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1.
J Proteomics ; 215: 103669, 2020 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-31987925

RESUMO

The selection of a data processing method for use in mass spectrometry-based label-free proteome quantification contributes significantly to its accuracy and precision. In this study, we comprehensively evaluated 7 commonly-used label-free quantification methods (MaxQuant-Spectrum count, MaxQuant-iBAQ, MaxQuant-LFQ, MaxQuant-LFAQ, Proteome Discoverer, MetaMorpheus, TPP-StPeter) with a focus on missing values, precision, accuracy, selectivity, and reproducibility of low abundance protein quantification in both single shot and fractionation. Our results showed that among the tested strategies, MaxQuant in MaxLFQ mode outperformed other strategies in terms of accuracy and precision in both whole proteome and low abundance proteome quantification, whereas the Proteome Discoverer (PD) strategy using SEQUEST as a search engine performed better in terms of quantifiable low abundance proteome coverage. We subsequently applied the PD and MaxLFQ strategies in a blood proteomic dataset and found that many FDA-approved tumor prognostic biomarkers could be identified as well as quantified using the PD strategy, indicating the potential advantage of PD in label-free quantification studies. These results provide a reference for method choice in label-free quantification data analysis. SIGNIFICANCE: Mass spectrometry-based label-free quantification methods play an important role in label-free proteome data analysis. In this study, we evaluated 7 commonly-used label-free quantification methods with respect to the following aspects: missing values, precision, accuracy, selectivity, and reproducibility for low abundance protein quantification. The results showed that, among the strategies evaluated, the PD strategy with SEQUEST as a search engine performed better in terms of low abundance protein coverage. This study provides a reference for method choice in label-free quantification data analysis.

2.
J Proteomics ; 213: 103614, 2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-31846764

RESUMO

Lysine methylation is a widespread protein post-translational modification showing essentialities in versatile cellular process. EZH2, a methyltransferase specifically trimethylates the lysine 27 of histone H3 and its aberrance in several cancers promotes the development of its inhibitors against hematological tumors. In this study, we presented a deep exploration of lysine mono-, di- and trimethylomes in EZH2 wild-type and Y641 mutant lymphoma cell lines. Our results showed that several substrates were modified in different methylation levels. Moreover, these methylated lysine residues could also undergo other types of PTMs. Combined with the differences proved in protein expression, lysine acetylation, lysine ubiquitylation and protein N-termianl acetylation level, our study underlined the substrate specificity of lysine methylation and its crosstalk with other types of PTMs. Totally, our study raised new insights into the global cellular methylation features in hematological cell lines, which provided further inspects into the distribution and function of lysine methylation. SIGNIFICANCE: Our study showed the global landscape of mono-, di- and trimethylomes in the EZH2-aberrant DLBCL cell lines, revealing the molecular characteristics of lysine methylation. Combined with the protein abundance and potential crosstalk among different types of PTMs, our study raised new insights into the global cellular methylation features in hematological tumors and provided further inspects into the distribution and function of lysine methylation.

3.
Anal Chem ; 91(22): 14522-14529, 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31634432

RESUMO

Global identification of protein C-termini is highly challenging due to their low abundance in conventional shotgun proteomics. Several enrichment strategies have been developed to facilitate the detection of C-terminal peptides. One major issue of previous approaches is the limited C-terminome coverage. Herein, we integrated LysargiNase digestion, chemical acetylation on neo-N-terminus, and a-ion-aided peptide matching into poly(allylamine)-based C-terminomics (termed as LAACTer). In this strategy, we leveraged LysargiNase, a protease with cleavage specificity N-terminal to Lys and Arg residues, to cover previously unidentifiable C-terminome and employed chemical acetylation and a-ion-aided peptide matching to efficiently boost peptide identifications. Triplicates of LAACTer identified a total of 834 C-termini from proteome of 293T cell, which expanded the coverage by 164% (643 more unique C-termini) compared with the parallel experiments using the original workflow. Compared with the largest human C-terminome data sets (containing 800-900 C-termini), LAACTer not only achieved comparable profiling depth but also yielded 465 previously unidentified C-termini. In a SILAC (stable isotope labeling with amino acids in cell culture)-based quantitative study for identification of GluC-cleaved products, LAACTer quantified 300% more C-terminal peptides than the original workflow. Using LAACTer and the original workflow, we performed global analysis for the C-terminal sequences of 293T cell. The original and processed C-termini displayed distinct sequence patterns, implying the "C-end rules" that regulates protein stability could be more complex than just amino acid motifs. In conclusion, we reason LAACTer could be a powerful proteomic tool for in-depth C-terminomics and would benefit better functional understanding of protein C-termini.

4.
Cell Chem Biol ; 25(8): 984-995.e6, 2018 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-29887264

RESUMO

Coenzyme A (CoA) esters of short fatty acids (acyl-CoAs) function as key precursors for the biosynthesis of various natural products and the dominant donors for lysine acylation. Herein, we investigated the functional interplay between beneficial and adverse effects of acyl-CoA supplements on the production of acyl-CoA-derived natural products in microorganisms by using erythromycin-biosynthesized Saccharopolyspora erythraea as a model: accumulation of propionyl-CoA benefited erythromycin biosynthesis, but lysine propionylation inhibited the activities of important enzymes involved in biosynthetic pathways of erythromycin. The results showed that the overexpression of NAD+-dependent deacylase could circumvent the inhibitory effects of high acyl-CoA concentrations. In addition, we demonstrated the similar lysine acylation mechanism in other acyl-CoA-derived natural product biosynthesis, such as malonyl-CoA-derived alkaloid and butyryl-CoA-derived bioalcohol. These observations systematically uncovered the important role of protein acylation on interaction between the accumulation of high concentrations of acyl-CoAs and the efficiency of their use in metabolic pathways.


Assuntos
Acil Coenzima A/metabolismo , Produtos Biológicos/metabolismo , Vias Biossintéticas , Eritromicina/metabolismo , Saccharopolyspora/enzimologia , Saccharopolyspora/metabolismo , Acilação , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Lisina/metabolismo , Processamento de Proteína Pós-Traducional , Saccharopolyspora/química , Metabolismo Secundário
5.
ACS Chem Biol ; 13(6): 1588-1597, 2018 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-29799716

RESUMO

Protein acylation plays important roles in bacterial pathogenesis through regulation of enzymatic activity, protein stability, nucleic acid binding ability, and protein-protein interactions. Mycobacteria, a genus including invasive pathogens known to cause serious diseases, shapes its pathogenicity through adaptation of its energy metabolism to microenvironments encountered within mammalian hosts. In this process, acetyl-CoA and propionyl-CoA function as important intermediates. However, the function of acetyl-CoA/propionyl-CoA driven protein acylation remains to be elucidated. Herein, we systematically investigated protein acetylome/propionylome in the nonpathogenic Mycobacterium smegmatis through antibody-enrichment-based proteomic analysis in which 146 acetylated sites on 121 proteins and 26 propionylated sites on 25 proteins were identified. After that, characteristic differences of the two acylomes were elucidated through such bioinformatic methods as motif analysis, protein-protein analysis, Gene Ontology analysis, and KEGG analysis. In addition, quantitative mass spectrometric method was used to evaluate the site-specific and motif-biased catalytic mechanism mediated by the cAMP-dependent acetyltransferase MsKat in M. smegmatis. Furthermore, we raised the possibility that both O-serine and Nε-lysine acetylation might coregulate the propionyl-CoA synthetase. This study described the landscape of acetylome and propionylome in the M. smegmatis, showing an unexpected role of protein acylation regulation in mycobacteria.


Assuntos
Acetiltransferases/metabolismo , Proteínas de Bactérias/análise , Lisina/química , Mycobacterium smegmatis/enzimologia , Proteoma/análise , Acetilação , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Biologia Computacional/métodos , Cinética , Espectrometria de Massas/métodos , Processamento de Proteína Pós-Traducional , Proteoma/química , Proteoma/metabolismo , Proteômica/métodos
6.
ACS Chem Biol ; 13(5): 1200-1208, 2018 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-29690763

RESUMO

The effect of regulatory system on the engineered biosynthetic pathway in chassis cells remains incompletely understood in microorganisms. Acyl-CoAs function as key precursors for the biosynthesis of various natural products and the dominant donors for protein acylation. The polyphenol pinosylvin, with high antimicrobial and antifungal activities, is biosynthesized with malonyl-CoA as its direct precursors. But correlation between lysine malonylation and pinosylvin biosynthesis remains unknown. Herein, we found that the malonyl-CoA-driven lysine malonylation plays an important role in interaction between the engineered pathway of pinosylvin synthesis and E. coli chassis cell. Oversupply of malonyl-CoA leads to an increase in malonylation level of global proteome as well as the enzymes in the artificial pathway, thereby decreasing yield of pinosylvin. The results revealed that the intricate balance of cellular acyl-CoA concentrations is critical for the yields of acyl-CoA-derived natural products. We next modified the enzymes in the biosynthetic pathway to adjust their acylation level and successfully improved the yield of pinosylvin. Our study uncovers the effect of protein acylation on the biosynthetic pathway, helps optimization of synthetic constructs, and provides new strategies in metabolic engineering and synthetic biology at the protein post-translational level.


Assuntos
Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Engenharia Metabólica , Estilbenos/metabolismo , Acilação , Vias Biossintéticas , Escherichia coli/genética , Processamento de Proteína Pós-Traducional
7.
Mol Cell Proteomics ; 17(6): 1156-1169, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29523768

RESUMO

Clostridium acetobutylicum is a strict anaerobic, endospore-forming bacterium, which is used for the production of the high energy biofuel butanol in metabolic engineering. The life cycle of C. acetobutylicum can be divided into two phases, with acetic and butyric acids being produced in the exponential phase (acidogenesis) and butanol formed in the stationary phase (solventogenesis). During the transitional phase from acidogenesis to solventogenesis and latter stationary phase, concentration peaks of the metabolic intermediates butyryl phosphate and acetyl phosphate are observed. As an acyl group donor, acyl-phosphate chemically acylates protein substrates. However, the regulatory mechanism of lysine acetylation and butyrylation involved in the phenotype and solventogenesis of C. acetobutylicum remains unknown. In our study, we conducted quantitative analysis of protein acetylome and butyrylome to explore the dynamic change of lysine acetylation and butyrylation in the exponential phase, transitional phase, and stationary phase of C. acetobutylicum Total 458 lysine acetylation sites and 1078 lysine butyrylation sites were identified in 254 and 373 substrates, respectively. Bioinformatics analysis uncovered the similarities and differences between the two acylation modifications in C. acetobutylicum Mutation analysis of butyrate kinase and the central transcriptional factor Spo0A was performed to characterize the unique role of lysine butyrylation in the metabolic pathway and sporulation process of C. acetobutylicum Moreover, quantitative proteomic assays were performed to reveal the relationship between protein features (e.g. gene expression level and lysine acylation level) and metabolites in the three growth stages. This study expanded our knowledge of lysine acetylation and butyrylation in Clostridia and constituted a resource for functional studies on lysine acylation in bacteria.


Assuntos
Proteínas de Bactérias/metabolismo , Butiratos/metabolismo , Clostridium acetobutylicum/metabolismo , Acetilação , Lisina/metabolismo , Redes e Vias Metabólicas , Fenótipo , Fosfotransferases (Aceptor do Grupo Carboxila)/genética , Esporos Bacterianos , Fatores de Transcrição/genética
8.
Neurosci Bull ; 34(2): 237-246, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28936771

RESUMO

N-methyl-D-aspartate receptors (NMDARs), a subtype of glutamate-gated ion channels, play a central role in epileptogenesis. Recent studies have identified an increasing number of GRIN2A (a gene encoding the NMDAR GluN2A subunit) mutations in patients with epilepsy. Phenotypes of GRIN2A mutations include epilepsy-aphasia disorders and other epileptic encephalopathies, which pose challenges in clinical treatment. Here we identified a heterozygous GRIN2A mutation (c.1341T>A, p.N447K) from a boy with Rolandic epilepsy by whole-exome sequencing. The patient became seizure-free with a combination of valproate and lamotrigine. Functional investigation was carried out using recombinant NMDARs containing a GluN2A-N447K mutant that is located in the ligand-binding domain of the GluN2A subunit. Whole-cell current recordings in HEK 293T cells revealed that the N447K mutation increased the NMDAR current density by ~1.2-fold, enhanced the glutamate potency by 2-fold, and reduced the sensitivity to Mg2+ inhibition. These results indicated that N447K is a gain-of-function mutation. Interestingly, alternative substitutions by alanine and glutamic acid at the same residue (N447A and N447E) did not change NMDAR function, suggesting a residual dependence of this mutation in altering NMDAR function. Taken together, this study identified human GluN2A N447K as a novel mutation associated with epilepsy and validated its functional consequences in vitro. Identification of this mutation is also helpful for advancing our understanding of the role of NMDARs in epilepsy and provides new insights for precision therapeutics in epilepsy.


Assuntos
Epilepsia Rolândica/genética , Receptores de N-Metil-D-Aspartato/genética , Adolescente , Humanos , Masculino , Mutação
9.
Proteomics ; 18(1)2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29150981

RESUMO

The studies of protein methylation mainly focus on lysine and arginine residues due to their diverse roles in essential cellular processes from gene expression to signal transduction. Nevertheless, atypical protein methylation occurring on amino acid residues, such as glutamine and glutamic acid, is largely neglected until recently. In addition, the systematic analysis for the distribution of methylation on different amino acids in various species is still lacking, which hinders our understanding of its functional roles. In this study, we deeply explored the methylated sites in three species Escherichia coli, Saccharomyces cerevisiae, and HeLa cells by employing MS-based proteomic approach coupled with heavy methyl SILAC method. We identify a total of 234 methylated sites on 187 proteins with high localization confidence, including 94 unreported methylated sites on nine different amino acid residues. KEGG and gene ontology analysis show the pathways enriched with methylated proteins are mainly involved in central metabolism for E. coli and S. cerevisiae, but related to spliceosome for HeLa cells. The analysis of methylation preference on different amino acids is conducted in three species. Protein N-terminal methylation is dominant in E. coli while methylated lysines and arginines are widely identified in S. cerevisiae and HeLa cells, respectively. To study whether some atypical protein methylation has biological relevance in the pathological process in mammalian cells, we focus on histone methylation in diet-induced obese (DIO) mouse. Two glutamate methylation sites showed statistical significance in DIO mice compared with chow-fed mice, suggesting their potential roles in diabetes and obesity. Together, these findings expanded the methylome database from microbes to mammals, which will benefit our further appreciation for the protein methylation as well as its possible functions on disease.


Assuntos
Aminoácidos/metabolismo , Escherichia coli/metabolismo , Histonas/metabolismo , Obesidade/metabolismo , Processamento de Proteína Pós-Traducional , Saccharomyces cerevisiae/metabolismo , Aminoácidos/química , Animais , Bases de Dados Factuais , Evolução Molecular , Células HeLa , Histonas/química , Humanos , Masculino , Metilação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Obesidade/etiologia , Proteômica , Especificidade por Substrato
10.
J Proteome Res ; 15(5): 1685-701, 2016 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-27090497

RESUMO

Lysine acylation is a dynamic, reversible post-translational modification that can regulate cellular and organismal metabolism in bacteria. Acetylome has been studied well in bacteria. However, to our knowledge, there are no proteomic data on the lysine malonylation in prokaryotes, especially in actinomycetes, which are the major producers of therapeutic antibiotics. In our study, the first malonylome of the erythromycin-producing Saccharopolyspora erythraea was described by using a high-resolution mass spectrometry-based proteomics approach and high-affinity antimalonyllysine antibodies. We identified 192 malonylated sites on 132 substrates. Malonylated proteins are enriched in many biological processes such as protein synthesis, glycolysis and gluconeogenesis, the TCA cycle, and the feeder metabolic pathways of erythromycin synthesis according to GO analysis and KEGG pathway analysis. A total of 238 S/T/Y/H-phosphorylated sites on 158 proteins were also identified in our study, which aimed to explore the potential cross-talk between acylation and phosphorylation. After that, site-specific mutations showed that malonylation is a negative regulatory modification on the enzymatic activity of the acetyl-CoA synthetase (Acs) and glutamine synthetase (Gs). Furthermore, we compared the malonylation levels of the two-growth state to explore the potential effect of malonylation on the erythromycin biosynthesis. These findings expand our current knowledge of the actinomycetes malonylome and supplement the acylproteome databases of the whole bacteria.


Assuntos
Eritromicina/biossíntese , Lisina/metabolismo , Malonatos/metabolismo , Saccharopolyspora/metabolismo , Vias Biossintéticas , Metabolismo , Processamento de Proteína Pós-Traducional , Proteômica/métodos
11.
J Biol Chem ; 289(39): 27034-45, 2014 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-25124041

RESUMO

ACT domains (amino acid-binding domains) are linked to a wide range of metabolic enzymes that are regulated by amino acid concentration. Seventy proteins with ACT-GCN5-related N-acetyltransferase (GNAT) domain organization were found in actinomycetales. In this study, we investigate the ACT-containing GNAT acetyltransferase, Micau_1670 (MaKat), from Micromonospora aurantiaca ATCC 27029. Arginine and cysteine were identified as ligands by monitoring the conformational changes that occur upon amino acids binding to the ACT domain in the MaKat protein using FRET assay. It was found that MaKat is an amino acid-regulated protein acetyltransferase, whereas arginine and cysteine stimulated the activity of MaKat with regard to acetylation of acetyl-CoA synthetase (Micau_0428). Our research reveals the biochemical characterization of a protein acetyltransferase that contains a fusion of a GNAT domain with an ACT domain and provides a novel signaling pathway for regulating cellular protein acetylation. These findings indicate that acetylation of proteins and acetyltransferase activity may be tightly linked to cellular concentrations of some amino acids in actinomycetales.


Assuntos
Acetiltransferases/metabolismo , Arginina/metabolismo , Proteínas de Bactérias/metabolismo , Cisteína/metabolismo , Micromonospora/enzimologia , Acetilação , Acetiltransferases/genética , Regulação Alostérica/fisiologia , Arginina/genética , Proteínas de Bactérias/genética , Coenzima A Ligases/genética , Coenzima A Ligases/metabolismo , Cisteína/genética , Micromonospora/genética
12.
Int J Clin Pharmacol Ther ; 51(7): 568-75, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23611568

RESUMO

OBJECTIVES: The aim of this study is to investigate the population pharmacokinetics (PopPK) of cyclosporine (CsA) in the Chinese hematopoietic stem cell transplantation (HSCT) recipients for promoting the individualization of CsA administration. METHODS: A total of 887 retrospective drug monitoring data points were collected from 58 HSCT recipients. Whole blood samples were collected at predose (C0) and 2 hours (C2) post dose. The administration of CsA was intermittent intravenous infusion, continuous intravenous infusion and oral. Population modeling was performed using the NONMEM (nonlinear mixedeffect modeling) program. A one compartment pharmacokinetic model was used to fit the data. RESULTS: Body surface area (BSA), administration route and postoperative days were identified as significant covariates for clearance (CL) according to the final model: CL = 31.0 × (BSA/1.59)0.761 × (ROUT) × (POD), where ROUT was 1.91 if the administration route was intravenous infusion, otherwise it is equal to 1. The POD was 0.818, 0.753, 0.539, and 0.509 for posttransplant Days 0 - 10, 11 - 20, 21 - 30 and more than 30 days, respectively. Administration route was a significant covariate for volume (V) according to the final model: V = 192 × (ROUT), where ROUT was 4.10, 3.63 and 1 when the administration route was continuous intravenous infusion, intermittent intravenous infusion and oral. The other covariates were not identified as a significant effect on CsA pharmacokinetic parameters. CONCLUSION: Body surface area, administration route and postoperative days should be considered in individual pharmacotherapy of cyclosporine for HSCT patient to achieve the desired therapeutic target.


Assuntos
Ciclosporina/farmacocinética , Transplante de Células-Tronco Hematopoéticas , Imunossupressores/farmacocinética , Administração Oral , Adolescente , Adulto , Grupo com Ancestrais do Continente Asiático , Superfície Corporal , Criança , Pré-Escolar , China , Ciclosporina/administração & dosagem , Ciclosporina/sangue , Esquema de Medicação , Cálculos da Dosagem de Medicamento , Monitoramento de Medicamentos , Transplante de Células-Tronco Hematopoéticas/etnologia , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/sangue , Infusões Intravenosas , Taxa de Depuração Metabólica , Pessoa de Meia-Idade , Modelos Biológicos , Modelos Estatísticos , Dinâmica não Linear , Estudos Retrospectivos , Adulto Jovem
13.
Sheng Li Xue Bao ; 64(5): 550-62, 2012 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-23090496

RESUMO

Neuroligins (NLs) are postsynaptic membrane proteins expressed in the brain and mediate synaptogenesis. Neuroligin family proteins can specifically induce either excitatory or inhibitory synapses. Deletions or point mutations in neuroligin genes are found in patients with autism spectrum disorders (ASD) or mental retardations. The dysfunctions of these mutations have been tested in multiple neuroligin mouse models. In most of the models, including the human autism-linked NL3 and NL4 mutation mice, there are social interaction defects, memory impairment and repetitive behaviors. Researchers also found the excitatory/inhibitory synapse ratio altered in those mice, as well as receptor subunit composition. However, inconsistencies and debates also exist between different research approaches. In this review, we summarize the neuroligin mouse models currently available, examine the detailed alterations detected in those mice and compare the differences within different mouse models or different investigation methods, to obtain an overall picture of the current progress on neuroligin mouse models.


Assuntos
Encéfalo/fisiopatologia , Moléculas de Adesão Celular Neuronais/fisiologia , Modelos Animais de Doenças , Sinapses/fisiologia , Animais , Transtorno Autístico/fisiopatologia , Humanos , Proteínas de Membrana/fisiologia , Camundongos , Mutação , Proteínas do Tecido Nervoso/fisiologia
14.
Zhongguo Zhong Yao Za Zhi ; 32(24): 2613-9, 2007 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-18338600

RESUMO

OBJECTIVE: To develop a urine pretreatment method of Solid Phase Extraction (SPE) for the quantitative determination of a number of aristolochic acids (AAs) and aristololactams (ALs) in rat urine. METHOD: The HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other sixteen AAs and ALs was chosen as evaluating index to study the extract results of five Solid Phase Extraction columns (Agilent C18/100 mg, Alltech HG18/100 mg, Alltech C18/100 mg, Alltech C18/300 mg and Agilent Phenyl/200 mg) comparatively. The influences of two washing solvents (water and 1% acetic acid-0.02% triethylamine solution) and seven eluting solvents (ether, acetone, chloroform, ethyl acetate, dichloromethane, methanol and acetonitrile) on extract results of AAs and ALs are comparatively studied with the extracting recoveries of AA-I , AA-II, AL-I and AL-II as indicators. The HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other seven AAs and ALs with good separation being targets, several factors which affect extracting efficiency of analytes, including activating volume, cleansing volume, washing volume and eluting volume, are optimized by orthogonal design experiments with four factors at three levels. RESULT: The established method of SPE is as follows: Agilent Phenyl SPE column of 200 mg, activating with 1.0 mL methanol, cleansing with 1 mL water, adding 1.0 mL rat urine sample, washing with 0.8 mL 1% acetic acid 0.02% triethylamine solution, and eluting with 3.0 mL methanol. CONCLUSION: The established method of SPE is efficient, selective, simple and fast, and can be used as urine pretreatment method to analyze a variety of aristolochic acids and aristololactams in rat urine.


Assuntos
Aristolochia , Ácidos Aristolóquicos/urina , Medicamentos de Ervas Chinesas/farmacocinética , Administração Oral , Animais , Aristolochia/química , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/isolamento & purificação , Masculino , Distribuição Aleatória , Ratos , Ratos Wistar , Extração em Fase Sólida/métodos
15.
Zhongguo Zhong Yao Za Zhi ; 30(11): 835-9, 2005 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-16110865

RESUMO

OBJECTIVE: Taking Caulis Aristolochiae Manshuriensis (Guanmutong in Chinese, derived from the stem of Aristolochia manshuriensis) as an example, to study the affection of different preparations on the content of toxic constituents in traditional Chinese medicines. METHOD: The separation was performed on a zorbax SB-C18 column with mobile phase of acetonitrile-3.7 mmol x L(-1) phosphoric acid buffer, detected at 260 nm. RESULT: The extraction percentage of aristolochic acids I, II and IV a in water extraction (1 h x 2) of Guanmutong were 53.4%, 75.5% and 61.9%, respectively; the remaining quantity of aristolochic acids I, II and IVa in the dregs of the decoction were 22.3%, 15.7% and 30.3%, respectively; Aristolochic acid I was still main substance among these aristolohic acids in the decoction of Guanmutong. CONCLUSION: The content of toxic constituents of the traditional Chinese medicines varies evidently with different preparations of Guanmutong. So the preparation methods of traditional Chinese medicines should be suitably selected according to characteristics of the toxic constituents so as to lessen the body damages of human.


Assuntos
Aristolochia/química , Ácidos Aristolóquicos/análise , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Plantas Medicinais/química , Resíduos de Drogas/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Soluções Farmacêuticas/química , Pós/química
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