RESUMEN

Background: Lupus nephritis (LN), caused by Systemic lupus erythematosus, is a chronic autoimmune renal disease and a key risk factor for morbidity and fatality involving 50% damage to the kidney. LN is associated with aberrant functioning of the immune system, characterized by increased systemic inflammation, altered lymphocyte count, perivascular infiltration of inflammatory cells, and declined organ functioning. Current Therapies and Limitations: Conventional therapies to LN include high-dose glucocorticoids, immunosuppressants, calcineurin inhibitors, immune boosters, and targeted medicines that improve kidney functioning. However, these drugs triggered severe adverse side effects, and their prolonged usage resulted in drug resistance, accentuating LN complications. TCM in LN Treatment: Hence, safe and functional Traditional Chinese Medicines (TCM), with supporting clinical trials and observational studies, received significant recognition worldwide for the Treatment of LN. In the form of herbal extracts and preparations, TCM proved effective in treating immunodeficient disorders, including LN. Additionally, acupuncture as a TCM appeared promising in reducing LN-induced inflammation and joint pain. Mechanisms and Benefits: The therapeutic mechanisms included reduced antibody generation, pro-inflammatory cytokine release, immune complex formation, complement activation, extracellular matrix damage and proteinuria levels that played vital roles in chronic kidney diseases. They generated immunosuppressive effects by modulating apoptosis, oxidative stress, and inflammatory signaling pathways, such as JAK/STAT, NF-κB, AP-1 and MAPK and their cross-talk in LN and associated renal injury. These therapies improved blood circulation, alleviated renal pathological changes, restored glomerular capillary functioning, and regenerated renal tissues. However, an essential requisite for these therapies for LN included reduced side effects and improved hepatoprotection and detoxification. Clinical studies suggest that TCM formulations may demonstrate therapeutic benefits in alleviating the symptoms of LN, suggesting prospects of combined applications with Western medicine to enhance treatment efficiencies. Overall, TCM is beneficial for treating LN, and may serve as a potential alternative to conventional medicines.

RESUMEN

A novel rotavirus A (RVA) strain (JZ) was detected in RVA-positive stool specimens from three pediatric patients in Jinzhou, Liaoning Province, in 2018-2019. The electrophoresis pattern of the JZ strain showed a long electropherotype. The genomic constellation G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1 was detected, suggesting that a new inter-genogroup reassortment had occurred. Whole-genome sequencing showed that the JZ isolates were identical to each other. Phylogenetic analysis revealed that VP7 and VP4 clustered in lineages G9-VI and P[8]-3, respectively. JZ strain-specific amino acid substitutions were detected in VP7, VP4 and NSP4. This study provides information on the epidemiology and characteristics of G9 strains circulating in China.

Asunto(s)

Virus Reordenados/aislamiento & purificación , Infecciones por Rotavirus/virología , Rotavirus/genética , Rotavirus/aislamiento & purificación , China , Genoma Viral , Humanos , Filogenia , Virus Reordenados/clasificación , Virus Reordenados/genética , Rotavirus/clasificación , Proteínas Virales/genética

RESUMEN

BACKGROUND: Nelson Bay orthoreovirus (NBV) was first isolated over 40 years ago from a fruit bat in Australia. Normally, NBV does not cause human diseases, but recently several NBV strains have been associated with human respiratory tract infections, thus attracting clinical attention. Autophagy, an evolutionarily conserved process in eukaryotic cells, degrades intracellular substrates, participates in multiple physiological processes, and maintains cellular homeostasis. In addition, autophagy is intimately involved in viral infection. METHODS: A new strain of NBV, isolated from a patient with a respiratory tract infection who returned to Japan from Bali, Indonesia, in 2007, was used in this study. NBV was rescued using a reverse genetics system involving cotransfection of BHK cells with 11 plasmids (pT7-L1 MB, pT7-L2 MB, pT7-L3 MB, pT7-M1 MB, pT7-M2 MB, pT7-M3 MB, pT7-S1 MB, pT7-S2 MB, pT7-S3 MB, pT7-S4 MB, and pcDNA3.1-T7), yielding NBV-MB. Recovered viruses were confirmed by immunofluorescence. The effect of NBV-MB on autophagy was evaluated by measuring the LC3-I/II proteins by immunoblot analysis after infection of BHK cells. Furthermore, after treatment with rapamycin (RAPA), 3-methyladenine (3-MA), chloroquine (CQ), or plasmid (GFP-LC3) transfection, the changes in expression of the LC3 gene and the amount of LC3-I/II protein were examined. In addition, variations in viral titer were assayed after treatment of BHK cells with drugs or after transfection with plasmids pCAGM3 and pCAGS3, which encode virus nonstructural proteins µNS and σNS, respectively. RESULTS: NBV-MB infection induced autophagy in host cells; however, the level of induction was dependent on viral replication. Induction of autophagy increased viral replication. By contrast, inhibiting autophagy suppressed NBV replication, albeit not significantly. The NBV-MB nonstructural protein µNS was involved in the induction of autophagy with viral infection. CONCLUSIONS: NBV-MB infection triggered autophagy. Also, the NBV nonstructural protein µNS may contribute to augmentation of autophagy upon viral infection.

Asunto(s)

Autofagia , Interacciones Microbiota-Huesped , Orthoreovirus/fisiología , Replicación Viral , Línea Celular , Células HEK293 , Humanos , Infecciones por Reoviridae/virología , Genética Inversa , Carga Viral , Proteínas Virales/genética

RESUMEN

Objective To prepare a polyclonal antibody against S3 of Nelson Bay virus (NBV). Methods The E.coli BL21 (DE3) competent cells were transfected with the constructed Pris His MB-S3 recombinant plasmid. Protein expression was induced by IPTG. The target protein was purified by Ni-NTA column affinity chromatography to obtain a large amount of fusion recombinant protein, which was tested and used as the antigen for producing the S3 polyclonal antibody in rats. Results The relative molecular mass (Mr) of Pris His MB-S3 protein was around 39 000, in the form of inclusion bodies. NBV S3 polyclonal antibody was successfully produced from the immunized rats. The titer of the antibody was up to 1:64 000 determined by indirect ELISA. The indirect immunofluorescence assay verified that the S3 protein was successfully expressed in cells and distributed in a granular form in the cytoplasm. Conclusion The highly reactive and specific S3 protein polyclonal antibody is successfully prepared.

Asunto(s)

Anticuerpos , Proteínas de la Cápside/aislamiento & purificación , Orthoreovirus , Proteínas de Unión al ARN/aislamiento & purificación , Animales , Especificidad de Anticuerpos , Proteínas de la Cápside/inmunología , Ensayo de Inmunoadsorción Enzimática , Plásmidos , Proteínas de Unión al ARN/inmunología , Ratas , Proteínas Recombinantes de Fusión

RESUMEN

The present study was designed to develop and validate a rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of five major active constituents in the traditional Chinese medicinal preparation Xingxiong injection (XXI) in rat plasma, including quercetin 3-O-rutinoside (QCR), kaempferol 3-O-rutinoside (KFR), isorhamnetin 3-O-rutinoside (ISR), bilobalide (BB), and ligustrazine (LGT). The plasma samples were pretreated by protein precipitation with acetonitrile. The chromatographic separation was achieved on a Waters Symmetry C18 analytical column (2.1 mm × 100 mm, 3.5 µm) with a mobile phase of 0.1% aqueous formic acid (A)-acetonitrile (B). Quantitation of the five bioactive constituents was achieved. Naringin was used as the internal standard (IS). All the calibration curves showed good linearity (r > 0.996) over the concentration range, with the lowest limit of quantification (LLOQ) between 2-18 ng·mL-1. The intra- and inter-day accuracy and precision of the analytes were both within acceptable limits. Moreover, satisfactory extraction recoveries (90.92%-104.03%) were obtained by protein precipitation. The validated method was successfully applied to a pharmacokinetic study of XXI in rats after intravenous administration at three doses. The pharmacokinetic parameters of the five compounds varied in a dose-dependent manner within the tested dosage range. The present study was the first report of pharmacokinetic study for XXI.

Asunto(s)

Bilobálidos/sangre , Cromatografía Líquida de Alta Presión/métodos , Disacáridos/sangre , Medicamentos Herbarios Chinos/análisis , Flavonoides/sangre , Glucósidos/sangre , Quempferoles/sangre , Pirazinas/sangre , Quercetina/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Animales , Bilobálidos/farmacocinética , Disacáridos/farmacocinética , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/farmacocinética , Flavonoides/farmacocinética , Glucósidos/farmacocinética , Quempferoles/farmacocinética , Pirazinas/farmacocinética , Quercetina/sangre , Quercetina/farmacocinética , Ratas , Ratas Sprague-Dawley

RESUMEN

Ardisiae Japonicae Herba is a well-known traditional Chinese medicine for the treatment of bronchitis conjunctivitis, pneumonia, and trauma. In this work, a high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry method was first established for the separation and structural identification of the chemical constituents in Ardisiae Japonicae Herba. A total of 15 compounds including coumarins, flavonoid glycosides, and catechins were identified or tentatively characterized based on their chromatographic behaviors and mass spectral fragmentation and by comparisons with the reference standards. Furthermore, a simple high-performance liquid chromatography with diode array detection method was developed for the simultaneous determination of five major constituents. Results obtained from method validation, including linearity, precision, repeatability, stability, and recovery, showed that the established method was reliable and accurate. Bergenin and quercitrin were found to be the most abundant constituents and could be served as chemical markers for quality control of Ardisiae Japonicae Herba.

Asunto(s)

Ardisia/química , Catequina/aislamiento & purificación , Cumarinas/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Flavonoides/aislamiento & purificación , Glicósidos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Medicina Tradicional China

RESUMEN

Citrus polymethoxyflavones (PMFs) have been of increasing interest due to their extensive biological activities. In the present study, a total of eight PMFs were isolated from the peel of Citrus reticulata 'Chachi' (CRC). They were individually identified as 5-hydroxy-6,7,8,4'-tetramethoxyflavone (1), 5,6,7,3',4'-pentamethoxyflavanone (2), 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone (3), 3,5,6,7,8,3',4'- heptamethoxyflavone (4), 5,6,7,8,3',4'-hexamethoxyflavone (5), 5,6,7,8,4'-pentamethoxyflavone (6), 6,7,8,3',4'-pentamethoxyflavanone (7) and 5,6,7,3',4'-pentamethoxyflavone (8) by nuclear magnetic resonance and mass spectroscopic analysis. 6,7,8,3',4'-Pentamethoxyflavanone was isolated from the peel of CRC for the first time. The content of PMFs was firstly quantified in both the peel of CRC and PMF-rich extract by HPLC analysis. Furthermore, the biological activities of PMF compounds were investigated. 3,5,6,7,8,3',4'-Heptamethoxyflavone demonstrated potent sterol regulatory element-binding proteins inhibition activity and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone exhibited strong antiproliferative activity against tumor cell lines. The isolated PMF compounds could also significantly inhibit NO production and the effect varied mainly depending on the number of methoxy groups.

Asunto(s)

Citrus/química , Flavonas/química , Frutas/química , Animales , Antineoplásicos Fitogénicos/química , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Humanos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Ratones , Estructura Molecular , Óxido Nítrico/química , Células RAW 264.7

RESUMEN

A new compound, methylbergenin (1), was isolated from the whole plants of Ardisia japonica (Thunb.) Bl. (Myrsinaceae), along with eight known compounds: demethoxybergenin (2), bergenin (3), afzelin (4), quercitrin (5), bauerenol (6), bauerenone (7), α-spinasterol (8) and chondrillasterone (9). Compounds 1, 7 and 9 were isolated from A. japonica for the first time. The structure of compound 1 was determined on the basis of NMR (1D and 2D) and mass spectroscopic analysis. The absolute configuration of 1 was assessed by single-crystal X-ray diffraction. Compounds 1, 4, 5, 7 and 9 showed potential inhibitory effects against nitric oxide production in LPS stimulated RAW 264.7 murine macrophages. In addition, Compounds 7 and 9 exhibited cytotoxicity against A549 and HepG-2 cells.

Asunto(s)

Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/farmacología , Ardisia/química , Animales , Benzopiranos/aislamiento & purificación , Cristalografía por Rayos X , Evaluación Preclínica de Medicamentos/métodos , Células Hep G2 , Humanos , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Espectroscopía de Resonancia Magnética , Ratones , Estructura Molecular , Óxido Nítrico/biosíntesis , Estigmasterol/análogos & derivados , Estigmasterol/aislamiento & purificación , Triterpenos/química , Triterpenos/aislamiento & purificación , Triterpenos/farmacología

RESUMEN

Deep eutectic solvents (DESs) are emerging as alternatives to conventional ionic liquids and organic solvents due to their unique advantages. In the present study, the tuneability of DESs as tailor-made solvents to efficiently extract polar and non-polar bioactive compounds from Platycladi Cacumen was detailedly investigated. Totally 12 types of choline chloride-, betaine-, and l-proline-based DESs were synthesized for initial screening, and extraction conditions was optimized by single-factor experiment. Experiments with different DESs and principal components analysis demonstrated that the extractability of both flavonoid glycosides and aglycones was greater with certain designed DESs than conventional solvents. In addition, the water content in DESs led to significantly different extraction yields of flavonoid compounds. The target compounds were recovered from DESs by macroporous resin LX-38 with a satisfactory yield between 77.44% and 98.92%. The knowledge acquired in this study could contribute to further DES application in extraction of bioactive compounds from natural sources.

Asunto(s)

Medicamentos Herbarios Chinos/análisis , Flavonoides/análisis , Glicósidos/análisis , Tecnología Química Verde/métodos , Iridoides/análisis , Solventes/química

RESUMEN

To get a better understanding of the bioactive constituents in Aurantii Fructus Immaturus (AFI) and Aurantii Fructus (AF), in the present study, a comprehensive strategy integrating multiple chromatographic analysis and chemometrics methods was firstly proposed. Based on segmental monitoring, a high-performance liquid chromatography (HPLC)-variable wavelength detection method was established for simultaneous quantification of ten major flavonoids, and the quantitative data were further analyzed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). A strong cation exchange-high performance liquid chromatography (SCX-HPLC) method combined with t-test and one-way analysis of variance (ANOVA) was developed to determine synephrine, the major alkaloid in AFI and AF. The essential oils were analyzed by gas chromatography-mass spectrometry (GC-MS) and further processed by partial least squares discrimination analysis (PLS-DA). The results indicated that the contents of ten flavonoids and synephrine in AFI were significantly higher than those in AF, and significant difference existed in samples from different geographical origins. Also, 9 differential volatile constituents detected could be used as chemical markers for discrimination of AFI and AF. Collectively, the proposed comprehensive analysis might be a well-acceptable strategy to evaluate the quality of traditional citrus herbs.

Asunto(s)

Citrus/química , Flavonoides/análisis , Frutas/química , Sinefrina/análisis , Cromatografía Líquida de Alta Presión , Análisis por Conglomerados , Cromatografía de Gases y Espectrometría de Masas , Aceites Volátiles/química , Aceites de Plantas/química , Análisis de Componente Principal

RESUMEN

Selection of suitable labeled constituents is vital for the quality standardization of herbal medicines (HMs). However, discovery of labeled constituents that can account for the whole efficacy of original HMs is a challenging issue. Taking tanshinones extract (TE) as an example, a strategy to establish reasonable quality control method using effective compounds combination (ECC) as labeled constituents was proposed. The strategy consists of three core steps, including chemical profiling of TE, discovery and in vivo process research of ECC, and quality standardization based on ECC. Using this strategy, a combination of four tanshinones (tanshinones IIA, cryptotanshinone, tanshinone I and dihydrotanshinone I), which was as effective as TE in cell models and in a rat model of myocardial ischemia-reperfusion, was identified as ECC in TE. Furthermore, quality standardization of TE was established based on proposed ECC using single standard for determination of multiple components (SSDMC) method. In conclusion, the presently developed ECC-based approach not only offers new insight into the understanding of the holistic effects of HMs, but also provides efficacy-associated labeled constituents for quality control of botanicals.

Asunto(s)

Abietanos/química , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Plantas Medicinales/química , Abietanos/farmacología , Animales , Modelos Animales de Enfermedad , Medicina de Hierbas/métodos , Masculino , Isquemia Miocárdica/tratamiento farmacológico , Miocitos Cardíacos/efectos de los fármacos , Control de Calidad , Ratas , Ratas Sprague-Dawley , Estándares de Referencia

RESUMEN

Citri Reticulatae Pericarpium, mainly including the pericarp of Citrus reticulata Blanco and the pericarp of Citrus reticulata 'Chachi', has been consumed daily as food and dietary supplement for centuries. In this study, GC-MS based metabolomics was employed to compare comprehensively the volatile constituents in Citrus reticulata Blanco and Citrus reticulata 'Chachi'. Principal component analysis and orthogonal partial least squares discrimination analysis indicated that samples could be distinguished effectively from one another. Fifteen metabolites were finally identified for use as chemical markers in discrimination of Citri Reticulatae Pericarpium samples. The antimicrobial activity against Gram-negative and Gram-positive bacteria of the volatile oil from Citrus reticulata Blanco and Citrus reticulata 'Chachi' was investigated preliminarily.

Asunto(s)

Citrus/química , Metabolómica , Aceites Volátiles/análisis , Cromatografía de Gases y Espectrometría de Masas

RESUMEN

Xingxiong injection (XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacological studies, chemical analysis and quality control studies on this formula are relatively limited. In the present study, a high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in XXI. According to the fragmentation rules and previous reports, thirty ginkgo flavonoids, four ginkgo terpene lactones, and one alkaloid were identified. A high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QQQ MS) method was then applied to quantify ten major constituents in XXI. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. The total contents of ginkgo flavonoids were about 22.05-25.51 µg·mL(-1) and the ginkgo terpene lactones amounts were about 4.41-8.70 µg·mL(-1) in six batches of XXI samples, respectively. Furthermore, cosine ratio algorithm and distance measurements were employed to evaluate the similarity of XXI samples, and the results demonstrated a high-quality consistency. This work could provide comprehensive information on the quality control of Xingxiong injection, which be helpful in the establishment of a rational quality control standard.

Asunto(s)

Alcaloides/análisis , Medicamentos Herbarios Chinos/química , Flavonoides/análisis , Ginkgo biloba/química , Lactonas/análisis , Hojas de la Planta/química , Terpenos/análisis , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos

RESUMEN

A high expression ß1AR/cell membrane chromatography (ß1AR-CMC) and offline UPLC/MS method has been developed for screening active ingredients from Coptis chinensis. In this study, the fractions retained by CMC column were separated and identified by UPLC/MS system. Using metoprolol as a positive control drug, coptisine from C. chinensis was identified as the active component which could inhibit ß1AR. Compared with the control group: coptisine could attenuate the infarct size and release malondialdehyde (MDA) while increasing superoxide dismutase (SOD) activity, suggesting a role in reducing myocardial injury. In vitro, coptisine could decrease apoptosis, showing their protective effects upon cardiomyocytes. This ß1AR-CMC-offline-UPLC/MS method can be applied for screening active components acting on ß1AR from traditional Chinese medicines.

Asunto(s)

Antagonistas de Receptores Adrenérgicos beta 1/farmacología , Berberina/análogos & derivados , Membrana Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Coptis , Espectrometría de Masas , Extractos Vegetales/farmacología , Receptores Adrenérgicos beta 1/efectos de los fármacos , Antagonistas de Receptores Adrenérgicos beta 1/aislamiento & purificación , Antagonistas de Receptores Adrenérgicos beta 1/metabolismo , Animales , Apoptosis/efectos de los fármacos , Berberina/aislamiento & purificación , Berberina/metabolismo , Berberina/farmacología , Unión Competitiva , Células CHO , Membrana Celular/metabolismo , Coptis/química , Cricetulus , Modelos Animales de Enfermedad , Masculino , Malondialdehído/metabolismo , Metoprolol/metabolismo , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Infarto del Miocardio/prevención & control , Miocardio/metabolismo , Miocardio/patología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Fitoterapia , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/metabolismo , Plantas Medicinales , Ratas Wistar , Receptores Adrenérgicos beta 1/genética , Receptores Adrenérgicos beta 1/metabolismo , Superóxido Dismutasa/metabolismo , Transfección

RESUMEN

Miltirone is one of the bioactive diterpene quinones isolated from Salvia miltiorrhiza Bunge. This compound has been found to possess significant anticancer, antibacterial, antioxidant, and anti-inflammatory activities. However, the metabolic fate of miltirone remains unknown. In order to explore whether miltirone is extensively metabolized, we investigated the metabolites of miltirone in plasma, bile, urine, and feces samples following oral and intravenous administration to the rats. By using high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF-MS) coupled with mass detect filter (MDF) method, a total of 15 metabolites were identified from the biosamples. Both phase I and phase II metabolites were observed in the metabolic profile and the metabolic pathways involved in reduction, oxidation, monohydroxylation, dihydroxylation, glucuronidation and sulfation. The results indicated that hepatocyte metabolism was the major route of clearance for the parent compound. The present study provided valuable information for better understanding of the efficacy and safety of miltirone.

Asunto(s)

Metaboloma/fisiología , Fenantrenos/metabolismo , Animales , Bilis/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Heces/química , Masculino , Espectrometría de Masas/métodos , Fenantrenos/sangre , Fenantrenos/orina , Ratas , Ratas Sprague-Dawley

RESUMEN

Citri Fructus (CF), the mature fruit of Citrus wilsonii Tanaka (CWT) or Citrus medica L. (CML), is an important citrus by-product with health promoting and nutritive properties. The present study compares the chemical and biological differences of CWT and CML. Thin layer chromatography and high performance liquid chromatography, coupled with quadrupole time-of-flight tandem mass spectrometry techniques, were employed to compare the chemical profiles of CWT and CML. A total of 25 compounds were identified and the results indicated that there were significant differences in chemical composition between the two CF species. The quantitative results obtained by HPLC coupled with diode array detector method demonstrated that naringin was present in the highest amounts in CWT, whilst nomilin was the most dominant constituent in CML. It was also found that CWT had significantly higher free radical-scavenging activity than CML.

Asunto(s)

Antioxidantes/análisis , Citrus/química , Frutas/química , Extractos Vegetales/análisis , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos

RESUMEN

A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of miltirone concentration in rat plasma. The cytotoxic activity of miltirone was firstly evaluated by the MTT assay and compared with other tanshinones. Quantification was carried out on an Agilent triple quadrupole LC-MS system using multiple reaction monitoring (MRM) mode in positive mode. After simple protein precipitation with acetonitrile, the chromatographic separation of miltirone was achieved by using a Waters Symmetry C18 analytical column (2.1mm×100mm, 3.5µm) with a mobile phase of acetonitrile (A)-water (B) (75:25, v/v) containing 0.5% formic acid. The monitored transitions were set at m/z 283.1→223.1 and m/z 361.0→232.9 for miltirone and IS, respectively. The calibration curve was linear over the concentration range of 0.5-200ng/mL with lower limit of quantification of 0.5ng/mL. The intra- and inter-day accuracy and precision of miltirone were both within acceptable limits. The developed method was successfully applied to a pharmacokinetic study of following oral administration of 20, 40, 60mg/kg and an intravenous administration of 0.5mg/kg to rats. The results indicated that miltirone had linear pharmacokinetic properties within the tested dosage range and was poorly absorbed with an absolute bioavailability of approximately 3.4%.

RESUMEN

Citrus grandis 'Tomentosa' (CGT) is particularly cultivated in China and widely used in health foods. In this study, the chemical profiles of different parts of CGT were comprehensively compared by rapid resolution liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry method. A total of 22 compounds were identified and two C-glucosyl flavones were found for the first time in CGT. Four main constituents (rhiofolin, naringin, meranzin hydrate, and isoimperatorin) in different parts of CGT were simultaneously determined. Overall, the contents of the four main compounds decreased with the ripening process. In parallel, the antioxidant activities of their extracts were also evaluated by three assays (2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) diammonium salt, 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power), and the results indicated a similar tendency: small fruit > flower ∼ medium fruit > large fruit > leaf ∼ branch. The results obtained in the present work may provide useful information for future utilization of CGT.

Asunto(s)

Antioxidantes/química , Citrus/química , Extractos Vegetales/química , Flores/química , Alimentos Orgánicos/análisis , Frutas/química , Hojas de la Planta/química , Tallos de la Planta/química , Espectrometría de Masa por Ionización de Electrospray

RESUMEN

Although traditional Chinese medicines (TCMs) play important role in drug discovery and human health, the actual value of TCMs has not been fully recognized worldwide due to its complex components and uncontrollable quality. For the modernization and globalization of TCMs, it is important to establish selective, sensitive and feasible analytical methods for determination and quantification of bioactive components of TCMs in body fluids primarily due to the low concentration, the complex nature of the biological matrices, and multi-components and their metabolites present in biological fluids. The present review summarizes the current extraction techniques, chromatographic separation and spectroscopic (especially mass spectrometric) analysis methods and new trends on the analysis of bioactive components and metabolites of TCMs in biological fluids. In addition, the importance of establishment of pharmacokinetics and bioavailability profiles and simultaneous determination of multi-active components in TCMs is discussed to provide proper examples of analytical methods for pharmacological and clinical studies of TCMs.

Asunto(s)

Técnicas de Química Analítica/métodos , Medicamentos Herbarios Chinos/análisis , Medicina Tradicional China/normas , Animales , Disponibilidad Biológica , Líquidos Corporales/química , Descubrimiento de Drogas , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacocinética , Humanos , Control de Calidad

RESUMEN

We report a case of continuous ambulatory peritoneal dialysis (CAPD)-related peritonitis. Burkholderia gladioli, as a plant pathogen, were cultured from peritoneal effluent. Peritonitis healed after intraperitoneal cefazolin and gentamicin. This case indicated that CAPD effluent may be contaminated by B. gladioli, causing CAPD-related peritonitis. On the other hand, it can be successfully treated with intraperitoneal antimicrobials without removal of the Tenckhoff catheter.

Asunto(s)

Infecciones por Burkholderia/patología , Burkholderia gladioli/aislamiento & purificación , Peritonitis/microbiología , Anciano , Humanos , Masculino , Diálisis Peritoneal Ambulatoria Continua/efectos adversos