Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and pathological conditions.Her...Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and pathological conditions.Herein,airflow-assisted desorption electrospray ionization-MSI(AFADESI-MSI)was coupled with 3D HepG2 spheroids to assess the metabolism and hepatotoxicity of amiodarone(AMI).High-coverage imaging of>1100 endogenous metabolites in hepatocyte spheroids was achieved using AFADESI-MSI.Following AMI treatment at different times,15 metabolites of AMI involved in Ndesethylation,hydroxylation,deiodination,and desaturation metabolic reactions were identified,and according to their spatiotemporal dynamics features,the metabolic pathways of AMI were proposed.Subsequently,the temporal and spatial changes in metabolic disturbance within spheroids caused by drug exposure were obtained via metabolomic analysis.The main dysregulated metabolic pathways included arachidonic acid and glycerophospholipid metabolism,providing considerable evidence for the mechanism of AMI hepatotoxicity.In addition,a biomarker group of eight fatty acids was selected that provided improved indication of cell viability and could characterize the hepatotoxicity of AMI.The combination of AFADESI-MSI and HepG2 spheroids can simultaneously obtain spatiotemporal information for drugs,drug metabolites,and endogenous metabolites after AMI treatment,providing an effective tool for in vitro drug hepatotoxicity evaluation.展开更多
Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity ...Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.展开更多
Eight metabolites of brodimoprim (BDP) in rat urine were detected by NMR and ESIMS/MS. They were demethyl-BDP glucuronide, demethyl-BDP sulfurate, demethyl-BDP glucuronide sulfurate, alpha -hydroxyl-BDP, alpha -hydrox...Eight metabolites of brodimoprim (BDP) in rat urine were detected by NMR and ESIMS/MS. They were demethyl-BDP glucuronide, demethyl-BDP sulfurate, demethyl-BDP glucuronide sulfurate, alpha -hydroxyl-BDP, alpha -hydroxyl-BDP glucuronide, BDP sulfurate, N-oxide-BDP sulfurate, and alpha -hydroxyl-N-oxide-BDP sulfurate. All the sulfurates are reported for the first time.展开更多
A novel ambient glow discharge ion source with improved line-cylinder electrodes is put forward and designed in this paper. The diameters of inner and outer electrodes are 0.16 and 4 mm respectively. With a special as...A novel ambient glow discharge ion source with improved line-cylinder electrodes is put forward and designed in this paper. The diameters of inner and outer electrodes are 0.16 and 4 mm respectively. With a special assembly method, a perfect coaxiality of the two electrodes is obtained. From the gas discharge experiment, it can be seen that the discharge can stably work in normal glow discharge mode. The operating currents of the ion source are in an order of milliamperes and can generate a much larger number and wider variety of reagent ions. The MS experiment shows that the ion source has higher detection sensitivity,展开更多
The mechanism of ionization and fragmentation for terphenyl (diphenylbenzene) with three structural ring isomers (ortho-, meta- and para-), and stilbene (1,2-diphenylethylene) with two geometrical isomers (trans- and ...The mechanism of ionization and fragmentation for terphenyl (diphenylbenzene) with three structural ring isomers (ortho-, meta- and para-), and stilbene (1,2-diphenylethylene) with two geometrical isomers (trans- and cis-) by EI mass spectrometry and ionization efficiency curves are investigation.展开更多
Deconvolution of potential drug targets of the central nervous system(CNS)is particularly challenging because of the complicated structure and function of the brain.Here,a spatiotemporally resolved metabolomics and is...Deconvolution of potential drug targets of the central nervous system(CNS)is particularly challenging because of the complicated structure and function of the brain.Here,a spatiotemporally resolved metabolomics and isotope tracing strategy was proposed and demonstrated to be powerful for deconvoluting and localizing potential targets of CNS drugs by using ambient mass spectrometry imaging.This strategy can map various substances including exogenous drugs,isotopically labeled metabolites,and various types of endogenous metabolites in the brain tissue sections to illustrate their microregional distribution pattern in the brain and locate drug action-related metabolic nodes and pathways.The strategy revealed that the sedative-hypnotic drug candidate YZG-331 was prominently distributed in the pineal gland and entered the thalamus and hypothalamus in relatively small amounts,and can increase glutamate decarboxylase activity to elevateγ-aminobutyric acid(GABA)levels in the hypothalamus,agonize organic cation transporter 3 to release extracellular histamine into peripheral circulation.These findings emphasize the promising capability of spatiotemporally resolved metabolomics and isotope tracing to help elucidate the multiple targets and the mechanisms of action of CNS drugs.展开更多
Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity.Miao-Fu-Zhi-Tong(MFZT)granules,an ethnic Yi herbal prescription,comprises 10 herbs...Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity.Miao-Fu-Zhi-Tong(MFZT)granules,an ethnic Yi herbal prescription,comprises 10 herbs and has been clinically applied for gouty arthritis(GA)therapy.Herein,a series of chemical profiling strategies including in-house library matching,molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)were developed for qualitative analysis of MFZT granules.A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes.Moreover,network pharmacology analysis indicated that MFZT’s anti-gout mechanism was mostly associated with the nuclear factor kappa-B(NF-κB)signaling,nucleotide oligomerization domain(NOD)-like signaling and rheumatoid arthritis pathways,along with the synergistic effect of 84 potential active compounds.In addition,a quantitative analytical method was developed to simultaneously determine the 29 potential effective components.Among them,berberine,pellodendrine,3-feruloylquinic acid,neoastilbin,isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control.These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.展开更多
On-tissue chemical derivatization(OTCD)effectively enhances ionization efficiency of low abundant and poorly ionized functional molecules to improve detection sensitivity and coverage of mass spectrometry imaging(MSI)...On-tissue chemical derivatization(OTCD)effectively enhances ionization efficiency of low abundant and poorly ionized functional molecules to improve detection sensitivity and coverage of mass spectrometry imaging(MSI).Combination OTCD and MSI provides a novel strategy for visualizing previously undisclosed metabolic heterogeneity in tumor.Herein,we present a method to visualize heterogeneous metabolism of oxylipins within tumor by coupling OTCD with airflow-assisted desorption electrospray ionization(AFADESI)-MSI.Taking Girard’s P as a derivatization reagent,easily ionized hydrazide and quaternary amine groups were introduced into the structure of carbonyl metabolites via condensation reaction.Oxylipins,including 127 fatty aldehydes(FALs)and 71 oxo fatty acids(FAs),were detected and imaged in esophageal cancer xenograft with AFADESI-MSI after OTCD.Then t-distributed stochastic neighbor embedding and random forest were exploited to precisely locate the distribution of oxylipins in heterogeneous tumor tissue.With this method,we surprisingly found almost all FALs and oxo FAs significantly accumulated in the core region of tumor,and exhibited a gradual increase trend in tumor over time.These results reveal spatiotemporal heterogeneity of oxylipins in tumor progression,highlighting the value of OTCD combined with MSI to gain deeper insights into understanding tumor metabolism.展开更多
Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy(DN)is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies.In the...Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy(DN)is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies.In the present study,a spatial-resolved metabolomics approach based on air flowassisted desorption electrospray ionization(AFADESI)and matrix-assisted laser desorption ionization(MALDI)integrated mass spectrometry imaging(MSI)was proposed to investigate tissue-specific metabolic alterations in the kidneys of high-fat diet-fed and streptozotocin(STZ)-treated DN rats and the therapeutic effect of astragalosideⅣ,a potential anti-diabetic drug,against DN.As a result,a wide range of functional metabolites including sugars,amino acids,nucleotides and their derivatives,fatty acids,phospholipids,sphingolipids,glycerides,carnitine and its derivatives,vitamins,peptides,and metal ions associated with DN were identified and their unique distribution patterns in the rat kidney were visualized with high chemical specificity and high spatial resolution.These region-specific metabolic disturbances were ameliorated by repeated oral administration of astragaloside Ⅳ(100 mg/kg)for 12 weeks.This study provided more comprehensive and detailed information about the tissue-specific metabolic reprogramming and molecular pathological signature in the kidney of diabetic rats.These findings highlighted the promising potential of AFADESI and MALDI integrated MSI based metabolomics approach for application in metabolic kidney diseases.展开更多
Understanding of the nephrotoxicity induced by drug candidates is vital to drug discovery and development.Herein,an in situ metabolomics method based on air flow-assisted desorption electrospray ionization mass spectr...Understanding of the nephrotoxicity induced by drug candidates is vital to drug discovery and development.Herein,an in situ metabolomics method based on air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI)was established for direct analysis of metabolites in renal tissue sections.This method was subsequently applied to investigate spatially resolved metabolic profile changes in rat kidney after the administration of aristolochic acid I,a known nephrotoxic drug,aimed to discover metabolites associated with nephrotoxicity.As a result,38 metabolites related to the arginine-creatinine metabolic pathway,the urea cycle,the serine synthesis pathway,metabolism of lipids,choline,histamine,lysine,and adenosine triphosphate were significantly changed in the group treated with aristolochic acid I.These metabolites exhibited a unique distribution in rat kidney and a good spatial match with histopathological renal lesions.This study provides new insights into the mechanisms underlying aristolochic acids nephrotoxicity and demonstrates that AFADESI-MSI-based in situ metabolomics is a promising technique for investigation of the molecular mechanism of drug toxicity.展开更多
With the rapid development and wide application of traditional Chinese medicine injection(TCMI), a number of adverse events of some TCMIs have incessantly been reported and have drawn broad attention in recent years. ...With the rapid development and wide application of traditional Chinese medicine injection(TCMI), a number of adverse events of some TCMIs have incessantly been reported and have drawn broad attention in recent years. Establishing effective and practical analytical methods for safety evaluation and quality control of TCMI can help to improve the safety of TCMIs in clinical applications. In this study, a sensitive and rapid high-performance liquid chromatography–tandem mass spectrometry(HPLC–MS/MS)method has been developed and validated for the quantitative determination of potentially harmful substance5,5′-oxydimethylenebis(2-furfural, OMBF) in TCMI samples. Chromatographic separation was performed on a C18 reversed-phase column(150 mm × 2.1 mm, 5 μm) by gradient elution, using methanol–water containing 0.1% formic acid as mobile phase at the flow rate of 0.3 m L/min. MS/MS detection was performed on a triple quadrupole mass spectrometer with positive electrospray ionization in the multiple reaction-monitoring mode. The method was sensitive with a limit of quantification of 0.3 ng/m L and linear over the range of 0.3–30 ng/m L(r = 0.9998). Intra-and inter-day precision for analyte was o9.52% RSD withrecoveries in the range 88.0–109.67% at three concentration levels. The validated method was successfully applied to quantitatively determine the compound OMBF in TCMIs and glucose injections. Our study indicates that this method is simple, sensitive, practicable and reliable, and could be applied for safety evaluation and quality control of TCMIs and glucose injections.展开更多
Mass spectrometry imaging (MSI) technology can simultaneously obtain the spatial distribution of thousands of chemical compounds and has unique advantages compared to other techniques that allow mapping the surface of...Mass spectrometry imaging (MSI) technology can simultaneously obtain the spatial distribution of thousands of chemical compounds and has unique advantages compared to other techniques that allow mapping the surface of bio-tissue. Here, we combined an air flow-assisted desorption electrospray ionization (AFADESI) MSI device with a high-resolution mass spectrometer to optimize the system parameters and achieve more accurate spatial distribution characteristics for compounds of interest while investigating bio-tissue sections. The platform set-up, required instrumentation, sample pretreatment, parameter optimization and bio-tissue characterization are described and discussed.Finally, the parameter conditions that can provide optimal ionic intensity and enhanced resolution were confirmed. The reasonable resolution and sensitivity improvements of AFADESI-MSI have been achieved through tandem a high-resolution mass spectrometer system, therefore, it would be a promising technique for the bio-tissue imaging analysis.展开更多
A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry(UPLC–MS/MS) method was developed and fully validated for determination of arformoterol in rat plasma, lung and trachea tissues....A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry(UPLC–MS/MS) method was developed and fully validated for determination of arformoterol in rat plasma, lung and trachea tissues. The chromatographic separation was performed on an Agilent XDB C8 column with gradient elution by using acetonitrile and 0.1% formic acid water. The method presented high sensitivity(LLOQ of 1.83 pg/mL for plasma and 3.90 pg/mL for lung and trachea tissue homogenates) and great linearity over the range of 1.83–458 pg/mL for plasma, 13.9–1560 pg/mL for lung and trachea tissue homogenates. No carry over effect and matrix effect were observed. The intra-and inter-day precision/accuracy were within ±15% at three quality control concentration levels. This robust method was successfully applied to the pharmacokinetic, lung and trachea tissue distribution study after inhalation administration of arformoterol tartrate inhalation solution(50 m g/kg). The in vivo information indicate that arformoterol can be rapidly absorbed into blood through respiratory systems, lung and trachea tissue maintain a certain amount of arformoterol in 1 h after dosing.展开更多
We have studied the fragmentation behavior of positive and negative ions of taxoi and 6/8/6 type taxoids, the influence of different substituents on fragmentation, and the correlativity between fragmentation patterns ...We have studied the fragmentation behavior of positive and negative ions of taxoi and 6/8/6 type taxoids, the influence of different substituents on fragmentation, and the correlativity between fragmentation patterns and structure by MS/MS technique with different ionization methods such as FAB-MS, ESI-MS, ete. We have also investigated in detail the fragmentation of various molecular-related ions, such as [M+H]+, [M+Na]+ and [M-H] ions, and the formation pathways of characteristic fragment ions. It has been found that there exists some competing reaction between the loss of C-13 side chain and decomposition by loss of acetic acid. In addition, by comparing CID spectra obtained with low- and high-energy collision, it is seen that CID-MS/MS with low-energy collision is more suitable for the study of the structural analysis of small molecules and drug metabolites. The experimental results demonstrate that MS/MS spectra can reflect more effectively the slight difference of structure between the related展开更多
The imaging mass spectrometry(IMS) technology has experienced a rapid development in recent years.A new IMS technology which is based on air flow assisted ionization(AFAI) was reported.It allows for the convenient...The imaging mass spectrometry(IMS) technology has experienced a rapid development in recent years.A new IMS technology which is based on air flow assisted ionization(AFAI) was reported.It allows for the convenient pretreatment of the samples and can image a large area of sample in a single measurement with high sensitivity.The AFAI in DESI mode was used as the ion source in this paper.The new IMS method is named AFADESI-IMS.The adoption of assisted air flow makes the sample pretreatment easy and convenient.An optimization of the distance between the ion transport tube and MS orifice increases the sensitivity of the system.For data processing,a program based on MATLAB with the function of numerical analysis was developed.A theoretical imaging resolution of a few hundred microns can be achieved.The composite AFAI-IMS images of different target analytes were imaged with high sensitivity.A typical AFAI-IMS image of the whole-body section of a rat was obtained in a single analytical measurement.The ability to image a large area for relevant samples in a single measurement with high sensitivity and repeatability is a significant advantage.The method has enormous potentials in the MS imaging of large and complicated samples.展开更多
A new multivariate statistical strategy for analyzing large datasets that are produced by imaging mass spectrometry(IMS) techniques is reported.The strategy divides the whole datacube of the sample into several subs...A new multivariate statistical strategy for analyzing large datasets that are produced by imaging mass spectrometry(IMS) techniques is reported.The strategy divides the whole datacube of the sample into several subsets and analyses them one by one to obtain the results.Instead of analyzing the whole datacube at one time,the strategy makes the analysis easier and decreases the computation time greatly.In this report,the IMS data are produced by the air flow-assisted ionization IMS(AFAI-IMS).The strategy can be used in combination with most multivariate statistical analysis methods.In this paper,the strategy was combined with the principal component analysis(PCA) and partial least square analysis(PLS).It was proven to be effective by analyzing the handwriting sample.By using the strategy,the m/z corresponding to the specific lipids in rat brain tissue were distinguished successfully.Moreover the analysis time grew linearly instead of exponentially as the size of sample increased.The strategy developed in this study has enormous potential for searching for the mjz of potential biomarkers quickly and effectively.展开更多
A ’H NMR signal shifted drastically to down field (δ - 10.0) at the bay area and in dose proximity to C = O group for H-1 was observed through complete assignments of 1H NMR spectra for pyridino- and benzobenzanthro...A ’H NMR signal shifted drastically to down field (δ - 10.0) at the bay area and in dose proximity to C = O group for H-1 was observed through complete assignments of 1H NMR spectra for pyridino- and benzobenzanthrones. It is concluded that this phenomenon is due not only to the anisotropy effect of C=O plus aromatic ring current effect, but also to the electrostatic attraction of C-H (δ+)…O(δ-)=C interaction. The evidence for the sp2C-H…O = C intramolecular interaction similar to hydrogen bond has also been given by EI-MS and MS/MS (CID) spectroscopy and IR spectroscopy, as well as MNDO-PM3 calculations. This new kind of interaction might be called ’quasi-hydrogen bond’.展开更多
基金funded by the National Natural Science Foundation of China(Grant No.:21874156)the Chinese Academy of Medical Science(CAMS)Innovation Fund for Medical Sciences(Grant No.:2021-1-I2M-028).
文摘Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and pathological conditions.Herein,airflow-assisted desorption electrospray ionization-MSI(AFADESI-MSI)was coupled with 3D HepG2 spheroids to assess the metabolism and hepatotoxicity of amiodarone(AMI).High-coverage imaging of>1100 endogenous metabolites in hepatocyte spheroids was achieved using AFADESI-MSI.Following AMI treatment at different times,15 metabolites of AMI involved in Ndesethylation,hydroxylation,deiodination,and desaturation metabolic reactions were identified,and according to their spatiotemporal dynamics features,the metabolic pathways of AMI were proposed.Subsequently,the temporal and spatial changes in metabolic disturbance within spheroids caused by drug exposure were obtained via metabolomic analysis.The main dysregulated metabolic pathways included arachidonic acid and glycerophospholipid metabolism,providing considerable evidence for the mechanism of AMI hepatotoxicity.In addition,a biomarker group of eight fatty acids was selected that provided improved indication of cell viability and could characterize the hepatotoxicity of AMI.The combination of AFADESI-MSI and HepG2 spheroids can simultaneously obtain spatiotemporal information for drugs,drug metabolites,and endogenous metabolites after AMI treatment,providing an effective tool for in vitro drug hepatotoxicity evaluation.
基金supported by the National Natural Science Foundation of China(Grant Nos.:21927808 and 81803483).
文摘Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.
文摘Eight metabolites of brodimoprim (BDP) in rat urine were detected by NMR and ESIMS/MS. They were demethyl-BDP glucuronide, demethyl-BDP sulfurate, demethyl-BDP glucuronide sulfurate, alpha -hydroxyl-BDP, alpha -hydroxyl-BDP glucuronide, BDP sulfurate, N-oxide-BDP sulfurate, and alpha -hydroxyl-N-oxide-BDP sulfurate. All the sulfurates are reported for the first time.
基金supported by the National Natural Science Foundation of China(No.60706030 and No.20827007)the National High Technology Research and Development Plan of China(No.2007AA04Z337)
文摘A novel ambient glow discharge ion source with improved line-cylinder electrodes is put forward and designed in this paper. The diameters of inner and outer electrodes are 0.16 and 4 mm respectively. With a special assembly method, a perfect coaxiality of the two electrodes is obtained. From the gas discharge experiment, it can be seen that the discharge can stably work in normal glow discharge mode. The operating currents of the ion source are in an order of milliamperes and can generate a much larger number and wider variety of reagent ions. The MS experiment shows that the ion source has higher detection sensitivity,
文摘The mechanism of ionization and fragmentation for terphenyl (diphenylbenzene) with three structural ring isomers (ortho-, meta- and para-), and stilbene (1,2-diphenylethylene) with two geometrical isomers (trans- and cis-) by EI mass spectrometry and ionization efficiency curves are investigation.
基金supported by the National Natural Science Foundation of China(No.21927808 and No.81974500)Chinese Academy of Medical Science(CAMS)Innovation Fund for Medical Sciences(CIFMS,No.2022-I2M-2-002 and 2021-1I2M-028,China)。
文摘Deconvolution of potential drug targets of the central nervous system(CNS)is particularly challenging because of the complicated structure and function of the brain.Here,a spatiotemporally resolved metabolomics and isotope tracing strategy was proposed and demonstrated to be powerful for deconvoluting and localizing potential targets of CNS drugs by using ambient mass spectrometry imaging.This strategy can map various substances including exogenous drugs,isotopically labeled metabolites,and various types of endogenous metabolites in the brain tissue sections to illustrate their microregional distribution pattern in the brain and locate drug action-related metabolic nodes and pathways.The strategy revealed that the sedative-hypnotic drug candidate YZG-331 was prominently distributed in the pineal gland and entered the thalamus and hypothalamus in relatively small amounts,and can increase glutamate decarboxylase activity to elevateγ-aminobutyric acid(GABA)levels in the hypothalamus,agonize organic cation transporter 3 to release extracellular histamine into peripheral circulation.These findings emphasize the promising capability of spatiotemporally resolved metabolomics and isotope tracing to help elucidate the multiple targets and the mechanisms of action of CNS drugs.
基金the National Key Research and Development Program of China(Nos.2017YFC 1704006 and 2017YFC1704005).
文摘Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity.Miao-Fu-Zhi-Tong(MFZT)granules,an ethnic Yi herbal prescription,comprises 10 herbs and has been clinically applied for gouty arthritis(GA)therapy.Herein,a series of chemical profiling strategies including in-house library matching,molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)were developed for qualitative analysis of MFZT granules.A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes.Moreover,network pharmacology analysis indicated that MFZT’s anti-gout mechanism was mostly associated with the nuclear factor kappa-B(NF-κB)signaling,nucleotide oligomerization domain(NOD)-like signaling and rheumatoid arthritis pathways,along with the synergistic effect of 84 potential active compounds.In addition,a quantitative analytical method was developed to simultaneously determine the 29 potential effective components.Among them,berberine,pellodendrine,3-feruloylquinic acid,neoastilbin,isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control.These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.
基金supported by the National Natural Science Foundation of China(No.21927808)the Chinese Academy of Medical Science(CAMS)Innovation Fund for Medical Sciences(CIFMS,Nos.2022-I2M-2-002 and 2021-1-I2M-028).
文摘On-tissue chemical derivatization(OTCD)effectively enhances ionization efficiency of low abundant and poorly ionized functional molecules to improve detection sensitivity and coverage of mass spectrometry imaging(MSI).Combination OTCD and MSI provides a novel strategy for visualizing previously undisclosed metabolic heterogeneity in tumor.Herein,we present a method to visualize heterogeneous metabolism of oxylipins within tumor by coupling OTCD with airflow-assisted desorption electrospray ionization(AFADESI)-MSI.Taking Girard’s P as a derivatization reagent,easily ionized hydrazide and quaternary amine groups were introduced into the structure of carbonyl metabolites via condensation reaction.Oxylipins,including 127 fatty aldehydes(FALs)and 71 oxo fatty acids(FAs),were detected and imaged in esophageal cancer xenograft with AFADESI-MSI after OTCD.Then t-distributed stochastic neighbor embedding and random forest were exploited to precisely locate the distribution of oxylipins in heterogeneous tumor tissue.With this method,we surprisingly found almost all FALs and oxo FAs significantly accumulated in the core region of tumor,and exhibited a gradual increase trend in tumor over time.These results reveal spatiotemporal heterogeneity of oxylipins in tumor progression,highlighting the value of OTCD combined with MSI to gain deeper insights into understanding tumor metabolism.
基金supported by the National Natural Science Foundation of China(No.81803483,No.21927808)National Key Research and Development Program of China(No.2017YFC1704006)。
文摘Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy(DN)is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies.In the present study,a spatial-resolved metabolomics approach based on air flowassisted desorption electrospray ionization(AFADESI)and matrix-assisted laser desorption ionization(MALDI)integrated mass spectrometry imaging(MSI)was proposed to investigate tissue-specific metabolic alterations in the kidneys of high-fat diet-fed and streptozotocin(STZ)-treated DN rats and the therapeutic effect of astragalosideⅣ,a potential anti-diabetic drug,against DN.As a result,a wide range of functional metabolites including sugars,amino acids,nucleotides and their derivatives,fatty acids,phospholipids,sphingolipids,glycerides,carnitine and its derivatives,vitamins,peptides,and metal ions associated with DN were identified and their unique distribution patterns in the rat kidney were visualized with high chemical specificity and high spatial resolution.These region-specific metabolic disturbances were ameliorated by repeated oral administration of astragaloside Ⅳ(100 mg/kg)for 12 weeks.This study provided more comprehensive and detailed information about the tissue-specific metabolic reprogramming and molecular pathological signature in the kidney of diabetic rats.These findings highlighted the promising potential of AFADESI and MALDI integrated MSI based metabolomics approach for application in metabolic kidney diseases.
基金supported by the National Key Research and Development Program of China(No.2017YFC1704000)Outstanding Talent Support Program of Beijing,China(No.2017000020124G272)
文摘Understanding of the nephrotoxicity induced by drug candidates is vital to drug discovery and development.Herein,an in situ metabolomics method based on air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI)was established for direct analysis of metabolites in renal tissue sections.This method was subsequently applied to investigate spatially resolved metabolic profile changes in rat kidney after the administration of aristolochic acid I,a known nephrotoxic drug,aimed to discover metabolites associated with nephrotoxicity.As a result,38 metabolites related to the arginine-creatinine metabolic pathway,the urea cycle,the serine synthesis pathway,metabolism of lipids,choline,histamine,lysine,and adenosine triphosphate were significantly changed in the group treated with aristolochic acid I.These metabolites exhibited a unique distribution in rat kidney and a good spatial match with histopathological renal lesions.This study provides new insights into the mechanisms underlying aristolochic acids nephrotoxicity and demonstrates that AFADESI-MSI-based in situ metabolomics is a promising technique for investigation of the molecular mechanism of drug toxicity.
基金supported by Chinese Academy of Medical Science (CAMS) Innovation Fund for Medical Sciences (Grant Nos. 2016-I2M-1–009 and 2016-I2M-3–010)National Scientific and Technological Major Project for New Drugs (Grant No. 2017ZX09101003-002-004)
文摘With the rapid development and wide application of traditional Chinese medicine injection(TCMI), a number of adverse events of some TCMIs have incessantly been reported and have drawn broad attention in recent years. Establishing effective and practical analytical methods for safety evaluation and quality control of TCMI can help to improve the safety of TCMIs in clinical applications. In this study, a sensitive and rapid high-performance liquid chromatography–tandem mass spectrometry(HPLC–MS/MS)method has been developed and validated for the quantitative determination of potentially harmful substance5,5′-oxydimethylenebis(2-furfural, OMBF) in TCMI samples. Chromatographic separation was performed on a C18 reversed-phase column(150 mm × 2.1 mm, 5 μm) by gradient elution, using methanol–water containing 0.1% formic acid as mobile phase at the flow rate of 0.3 m L/min. MS/MS detection was performed on a triple quadrupole mass spectrometer with positive electrospray ionization in the multiple reaction-monitoring mode. The method was sensitive with a limit of quantification of 0.3 ng/m L and linear over the range of 0.3–30 ng/m L(r = 0.9998). Intra-and inter-day precision for analyte was o9.52% RSD withrecoveries in the range 88.0–109.67% at three concentration levels. The validated method was successfully applied to quantitatively determine the compound OMBF in TCMIs and glucose injections. Our study indicates that this method is simple, sensitive, practicable and reliable, and could be applied for safety evaluation and quality control of TCMIs and glucose injections.
基金supported by the National Instrumentation Program (No. 2016YFF0100304)the National Natural Science Foundation of China(Nos. 21335007, 81773678)+1 种基金the CAMS Innovation Fund for Medical Sciences(No. 2016-12 M-1-009)PUMC Youth Fund and the Fundamental Research Funds for the Central Universities(No. 3332015177)
文摘Mass spectrometry imaging (MSI) technology can simultaneously obtain the spatial distribution of thousands of chemical compounds and has unique advantages compared to other techniques that allow mapping the surface of bio-tissue. Here, we combined an air flow-assisted desorption electrospray ionization (AFADESI) MSI device with a high-resolution mass spectrometer to optimize the system parameters and achieve more accurate spatial distribution characteristics for compounds of interest while investigating bio-tissue sections. The platform set-up, required instrumentation, sample pretreatment, parameter optimization and bio-tissue characterization are described and discussed.Finally, the parameter conditions that can provide optimal ionic intensity and enhanced resolution were confirmed. The reasonable resolution and sensitivity improvements of AFADESI-MSI have been achieved through tandem a high-resolution mass spectrometer system, therefore, it would be a promising technique for the bio-tissue imaging analysis.
基金supported by CAMS Innovation Fund for Medical Sciences(No. 2016-12M-1-009)National Scientific and Technological Major Project for New Drugs(No.2017ZX09101003-002-004)
文摘A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry(UPLC–MS/MS) method was developed and fully validated for determination of arformoterol in rat plasma, lung and trachea tissues. The chromatographic separation was performed on an Agilent XDB C8 column with gradient elution by using acetonitrile and 0.1% formic acid water. The method presented high sensitivity(LLOQ of 1.83 pg/mL for plasma and 3.90 pg/mL for lung and trachea tissue homogenates) and great linearity over the range of 1.83–458 pg/mL for plasma, 13.9–1560 pg/mL for lung and trachea tissue homogenates. No carry over effect and matrix effect were observed. The intra-and inter-day precision/accuracy were within ±15% at three quality control concentration levels. This robust method was successfully applied to the pharmacokinetic, lung and trachea tissue distribution study after inhalation administration of arformoterol tartrate inhalation solution(50 m g/kg). The in vivo information indicate that arformoterol can be rapidly absorbed into blood through respiratory systems, lung and trachea tissue maintain a certain amount of arformoterol in 1 h after dosing.
文摘We have studied the fragmentation behavior of positive and negative ions of taxoi and 6/8/6 type taxoids, the influence of different substituents on fragmentation, and the correlativity between fragmentation patterns and structure by MS/MS technique with different ionization methods such as FAB-MS, ESI-MS, ete. We have also investigated in detail the fragmentation of various molecular-related ions, such as [M+H]+, [M+Na]+ and [M-H] ions, and the formation pathways of characteristic fragment ions. It has been found that there exists some competing reaction between the loss of C-13 side chain and decomposition by loss of acetic acid. In addition, by comparing CID spectra obtained with low- and high-energy collision, it is seen that CID-MS/MS with low-energy collision is more suitable for the study of the structural analysis of small molecules and drug metabolites. The experimental results demonstrate that MS/MS spectra can reflect more effectively the slight difference of structure between the related
基金financially supported by the National Instrumentation Program (No. 2011YQ17006702)the National Natural Science Foundation of China (No. 21027013 and No. 81102413)Fundamental Research Program of Shenzhen (No. JC201005280634A)
文摘The imaging mass spectrometry(IMS) technology has experienced a rapid development in recent years.A new IMS technology which is based on air flow assisted ionization(AFAI) was reported.It allows for the convenient pretreatment of the samples and can image a large area of sample in a single measurement with high sensitivity.The AFAI in DESI mode was used as the ion source in this paper.The new IMS method is named AFADESI-IMS.The adoption of assisted air flow makes the sample pretreatment easy and convenient.An optimization of the distance between the ion transport tube and MS orifice increases the sensitivity of the system.For data processing,a program based on MATLAB with the function of numerical analysis was developed.A theoretical imaging resolution of a few hundred microns can be achieved.The composite AFAI-IMS images of different target analytes were imaged with high sensitivity.A typical AFAI-IMS image of the whole-body section of a rat was obtained in a single analytical measurement.The ability to image a large area for relevant samples in a single measurement with high sensitivity and repeatability is a significant advantage.The method has enormous potentials in the MS imaging of large and complicated samples.
基金supported by the National Instrumentation Programmme(Nos.2011YQ17006702 and 2011YQ14015010)the National Natural Science Foundation of China(Nos.81102413 and 21175121)Fundamental Research Program of Shenzhen (No.JC201005280634A).
文摘A new multivariate statistical strategy for analyzing large datasets that are produced by imaging mass spectrometry(IMS) techniques is reported.The strategy divides the whole datacube of the sample into several subsets and analyses them one by one to obtain the results.Instead of analyzing the whole datacube at one time,the strategy makes the analysis easier and decreases the computation time greatly.In this report,the IMS data are produced by the air flow-assisted ionization IMS(AFAI-IMS).The strategy can be used in combination with most multivariate statistical analysis methods.In this paper,the strategy was combined with the principal component analysis(PCA) and partial least square analysis(PLS).It was proven to be effective by analyzing the handwriting sample.By using the strategy,the m/z corresponding to the specific lipids in rat brain tissue were distinguished successfully.Moreover the analysis time grew linearly instead of exponentially as the size of sample increased.The strategy developed in this study has enormous potential for searching for the mjz of potential biomarkers quickly and effectively.
文摘A ’H NMR signal shifted drastically to down field (δ - 10.0) at the bay area and in dose proximity to C = O group for H-1 was observed through complete assignments of 1H NMR spectra for pyridino- and benzobenzanthrones. It is concluded that this phenomenon is due not only to the anisotropy effect of C=O plus aromatic ring current effect, but also to the electrostatic attraction of C-H (δ+)…O(δ-)=C interaction. The evidence for the sp2C-H…O = C intramolecular interaction similar to hydrogen bond has also been given by EI-MS and MS/MS (CID) spectroscopy and IR spectroscopy, as well as MNDO-PM3 calculations. This new kind of interaction might be called ’quasi-hydrogen bond’.