categories: qualitative analysis, quantitative analysis, dynamic analysis, mathematical analysis, and artificial intelligence analysis. After the analysis and summarization of the characteristics, advantages and limitations of each method in clinical applications, it is found that although these methods vary in accuracy, evaluation scope, diagnosis nature and calculation method, etc., the three-dimensional evaluation methods are more objective, more accurate and more convenient and will become the mainstream evaluation method for facial asymmetry with further development of three-dimensional measurement technologies.