ABSTRACT
A fast and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of prodrug of paclitaxel (Pro-PTX) and paclitaxel (PTX) in rat plasma was developed. The plasma samples were subjected to protein precipitation with acetonitrile (0.1% formic acid), and then separated by LC with an Ultimate AQ-C18 column (50 mm × 3.0 mm, 3 μm) and acetonitrile-1 mmol·L-1 ammonium formate (containing 0.1% formic acid) as the mobile phase. Multiple reaction monitoring (MRM) scanning mode was used to detect the ion responses m/z 983.4→415.2 (Pro-PTX), m/z 854.4→286.1 (PTX) and m/z 808.3→527.2 (Docetaxel, internal standard) by using a triple quadrupole tandem mass spectrometer with electrospray ionization source and positive ion mode. The method validation results show that the linear ranges of Pro-PTX and PTX in plasma were 2.00-500 ng·mL-1 (r > 0.99) and 4.00-1 000 ng·mL-1 (r > 0.99), the lower limit of quantification was 2.00 ng·mL-1 and 4.00 ng·mL-1, respectively; the quality control samples with low, medium and high concentrations of Pro-PTX and PTX were within the batch, the relative standard deviation (RSD) between batches were all less than 9%; the relative deviation (RE) was within the range of ± 9%; In the stability test, both Pro-PTX and PTX in plasma were stable under various storage conditions. The method was sensitive, specific, and reproducible, and was suitable for the pharmacokinetic study of Pro-PTX in rats. Animal experiments were approved by the Ethics Committee of Laboratory Animal Management and Animal Welfare, Institute of Materia Medica, Chinese Academy of Medical Sciences (No.: 00003552).
ABSTRACT
Multicellular tumor spheroids (MCTS) can simulate the structure and metabolic characteristics of tumors in vivo, which is of great significance to study the metabolic phenotype of tumor cells and the mechanism of drug intervention. In this study, esophageal cancer MCTS were constructed, and MCTS frozen sections were prepared after treated with different formulations of paclitaxel (PTX) including common PTX injection, PTX liposome and albumin bound PTX. MCTS mass spectrometry imaging analysis method was established by using air flow assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). The visualization of the permeation and enrichment process of PTX in MCTs after PTX treatment was realized, and the spatially resolved metabolomics of PTX injection group was studied. The results showed that the permeation and enrichment behavior of PTX in MCTs model were related to the formulations. The changes of endogenous metabolites in MCTs of esophageal cancer after treated with PTX injection had temporal and spatial characteristics. The metabolic changes of MCTS during the initial 0-4 hours were dominated by the down-regulation of middle-high polarity metabolites and some lipids in the central region of MCTS, while the metabolic changes of MCTS during 8-72 hours were mainly up-regulated by lipid metabolites in the peripheral region of MCTS. The combination of in vivo tumor-associated MCTs model with label free, highly sensitive and high coverage mass spectrometry imaging technology provided a new method and strategy for the study of pharmacometabolomics.
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In order to achieve rapid proliferation and adapt to the complex microenvironment, tumor cells have dominant characteristics such as unique metabolic patterns and the ability to escape from immunoregulation. Tumor cells reprogram multiple metabolic pathways to promote immune escape, which impacts tumor diagnosis, treatment and prognosis. Based on the effect of metabolic changes on tumor immune escape and its molecular mechanism, metabolic regulation provides new approaches to enhance immunotherapy. We review recent advances in tumor immuno-escape and immunotherapy based on metabolic regulation. Cutting-edge analytical techniques and methods for tumor metabolism research such as metabolomics, mass spectrometry imaging-based spatially-resolved metabolomics and metabolic flow analysis are also discussed.
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As a promising new molecular imaging technique,mass spectrometry imaging(MSI) has attracted more and more attention in the field of biomedicine. A method of air flow assisted ionization-ultra high resolution mass spectrometry-based mass spectrometric imaging (AFAI-MSI) was developed to profile endogenous metabolites in rat kidney tissue in this study. Rat kidneys were collected and cut into frozen tissue sections,and then were analyzed on an AFAI-MSI system in positive ion mode using acetonitrile-isopmpyl alcohol-water (4:4:2,V/V,5 μL/min) as spray solvent,N2as spray gas(0.6 MPa) and air as assisting gas (45 L/min). The mass range and resolution were set to be 70-1000 Da and 70000, respectively. As a result,a total of 38 metabolites, including organic amines, sugars, vitamins, peptides, neurotransmitters, organic acids,phospholipids,sphingolipids,glyceride,and cholesterol esters, were identified and imaged to characterize their tissue-specific distribution in kidney tissues, and some metabolites, such as choline, acetylcoline,betaine,phoshocholine,and glycerophosphocholine were found to have distinct distribution along the cortex-medulla axis,which may be involved in the formation of osmotic pressure gradient in the kidney. The proposed ultra high resolution mass spectrometry based AFAI-MSI method could work without sample pretreatment, showed high sensitivity and wide metabolite coverage, and was expected to provide a new analytical approach for the research of in situ characterization and metabolic regulation mechanism of endogenous metabolites in kidney.
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Due to the diversity and complexity, the change of chemical components in medicinal plant according to the time, cultivated varieties or ecological condition is difficult to recognize using traditional phytochemistry method. In order to analyze the pharmacodynamics material basis in Uighur medicinal plant Artemisia rupestris L. in an effective and comprehensive way, a plant metabolomics approach was established based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). This study firstly focused on the effect of extraction solvents,redissolve solvents and ultrasonic time on the untargeted metabolomics, then the optimal preparation condition was selected according to metabolites coverage. After methodology validation, the approach was applied to acquire metabolic information in root, stem, branchlet, leaf and flower of Artemisia rupestris L. The results showed that the metabolome in flower was obviously different with the other organs. Coupling with multivariate statistical analysis, a batch of differential metabolites were picked out, in which 61 flavonoids, 97 rupestonic acid derivatives, 7 chlorogenic acids and 15 other compounds were primarily recognized according to the characteristic fragmentation rules of specific structure type and database retrieval. Additionally,the distribution characteristics of the above 180 differential metabolites was illustrated by cluster heat map. In conclusion,this study provided important information about the rational utilization of effective parts from Artemisia rupestris L.,and offered a novel strategy for quality control,variety improvement and reasonable development of medicinal plants.
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The study aims to develop a rapid, specific and sensitive method for quantitative analysis of trace impurities in levofloxacin formulation using LC-MS/MS. The quality of the different formulations from 19 plants was evaluated in the contents of the impurities. The results indicated that there were 5 impurities in the samples, and the content was different in the products with same formulation by different plants. The products of 3 plants were in good quality with impurities level under 0.01%. Levofloxacin N4'-methyl quaternary impurity was first reported as the formulation impurity. The impurities were tightly correlated to the reservation of drug, process control of formulation and storage during transportation. The results suggest that our method is sensitive and specific to detect the trace impurities in formulation, and can be used to monitor the quality of commercial drug product.
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An HPLC-DAD-MS/MS method was developed for rapid analysis and identification of degradation products of buagafuran. Buagafuran and degradation products were separated on a Zorbax C8 column (5 microm, 4.6 mm x 150 mm) using acetonitrile-water (78 : 22) as mobile phase. The elutes were detected with diode array detector and tandem mass spectrometer via electrospray ionization source in positive ion mode. According to analysis of the retention time, UV spectra and MS, MS/MS data, combined with the possible degradation reaction of buagafuran, the structures of main degradation products were inferred. The results showed that six main degradation products were oxidation or peroxidation productions of buagafuran. Degradation product A was a double bond epoxidation product of buagafuran, degradation products B, C, D and E were the further oxidation products of degradation product A, degradation product F was a peroxidation product of buagafuran. The results indicated that the established method was effective in the rapid identification of the degradation products of buagafuran.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Sesquiterpenes , Chemistry , Spectrometry, Mass, Electrospray Ionization , Methods , Tandem Mass Spectrometry , MethodsABSTRACT
To screen the harmful substance 5-hydroxymethyl furfural content in commercially available traditional Chinese medicine injection which are commonly used, and to preliminarily evaluate the quality of these injections, 5-hydroxymethyl furfural was taken as an index. The contents of 5-hydroxymethyl furfural in 56 samples which consist of 23 kinds of traditional Chinese medicine injections and glucose injection were determined using LC-MS/MS, and 5-hydroxymethyl furfural was detected in 52 of these samples. The minimal content was 0.0038 microg x L(-1) and the maximum content was 1420 microg x mL(-1). The contents of 5-hydroxymethyl furfural were significantly different in traditional Chinese medicine injection which came from different kinds, manufacturers or batches. The results showed the quality difference of commercially available traditional Chinese medicine injection is significant taking 5-hydroxymethyl furfural content as assessment index. More attention should be paid to the safety of 5-hydroxymethyl furfural in traditional Chinese medicine injection, and unified limitation standard should be set to improve medication safety of traditional Chinese medicine injection.
Subject(s)
Chromatography, Liquid , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Eleutherococcus , Chemistry , Furaldehyde , Glucose , Chemistry , Injections , Medicine, Chinese Traditional , Plant Extracts , Chemistry , Plants, Medicinal , Chemistry , Tandem Mass SpectrometryABSTRACT
To study the condensation mechanism of sodium new houttuyfonate, and determinate the chemical structure of condensation products, dimer was prepared, and LC-DAD-MS/MS multiple techniques were employed to investigate the ultraviolet absorption feature and mass spectrum of transformation solution of dimer, and the transformation kinetics and half-life were studied by ultraviolet spectrophotometry. The pure substance of stable condensation product was obtained by extracting with organic solvent and purifying with column chromatography, the chemical structure of this substance was identified by assaying of IR, HR-ESI-MS and NMR, and the data of LC-MS/MS were compared with that of transformation products of dimer. The results indicated that the dimer is unstable, it will be rapidly dissociated in aqueous solution to form free new houttuyfonate and then cycloaddition reaction will occur and followed by an in situ dehydration to generate 1, 3, 5-tri (dodecanoyl) benzene (trimer) with a six-ring which is stable in aqueous solution. The transformation process may fit second-order kinetics, and the half-times were found to be 3.17 hours at 25 degrees C (298 K) and 6.39 min at 100 degrees C (373 K), separately. It suggests that dimer is an intermediate in condensation reaction, and the end condensation product of sodium new houttuyfonate injection may exist as trimer.
Subject(s)
Alkanes , Chemistry , Pharmacology , Chromatography, Liquid , Molecular Structure , Pharmaceutical Preparations , Sulfites , Chemistry , Pharmacology , Tandem Mass SpectrometryABSTRACT
In this review, we provide information on hydroxyl radical generation, trapping and detection methods, including electron spin resonance (ESR), electrochemistry detection (ECD), fluorescence detection, UV detection, chemoluminescence and mass spectrometry (MS). In addition, the advantages and disadvantages of the above methods were discussed.