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1.
J Chromatogr A ; 1666: 462862, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-35124358

RESUMO

Deep profiling of chemicalome in Chinese medicinal formulas is vital for disclosing the secret underlying their effectiveness. To address this issue, an in-house database-driven untargeted identification strategy was proposed with the use of ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry. Firstly, an in-house mass spectral database for the analyzed herbs was constructed, and database querying was performed for rapid recognition of known compounds. Secondly, a chemical diagnostic characteristics algorithm was originally developed for deep mining unrecorded ions, and thus expanding coverage of components beyond the database. Additionally, we proposed evaluation criteria for the untargeted identification of compounds with different confidence levels. As a case study, the integrated strategy was applied to comprehensively characterize complex multi-type components in Gegen-Qinlian Decoction. A total of 381 compounds were characterized and annotated with four different confidence levels, and 88.40% of these annotated compounds were successfully re-identified in triplicate analyses with a different instrument. The integrated strategy was demonstrated powerful in deep profiling of chemicalome in Chinese medicinal formulas with higher throughput, analytical sharpness, and lower omission ratios.


Assuntos
Medicamentos de Ervas Chinesas , Espectrometria de Massas em Tandem , China , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Espectrometria de Massas em Tandem/métodos
2.
Analyst ; 147(6): 1236-1244, 2022 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-35225997

RESUMO

Collision cross section (CCS) values generated from ion mobility mass spectrometry (IM-MS) have commonly been employed to facilitate lipid identification. However, this is hindered by the limited available lipid standards. Recently, CCS values were predicted by means of computational calculations, though the prediction precision was generally not good and the predicted CCS values of the lipid isomers were almost identical. To address this challenge, a least absolute shrinkage and selection operator (LASSO)-based prediction method was developed for the prediction of lipids' CCS values in this study. In this method, an array of molecular descriptors were screened and optimized to reflect the subtle differences in structures among the different lipid isomers. The use of molecular descriptors together with a wealth of standard CCS values for the lipids (365 in total) significantly improved the accuracy and precision of the LASSO model. Its accuracy was externally validated with median relative errors (MREs) of <1.1% using an independent data set. This approach was demonstrated to allow differentiation of cis/trans and sn-positional isomers. The results also indicated that the LASSO-based prediction method could practically reduce false-positive identifications in IM-MS-based lipidomics.


Assuntos
Espectrometria de Mobilidade Iônica , Lipidômica , Espectrometria de Mobilidade Iônica/métodos , Isomerismo , Lipídeos/análise
3.
J Pharm Biomed Anal ; 208: 114461, 2022 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-34775190

RESUMO

Liver toxicity induced by Triptolide (TP) has limited its clinical application on rheumatoid arthritis (RA). Saponins have been proved as an efficacious remedy to mitigate hepatotoxicity. However, the mechanism of reducing hepatotoxicity by saponins intervention remains incompletely characterized. Tryptophan (Trp) metabolites activate transcriptional regulators to mediate host detoxification responses. Our study aimed to investigate whether Clematichinenoside AR (C-AR) could attenuate TP-induced liver damage by regulating Trp metabolism. We used targeted metabolomics to quantify Trp metabolites in the serum and liver samples of collagen-induced arthritis rats treated by TP. Multiple comparison analyses helped the evaluation of promising biomarkers. The pronounced changed levels of Trp, indole acetic acid, and indole-3-carboxaldehyde in the serum and indole acetic acid, indole, and tryptamine in the liver are relevant to TP-induced liver injury. Intervention with C-AR could relieve TP-induced hepatotoxicity evidenced by ameliorative serum parameters and hepatic histology. In addition, C-AR regulated the levels of these indoles biomarker candidates to normal. Therapeutic modulation with natural compounds might be a useful clinical strategy to ameliorate toxicity induced by TP. Deciphering Trp metabolism will facilitate a better understanding of the pathogenesis of diseases and drug responding.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Diterpenos , Fenantrenos , Saponinas , Animais , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Diterpenos/toxicidade , Compostos de Epóxi/toxicidade , Fígado , Fenantrenos/toxicidade , Ratos , Triterpenos , Triptofano
4.
Mater Today Bio ; 12: 100157, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34825161

RESUMO

Drug-induced liver injury (DILI) is a challenging clinical problem with respect to both diagnosis and management. As a newly emerging biomarker of liver injury, miR122 shows great potential in early and sensitive in situ detection of DILI. Glycyrrhetinic acid (GA) possesses desirable therapeutic effect on DILI, but its certain dose-dependent side effects after long-term and/or high-dose administration limit its clinical application. In this study, in order to improve the precise diagnosis and effective treatment of DILI, GA loaded all-in-one theranostic nanoplatform was designed by assembling of upconversion nanoparticles and gold nanocages. As a proof of concept, we demonstrated the applicability of this single-wavelength laser-triggered theranostic nanoplatform for the spatiotemporally controllable in situ imaging of DILI and miR122-controlled on-demand drug release in vitro and in vivo. This novel nanoplatform opens a promising avenue for the clinical diagnosis and treatment of DILI.

5.
J Med Chem ; 64(20): 14942-14954, 2021 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-34644502

RESUMO

Icaritin is an active ingredient in Epimedium, which has a variety of pharmacological activities. However, the low activity of Icaritin and the unclear target greatly limit its application. Therefore, based on the structure of Icaritin, we adopted the strategy of replacing toxic groups and introducing active groups to design and synthesize a series of new analogues. The top compound C3 exhibited better antimultiple myeloma activity with an IC50 of 1.09 µM for RPMI 8226 cells, induced RPMI 8226 apoptosis, and blocked the cell cycle in the S phase. Importantly, transcriptome analysis, cellular thermal shift assay, and microscale thermophoresis assay confirmed that DEPTOR was the target of C3. Moreover, we explored its binding mode with C3. Especially, C3 displayed satisfactory inhibition of tumor growth in RPMI 8226 xenografts without obvious side effects. In summary, C3 was discovered as a novel putative inhibitor of DEPTOR for the treatment of multiple myeloma.


Assuntos
Antineoplásicos/farmacologia , Desenho de Fármacos , Flavonoides/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Mieloma Múltiplo/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Epimedium/química , Flavonoides/síntese química , Flavonoides/química , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Estrutura Molecular , Mieloma Múltiplo/metabolismo , Relação Estrutura-Atividade
6.
Food Funct ; 12(17): 7607-7618, 2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34236368

RESUMO

Diabetic nephropathy (DN) is a microvascular complication that is becoming a worldwide public health concern. The aim of this study was to assess the effects of dietary soy isoflavone intervention on renal function and metabolic syndrome markers in DN patients. Seven databases including Medline, the Cochrane Central Register of Controlled Trials, Science Direct, Web of Science, Embase, China National Knowledge Infrastructure, and WanFang were searched for controlled trials that assessed the effects of soy isoflavone treatment in DN patients. Finally, a total of 141 patients from 7 randomized controlled trials were included. The meta-analysis showed that dietary soy isoflavones significantly decreased 24-hour urine protein, C-reactive protein (CRP), blood urea nitrogen (BUN), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and fasting blood glucose (FBG) in DN patients. The standard mean difference was -2.58 (95% CI: -3.94, -1.22; P = 0.0002) for 24-hour urine protein, -0.67 (95% CI: -0.94, -0.41; P < 0.00001) for BUN, -6.16 (95% CI: -9.02, -3.31; P < 0.0001) for CRP, -0.58 (95% CI: -0.83, -0.33; P < 0.00001) for TC, -0.41 (95% CI: -0.66, -0.16; P < 0.00001) for TG, -0.68 (95% CI: -0.94, -0.42; P < 0.00001) for LDL-C, and -0.39 (95% CI: -0.68, -0.10; P = 0.008) for FBG. Therefore, soy isoflavones may ameliorate DN by significantly decreasing 24-hour urine protein, BUN, CRP, TC, TG, LDL-C, and FBG.


Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Isoflavonas/administração & dosagem , Adulto , Idoso , Glicemia/metabolismo , Proteína C-Reativa/metabolismo , LDL-Colesterol/metabolismo , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/fisiopatologia , Suplementos Nutricionais/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ensaios Clínicos Controlados Aleatórios como Assunto , Triglicerídeos/metabolismo
7.
Anal Chem ; 93(24): 8536-8543, 2021 06 22.
Artigo em Inglês | MEDLINE | ID: mdl-34107211

RESUMO

Nitric oxide (NO) is a molecule of physiological importance, and the function of NO depends on its concentration in biological systems, particularly in cells. Concentration-based analysis of intracellular NO can provide insight into its precise role in health and disease. However, current methods for detecting intracellular NO are still inadequate for quantitative analysis. In this study, we report a quantitative mass spectrometry probe approach to measure NO levels in cells. The probe, Amlodipine (AML), comprises a Hantzsch ester group that reacts with NO to form a pyridine, Dehydro Amlodipine (DAM). Quantification of DAM by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) allows specific measurement of intracellular NO levels. Notably, the AML/NO reaction proceeds rapidly (within 1 s), which is favorable for NO detection considering its large diffusivity and short half-life. Meanwhile, studies under simulated physiological conditions revealed that the AML response to NO is proportional and selective. The presented UPLC-MS/MS method showed high sensitivity (LLOQ = 0.24 nM) and low matrix interference (less than 15%) in DAM quantification. Furthermore, the mass spectrometry probe approach was demonstrated by enabling the measurement of endogenous and exogenous NO in cells. Hence, the quantitative UPLC-MS/MS method developed using AML as a probe is expected to be a new method for intracellular NO analysis.


Assuntos
Óxido Nítrico , Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Reprodutibilidade dos Testes
8.
J Chromatogr A ; 1651: 462307, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34161837

RESUMO

The difficulty of traditional Chinese medicine (TCM) researches lies in the complexity of components, metabolites, and bioactivities. For a long time, there has been a lack of connections among the three parts, which is not conducive to the systematic elucidation of TCM effectiveness. To overcome this problem, a classification-based methodology for simplifying TCM researches was refined from literature in the past 10 years (2011-2020). The theoretical basis of this methodology is set theory, and its core concept is classification. Its starting point is that "although TCM may contain hundreds of compounds, the vast majority of these compounds are structurally similar". The methodology is composed by research strategies for components, metabolites and bioactivities of TCM, which are the three main parts of the review. Technical route, key steps and difficulty are introduced in each part. Two perspectives are highlighted in this review: set theory is a theoretical basis for all strategies from a conceptual perspective, and liquid chromatography-mass spectrometry (LC-MS) is a common tool for all strategies from a technical perspective. The significance of these strategies is to simplify complex TCM researches, integrate isolated TCM researches, and build a bridge between traditional medicines and modern medicines. Potential research hotspots in the future, such as discovery of bioactive ingredients from TCM metabolites, are also discussed. The classification-based methodology is a summary of research experience in the past 10 years. We believe it will definitely provide support and reference for the following TCM researches.


Assuntos
Técnicas de Química Analítica/métodos , Cromatografia Líquida , Medicamentos de Ervas Chinesas/química , Espectrometria de Massas , Medicina Tradicional Chinesa/tendências , Técnicas de Química Analítica/tendências , Humanos , Projetos de Pesquisa
9.
Artigo em Inglês | MEDLINE | ID: mdl-34052560

RESUMO

Although Cynomorium songaricum Rupr. polysaccharide (CSP) has been examined for its effects on glucose regulation, its underlying mechanism is still unclear. To address this issue, a MS-based lipidomics strategy was developed to gain a system-level understanding of the mechanism of CSP on improving type 2 diabetes mellitus (T2DM). UPLC-QTOF/MS and multivariate statistical tools were used to identify the alteration of serum metabolites associated with T2DM and responses to CSP treatment. As a result, 35 potential biomarkers were found and identified in serum, amongst which 26 metabolites were regulated to normal like levels after the administration of CSP. By analyzing the metabolic pathways, glycerophospholipid metabolism was suggested to be closely involved. These results indicated that the intake of CSP exhibited promising anti-diabetic activity, largely due to the regulation of phospholipid metabolism, including phosphatidylcholines, lysophosphatydylcholines, phosphtatidylethanolamines and sphingomyelins.


Assuntos
Cynomorium/química , Diabetes Mellitus Tipo 2/metabolismo , Lipidômica , Polissacarídeos/farmacologia , Animais , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Extratos Vegetais/farmacologia
10.
Talanta ; 231: 122399, 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-33965049

RESUMO

Straightforward and accurate measurement of medical biomarkers is of essential importance in clinical diagnostics and treatments. However, the major challenge is the diversity in dynamic range of different biomarkers ranging from pg mL-1 to µg mL-1 in various body fluids and tissues among patients. Here, we develop a mesoporous silica (MS)-mediated controllable electrochemiluminescence (ECL) quenching of immunosensor that allows accurate immunoassays with simplicity, sensitivity and tunable sensing range. MS is employed to enhance the sensitivity and tune ECL quenching to broaden the detection range just by altering luminophore (Ru(bpy)32+) and coreactant (DBAE) concentration without additional modifications. The immunoassay is followed: homogeneous sandwich immunoreaction, magnetic separation, and ECL quenching detection. As a proof-of-concept, simple and sensitive detection of IgG is achieved ranging from pg mL-1 to µg mL-1, and applications of the strategy are extended by the combination of ECL immunosensor with commercial ELISA kit. This study will not only be expected to serve as a new avenue for the assay of physiological and clinical implications of immunological biomarkers, but also benefit a wide range of applications that require a tunable detection range and ultrahigh sensitivity.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Eletroquímicas , Humanos , Imunoensaio , Medições Luminescentes , Fotometria , Dióxido de Silício
11.
Phytochem Anal ; 32(2): 124-128, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31359524

RESUMO

INTRODUCTION: Traditional methods to derive experimentally-generated relative correction factors (RCFs) for the quantitative analysis of herbal multi-components by single marker (QAMS) method require reference standards and multiple validations with different instruments and columns, which hampers high throughput implementation. OBJECTIVES: To effectively reduce the application amounts of raw material and provide higher and more stable accuracy, this study aimed to develop a method to computationally generate RCFs of herbal components. MATERIALS AND METHODS: This strategy included the published data collection, calibration curves screening, computer algorithm-based RCFs generation and accuracy validation. RESULTS: Using the in silico approach, we have successfully produced 133 RCFs for the multi-component quantitative analysis of 63 widely used herbs. CONCLUSION: Compared with conventional RCFs, this in silico method would be a low cost and highly efficient way to produce practical RCFs for the QAMS method.


Assuntos
Medicamentos de Ervas Chinesas , Cromatografia Líquida de Alta Pressão , Simulação por Computador
12.
Anal Chim Acta ; 1139: 68-78, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33190711

RESUMO

Neurotransmitter (NT) abnormalities in the enteric nervous system have been reported as crucial roles to regulate the intestinal inflammation and gut immune homeostasis. Capturing quantitative changes at the NT metabolome provides an opportunity to develop an understanding of neuroimmune-mediated inflammation. Given the wide diversity of chemical characterizations in the NTs, only partial coverage of the NT metabolome can be simultaneously quantified in a single-run analysis. Herein, we summarized the distribution of functional groups of compound entries in the NT metabolome. Based on this information, an orthogonal dansyl-labeling and label-free dual pretreatment approach was separately designed to target phenol and amine NTs and tertiary amine and choline NTs. By combining the dansyl-labeled and unlabeled NTs within a single vial, a comprehensive and practical approach was optimized for quantifying high coverage of NT metabolome in a single-run analysis on the reversed-phase C18 column. Method validation indicated good linearity with correlation coefficients (R2) > 0.99, intra- and interday accuracy with relative error < ±20%, and precision with relative standard deviations of ≤15%. With this method, we could simultaneously monitor the alterations of cholines, amines, amino acids, tryptophan and phenylalanine biological pathways in dextran sulphate sodium-induced colitis mice. The measured levels of NT metabolome ranged from 0.0007 to 3.540 µg/mg in intestinal contents and 0.013-154.54 µg/mL in serum samples. The NT metabolism was disrupted by colitis, characterized by the changed NT levels in serum and excessive amino acid NTs accumulation in the intestinal contents. We envisage that the orthogonal approach is of great significance for the comprehensive determination of targeted metabolomics. NTs have the potential to be biomarkers for clinical metabolomics.


Assuntos
Sistema Nervoso Entérico , Metabolômica , Animais , Biomarcadores/metabolismo , Sistema Nervoso Entérico/metabolismo , Metaboloma , Camundongos , Neurotransmissores
13.
Anal Chim Acta ; 1136: 187-195, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-33081943

RESUMO

Long chain unsaturated fatty acids (LCUFAs) are emerging as critical contributors to inflammation and its resolution. Sensitive and accurate measurement of LCUFAs in biological samples is thus of great value in disease diagnosis and prognosis. In this work, a fluorous-derivatization approach for UPLC-MS/MS quantification of LCUFAs was developed by employing a pair of fluorous reagents, namely 3-(perfluorooctyl)-propylamine (PFPA) and 2-(perfluorooctyl)-ethylamine (PFEA). With this method, the LCUFAs in biological samples were perfluoroalkylated with PFPA and specifically retained on a fluorous-phase LC column, which largely reduced matrix interferences-induced quantitation deviation. Moreover, PFEA-labeled LCUFAs standards were introduced as one-to-one internal standards to farthest ensure unbiased results. Application of the proposed method enabled a reliable determination of eight typical LCUFAs with high sensitivity (LLOQ ranged from 30 amol to 6.25 fmol) and low matrix interferences (almost less than 10%). Such a high sensitivity could facilitate the determination of small-volume and low-concentration bio-samples. Further metabolic characterization of these targeted LCUFAs was monitored in OVA-induce asthma mice, requiring only 5 µL serum sample. Our results showed that asthmatic attack led to significant disturbances not only in the concentrations but also in the ratio among these LCUFAs. In view of the favorable advantages in sensitivity and accuracy, the present fluorous-paired derivatization approach will be expected to serve as a new avenue for dissecting the physiological and clinical implications of LCUFAs, thereby shedding light on the management of diseases related to their disturbances.


Assuntos
Asma , Espectrometria de Massas em Tandem , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Ácidos Graxos , Ácidos Graxos Insaturados , Camundongos
14.
Nanoscale ; 12(28): 15325-15335, 2020 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-32648877

RESUMO

Drug-induced liver injury (DILI) is increasingly recognized as one of the most challenging global health problems. Conventional in vitro detection methods not only lack specificity and sensitivity but also cannot achieve real-time, straightforward visualization of hepatotoxicity in vivo. Liver-specific miR122 has been observed to be a superior and sensitive biomarker for DILI diagnosis. Herein, a sensitive upconverting nanoprobe synthesized with upconversion nanoparticles (UCNPs) and gold nanorods (GNR) was designed to diagnose hepatotoxicity in vivo. After injection, the nanoprobes accumulated in the liver and were activated by miR122, and the signal amplification technology fully yielded luminescent amplification; hence, the detection sensitivity was improved. Because of the high tissue penetration capability of near-infrared light, this nanoprobe can achieve real-time in situ detection, thereby providing a novel technology for precise biological and medical analysis.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Nanopartículas , Doença Hepática Induzida por Substâncias e Drogas/diagnóstico , Ouro , Humanos , Luminescência , Tecnologia
15.
J Mass Spectrom ; 55(9): e4528, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32559823

RESUMO

Fully understanding the chemicals in an herbal medicine remains a challenging task. Molecular networking (MN) allows to organize tandem mass spectrometry (MS/MS) data in complex samples by mass spectral similarity, which yet suffers from low coverage and accuracy of compound annotation due to the size limitation of available databases and differentiation obstacle of similar chemical scaffolds. In this work, an enhanced MN-based strategy named diagnostic fragmentation-assisted molecular networking coupled with in silico dereplication (DFMN-ISD) was introduced to overcome these obstacles: the rule-based fragmentation patterns provide insights into similar chemical scaffolds, the generated in silico candidates based on metabolic reactions expand the available natural product databases, and the in silico annotation method facilitates the further dereplication of candidates by computing their fragmentation trees. As a case, this approach was applied to globally profile the steroidal alkaloids in Fritillariae bulbus, a commonly used antitussive and expectorant herbal medicine. Consequently, a total of 325 steroidal alkaloids were discovered, including 106 cis-D/E-cevanines, 142 trans-D/E-cevanines, 29 jervines, 23 veratramines, and 25 verazines. And 10 of them were confirmed by available reference standards. Approximately 70% of the putative steroidal alkaloids have never been reported in previous publications, demonstrating the benefit of DFMN-ISD approach for the comprehensive characterization of chemicals in a complex plant organism.


Assuntos
Alcaloides/química , Fritillaria/química , Fitosteróis/química , Plantas Medicinais/química , Espectrometria de Massas em Tandem/métodos , Alcaloides/análise , Simulação por Computador , Estrutura Molecular , Fitosteróis/análise
16.
Plant Mol Biol ; 103(6): 705-718, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32577984

RESUMO

Fritillariae Bulbus are the most commonly used antitussive and edible herbs in China. Based on UPLC-QTOF-MS and UPLC-QQQ-MS, the validated MRM-based non-targeted quantitative method was applied to determinate the contents of 48 Fritillaria alkaloids (FAs) in three Fritillaria species (F. thunbergii Miq., F. unibracteata and F. ussuriensis). The RNA-Seq results showed that gene transcript levels have different expression patterns in three Fritillaria species. Based on transcriptome data, the full-length cDNA sequences of squalene epoxidase gene were cloned and characterized. Natural evolution of squalene epoxidase genes resulted in four mutations (C236R, M489L, G510A and K517R) in three Fritillaria species. Molecular docking analysis showed that the 236 residue is located inside the pocket and the binding center while other three residues are located on the surface of the protein. Functional verification indicated the mutations of SQE (C236R) could effectively increase the activity of SQE and obtain higher yield of 2,3-oxidosqualene in recombinant yeast. And the mutations of SQE (M489L and G510A), which increased the hydrophobicity of the protein surface, could also enhance the activity of SQE. This study provides major insights into the metabolites differentiation of FAs biosynthesis, and a firm foundation for the quality control and metabolic engineering of Fritillariae bulbus.


Assuntos
Fritillaria/enzimologia , Esqualeno Mono-Oxigenase/metabolismo , Alcaloides/metabolismo , Cromatografia Líquida de Alta Pressão , DNA Complementar/genética , Simulação de Acoplamento Molecular , Mutação/genética , Filogenia , Alinhamento de Sequência , Esqualeno Mono-Oxigenase/genética
17.
J Pharm Biomed Anal ; 182: 113118, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32004769

RESUMO

The stems of Dendrobium officinale, a well-known and expensive food material and herbal medicine in Asia, has recently suffered adulterants and counterfeits by using lower-price confusing Dendrobium species such as D. devonianum or D. transparens in the herbal market. However, robust methods that could authenticate D. officinale from its confusing species effectively are still lacking, especially for the dried samples. This study committed to discover specific peptides biomarkers for the authentication of D. officinale from the other two Dendrobium species using label-free proteomics by nanoLC LTQ Orbitrap mass spectrometry. Multivariate statistical analysis was applied to visualize the difference between the three Dendrobium species. As a result, 29 peptides among a total of 343 measurable peptides were selected to be potential biomarkers for the classification of these Dendrobium species. The validation of the representative peptide biomarkers was carried out by the synthesized peptides and 3 peptide biomarkers were found significant for the authentication of D. officinale. Further analysis showed that peptide ALGLELDLSER may also be a biomarker for the discrimination of the D. officinale originated from different geographical regions.


Assuntos
Dendrobium/química , Peptídeos/química , Extratos Vegetais/química , Proteômica/métodos , Biomarcadores/química , Dendrobium/classificação , Peptídeos/isolamento & purificação , Caules de Planta
18.
Anal Chim Acta ; 1095: 118-128, 2020 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-31864612

RESUMO

A novel liquid-liquid-solid membrane microextraction (LLSMME) method which integrates hollow fiber liquid phase microextraction (HF-LPME) and solid phase microextraction (SPME) was developed for bio-sample preparation. The homogeneous zeolitic imidazolate framework 8 mixed matrix membrane (ZIF-8-MMM) was prepared by in situ self-assembly of ZIF-8 on the inner surface of hollow fiber membrane and employed as a flexible LLSMME device. Incorporating the advantages of both HF-LPME and SPME, the as-fabricated ZIF-8-MMM exhibited excellent performance on the extraction and preconcentration of small molecule drugs of different polarity from complex biological matrices. As a case study, ZIF-8-MMM-based LLSMME coupled with UPLC-MS/MS were developed and validated for determination of ibuprofen, simvastatin and ranitidine at trace levels in rat plasma. The method showed good linearity (r2 > 0.99) and repeatability (RSD < 15%), low limits of detection (2-3 ng mL-1) and high relative recoveries (97.42-103.8%). The enrichment factors were between 87.3 and 112.6. Our study provided a promising strategy for developing more efficient, cost-effective and environmentally friendly technique for bio-sample pretreatment.


Assuntos
Ibuprofeno/sangue , Microextração em Fase Líquida/métodos , Ranitidina/sangue , Sinvastatina/sangue , Microextração em Fase Sólida/métodos , Zeolitas/química , Animais , Cromatografia Líquida de Alta Pressão , Ibuprofeno/isolamento & purificação , Imidazóis/química , Limite de Detecção , Membranas Artificiais , Simulação de Acoplamento Molecular , Ranitidina/isolamento & purificação , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sinvastatina/isolamento & purificação , Espectrometria de Massas em Tandem
19.
J Chromatogr A ; 1612: 460630, 2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-31677768

RESUMO

Authentication of original species is embedded in the quality control system of herbal medicines. In this work, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based untargeted metabolomics coupled with chemometric analysis was utilized for the precise authentication of the Fritillaria species for both raw materials and commercial products. First, a stepwise difference-enlarging chemometric analysis strategy was proposed to analyze eight medicinal Fritillaria species. Subsequently, 21 species-specific markers were discovered and the specificity was investigated under different sample preparation methods. Finally, the obtained species-specific markers were successfully utilized to identify the Fritillaria species in commercially relevant products. This work is the first to report robust and specific markers for authentication of Fritillaria products, showing promise for tracking the supply chain of herbal suppliers.


Assuntos
Alcaloides/análise , Fritillaria/química , Esteroides/análise , Cromatografia Líquida , Fritillaria/metabolismo , Espectrometria de Massas , Metabolômica , Plantas Medicinais/química , Plantas Medicinais/metabolismo , Especificidade da Espécie
20.
Pharmacol Res ; 149: 104459, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31541689

RESUMO

Gut microbiota and their metabolites (short-chain fatty acids, SCFAs) are associated with the pathogenesis of rheumatoid arthritis (RA). Total Clematis triterpenoid saponins (CTSs) prepared from Clematis mandshurica Rupr. possess therapeutic benefits for arthritic diseases. However, the poor pharmacokinetic properties of CTSs have obstructed the translation of these natural agents to drugs. Here, we examined the effects of CTSs on arthritis symptoms, gut microbiota and SCFAs in rats with collagen-induced arthritis (CIA). Our results showed that the arthritis index scores of CIA rats treated with CTSs were significantly lower than those of the model group. Most importantly, CTSs moderated gut microbial dysbiosis and significantly downregulated the total SCFA concentration in CIA rats. Compared to the control group, CTSs treatment have no significant side effects on the gut microbiota and SCFA metabolism in normal rats. Two differential analyses (LEfSe and DESeq2) were combined to study the details of the changes in gut microbiome, and twenty-four marker taxa at the genus level were identified via a comparison among control, model and CIA rats treated with high doses of CTSs. In particular, the mostly significantly increased gram-negative (G-) and decreased gram-positive (G+) genera in CIA rats were well restored by CTSs. The observed SCFA concentrations demonstrated that CTSs tend to maintain the balance of the gut microbiota. The data presented herein suggest that CTSs could ameliorate arthritis-associated gut microbial dysbiosis and may be potential adjuvant drugs that could provide relief from the gastrointestinal damage caused as a side effect of commonly used drugs.


Assuntos
Artrite Experimental/tratamento farmacológico , Clematis/química , Disbiose/prevenção & controle , Ácidos Graxos Voláteis/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Saponinas/uso terapêutico , Triterpenos/uso terapêutico , Animais , Artrite Experimental/microbiologia , Disbiose/microbiologia , Feminino , Ratos , Ratos Wistar , Saponinas/isolamento & purificação , Triterpenos/isolamento & purificação
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