A simple method was developed for rapid and direct profiling of alkaloids in medical herbs via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. The dry herbs were first gro...A simple method was developed for rapid and direct profiling of alkaloids in medical herbs via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. The dry herbs were first ground to powder and passed through a stainless steel sieve, mixed with the matrix solution to form a homogeneous suspension, which was then directly applied to MALDI analysis. Several matrices were investigated and 2,5- dihydroxybenzoic acid（DHB） was chosen as the optimized one, and the particle with small size was found to favor the analysis. Using this method, the profiles of alkaloids in several medical herbs were readily obtained, and the toxicities of crude and processed Radix Aconiti Lateralis Preparata were compared via the relative intensities of the peaks of the corresponding toxic components shown in their MALDI spectra. This method therefore provides a rapid and reliable protocol for obtaining profiles of alkaloids in medical herbs by using MALDI-TOF MS.展开更多
Introduction Single nucleotide polymorphisms （SNPs） are the most abundant DNA markers in the human genome occurring at a frequency of one in every 500--1000 nucleotides. A variety of methods have been used for the ...Introduction Single nucleotide polymorphisms （SNPs） are the most abundant DNA markers in the human genome occurring at a frequency of one in every 500--1000 nucleotides. A variety of methods have been used for the analysis of single nucleotide polymorphisms, including restriction fragment length polymorphism （RFLP）, direct sequencing by using laser-induced fluorescence detectionTM, fluorescence energy transfer, MALDI-TOF MS combined with primer extension or invasive cleavage, and fluorescence polarization. During the past two decades, mass spectrometry has become a very popular tool in the analysis of biomolecules and is perfectly suited to the analysis of single nucleotide polymorphisms （SNPs） due to its speed, low cost, and accuracy. In this work, we used MALDI TOF mass spectrometry to detect the fragments of restriction endonuclease hydrolysis of PCR products flanking a SNP located at paraoxonase 1（Q192R）. Compared with electrophoresis, this method requires less time of analysis and possess a higher accuracy.展开更多
Matrix-assisted Laser Desorption/Ionization with Time-of-flight Mass Spectrometry (MALDI-TOFMS) was investigated as a method for the rapid identifica-tion of species. Current demand in microbial identi-fication is how...Matrix-assisted Laser Desorption/Ionization with Time-of-flight Mass Spectrometry (MALDI-TOFMS) was investigated as a method for the rapid identifica-tion of species. Current demand in microbial identi-fication is how to compare unknown strains to the known one quickly, semi-automatically and accurately. In this paper, we present a software tool that allows flexibly microbial matching in a user-friendly way, by letting the users to customize comparison parameters including: in vitro transcription enzyme, mass tolerance,minimum fragment length, intensity threshold and corresponding weights. We provide three spectral scoring functions to compute the affin-ity between the species. Therefore, the precision of microbial comparison increases. To test and verify this tool, we employed experimental spectral data based on MALDI-TOFMS and the gene sequences of E.coli and Salmonella. This software is written in Java for cross-platform intention.展开更多
目的:研究比较两种基质辅助激光解吸电离飞行时间质谱仪(Autof ms 1000与MALDI Biotyper)对临床常见丝状病原真菌鉴定的准确性和效率。方法:共纳入32株丝状病原真菌标准菌株和120株临床分离菌株,按照标准操作方法分别用Autof ms 1000与M...目的:研究比较两种基质辅助激光解吸电离飞行时间质谱仪(Autof ms 1000与MALDI Biotyper)对临床常见丝状病原真菌鉴定的准确性和效率。方法:共纳入32株丝状病原真菌标准菌株和120株临床分离菌株,按照标准操作方法分别用Autof ms 1000与MALDI Biotyper对相关菌株进行鉴定,对相关结果及测序分子鉴定结果进行对比分析。结果:在种水平上,Autof ms 1000质谱仪共检出140株丝状病原真菌,检出正确率为92.1%(140/152),其中126株分值在9.0以上,2株出现错误鉴定,误鉴定比例为1.3%(2/152);MALDI Biotyper质谱仪共正确检出98株,检出正确率为64.4%,2.0分以上的菌株47株,仅3株鉴定分值大于2.3分,7株出现误鉴定。结论:Autof ms 1000与MALDI Biotyper这两种质谱鉴定系统均能用于临床常见丝状病原真菌的快速鉴定,而Autof ms 1000质谱仪更具优势。展开更多
目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)结合短时培养在快速鉴定血流感染病原菌中的应用。方法对南京医科大学附属淮安第一医院微生物...目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)结合短时培养在快速鉴定血流感染病原菌中的应用。方法对南京医科大学附属淮安第一医院微生物室2020年11月至2021年12月血培养报告阳性的标本进行革兰染色分类,并采用4h和6h短时培养法及MALDI-TOF MS技术快速鉴定感染病原菌并实时报送病区单元。结果4h短时培养革兰阴性菌共351株,339株(96.6%)细菌鉴定到种属,与24h鉴定结果相同,11株无鉴定结果,1株鉴定错误;6h短时培养革兰阳性菌共216株,212株(98.1%)鉴定到种属,与24h鉴定结果相同,3株无鉴定结果,1株鉴定错误。跟踪321株革兰阴性菌的临床反馈,287例(89.4%)给予临床关注,89例(27.7%)进行药物调整,238例(74.1%)临床疗效良好;跟踪183株革兰阳性菌的临床反馈,其中150例(82.0%)给予临床关注,49例(26.8%)进行药物调整,131例(71.6%)临床疗效良好。结论MALDI-TOF MS技术结合短时培养方法可有效提高血流感染病原菌鉴定准确率,改善血流感染患者抗菌药物治疗时机。展开更多
Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser des...Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) to investigate the spatial distribution of endogenous compounds(e.g., lipids) in CCS. However, only limited lipid species can be detected owing to a low ion yield by using MALDI. Besides, it is still challenging to fully characterize the structural diversity of lipids due to the existence of isomeric/isobaric species. Here, we carried out the initial application of MALDI coupled with laser-postionization(MALDI-2) and trapped ion mobility spectrometry(TIMS) imaging in HCT116 colon CCS to address these challenges. We demonstrated that MALDI-2 is capable of detecting more number and classes of lipids in HCT116 colon CCS with higher signal intensities than MALDI. TIMS could successfully separate numerous isobaric/isomeric species of lipids in CCS. Interestingly, we found that some isomeric/isobaric species have totally different spatial distributions in colon CCS. Further MS/MS imaging analysis was employed to determine the compositions of fatty acid chains for isomeric species by examining disparities in signal intensities and spatial distributions of product ions. This work stresses the robust ability of TIMS and MALDI-2 imaging in analyzing endogenous lipids in CCS, which could potentially become powerful tools for future cancer studies.展开更多
[目的]建立基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)技术检测不同禽肉动物源性成分的方法,构建不同禽肉蛋白质分子指纹图谱标准数据库,对不同...[目的]建立基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)技术检测不同禽肉动物源性成分的方法,构建不同禽肉蛋白质分子指纹图谱标准数据库,对不同禽肉进行鉴别。[方法]样品用磷酸盐缓冲液提取,净化后用MALDI-TOF MS法测定。采用ChemPattern软件对不同禽肉蛋白图谱进行化学计量学分析,建立识别模型。[结果]鸡、鸭、鹅3种不同禽肉之间具有明显差异。首次建立不同禽肉蛋白质分子指纹图谱标准数据库,可一次性对每个样品中鸡、鸭、鹅等肉源性成分进行同时分析。[结论]建立的基于蛋白质组学的鸡、鸭、鹅源性成分鉴定系统具有简便、稳定、准确、重现性好的优点,适用于不同禽肉中鸡鸭鹅动物源性成分的鉴别。展开更多
基金Supported by the Major State Basic Research Development Program of China(No.2006CB5047060)the National Natural Science Foundation of China(Nos.30672600, 30772721)the Natural Science Foundation of Jilin Province, China (No.20060902)
文摘A simple method was developed for rapid and direct profiling of alkaloids in medical herbs via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. The dry herbs were first ground to powder and passed through a stainless steel sieve, mixed with the matrix solution to form a homogeneous suspension, which was then directly applied to MALDI analysis. Several matrices were investigated and 2,5- dihydroxybenzoic acid（DHB） was chosen as the optimized one, and the particle with small size was found to favor the analysis. Using this method, the profiles of alkaloids in several medical herbs were readily obtained, and the toxicities of crude and processed Radix Aconiti Lateralis Preparata were compared via the relative intensities of the peaks of the corresponding toxic components shown in their MALDI spectra. This method therefore provides a rapid and reliable protocol for obtaining profiles of alkaloids in medical herbs by using MALDI-TOF MS.
文摘Introduction Single nucleotide polymorphisms （SNPs） are the most abundant DNA markers in the human genome occurring at a frequency of one in every 500--1000 nucleotides. A variety of methods have been used for the analysis of single nucleotide polymorphisms, including restriction fragment length polymorphism （RFLP）, direct sequencing by using laser-induced fluorescence detectionTM, fluorescence energy transfer, MALDI-TOF MS combined with primer extension or invasive cleavage, and fluorescence polarization. During the past two decades, mass spectrometry has become a very popular tool in the analysis of biomolecules and is perfectly suited to the analysis of single nucleotide polymorphisms （SNPs） due to its speed, low cost, and accuracy. In this work, we used MALDI TOF mass spectrometry to detect the fragments of restriction endonuclease hydrolysis of PCR products flanking a SNP located at paraoxonase 1（Q192R）. Compared with electrophoresis, this method requires less time of analysis and possess a higher accuracy.
文摘Matrix-assisted Laser Desorption/Ionization with Time-of-flight Mass Spectrometry (MALDI-TOFMS) was investigated as a method for the rapid identifica-tion of species. Current demand in microbial identi-fication is how to compare unknown strains to the known one quickly, semi-automatically and accurately. In this paper, we present a software tool that allows flexibly microbial matching in a user-friendly way, by letting the users to customize comparison parameters including: in vitro transcription enzyme, mass tolerance,minimum fragment length, intensity threshold and corresponding weights. We provide three spectral scoring functions to compute the affin-ity between the species. Therefore, the precision of microbial comparison increases. To test and verify this tool, we employed experimental spectral data based on MALDI-TOFMS and the gene sequences of E.coli and Salmonella. This software is written in Java for cross-platform intention.
文摘目的:研究比较两种基质辅助激光解吸电离飞行时间质谱仪(Autof ms 1000与MALDI Biotyper)对临床常见丝状病原真菌鉴定的准确性和效率。方法:共纳入32株丝状病原真菌标准菌株和120株临床分离菌株,按照标准操作方法分别用Autof ms 1000与MALDI Biotyper对相关菌株进行鉴定,对相关结果及测序分子鉴定结果进行对比分析。结果:在种水平上,Autof ms 1000质谱仪共检出140株丝状病原真菌,检出正确率为92.1%(140/152),其中126株分值在9.0以上,2株出现错误鉴定,误鉴定比例为1.3%(2/152);MALDI Biotyper质谱仪共正确检出98株,检出正确率为64.4%,2.0分以上的菌株47株,仅3株鉴定分值大于2.3分,7株出现误鉴定。结论:Autof ms 1000与MALDI Biotyper这两种质谱鉴定系统均能用于临床常见丝状病原真菌的快速鉴定,而Autof ms 1000质谱仪更具优势。
文摘目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)结合短时培养在快速鉴定血流感染病原菌中的应用。方法对南京医科大学附属淮安第一医院微生物室2020年11月至2021年12月血培养报告阳性的标本进行革兰染色分类,并采用4h和6h短时培养法及MALDI-TOF MS技术快速鉴定感染病原菌并实时报送病区单元。结果4h短时培养革兰阴性菌共351株,339株(96.6%)细菌鉴定到种属,与24h鉴定结果相同,11株无鉴定结果,1株鉴定错误;6h短时培养革兰阳性菌共216株,212株(98.1%)鉴定到种属,与24h鉴定结果相同,3株无鉴定结果,1株鉴定错误。跟踪321株革兰阴性菌的临床反馈,287例(89.4%)给予临床关注,89例(27.7%)进行药物调整,238例(74.1%)临床疗效良好;跟踪183株革兰阳性菌的临床反馈,其中150例(82.0%)给予临床关注,49例(26.8%)进行药物调整,131例(71.6%)临床疗效良好。结论MALDI-TOF MS技术结合短时培养方法可有效提高血流感染病原菌鉴定准确率,改善血流感染患者抗菌药物治疗时机。
基金supported by the National Natural Science Foundation of China (Nos.22036001, 22276034 and 22106130)。
文摘Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) to investigate the spatial distribution of endogenous compounds(e.g., lipids) in CCS. However, only limited lipid species can be detected owing to a low ion yield by using MALDI. Besides, it is still challenging to fully characterize the structural diversity of lipids due to the existence of isomeric/isobaric species. Here, we carried out the initial application of MALDI coupled with laser-postionization(MALDI-2) and trapped ion mobility spectrometry(TIMS) imaging in HCT116 colon CCS to address these challenges. We demonstrated that MALDI-2 is capable of detecting more number and classes of lipids in HCT116 colon CCS with higher signal intensities than MALDI. TIMS could successfully separate numerous isobaric/isomeric species of lipids in CCS. Interestingly, we found that some isomeric/isobaric species have totally different spatial distributions in colon CCS. Further MS/MS imaging analysis was employed to determine the compositions of fatty acid chains for isomeric species by examining disparities in signal intensities and spatial distributions of product ions. This work stresses the robust ability of TIMS and MALDI-2 imaging in analyzing endogenous lipids in CCS, which could potentially become powerful tools for future cancer studies.
