scholarly journals Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2195
Author(s):  
Yasushi Ueyanagi ◽  
Daiki Setoyama ◽  
Daisuke Kawakami ◽  
Yuichi Mushimoto ◽  
Shinya Matsumoto ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Organic Acids ◽  
Multiple Reaction Monitoring ◽  
Automated System ◽  
Negative Ion ◽  
Automation System ◽  
Proof Of Concept ◽  
Hospital System ◽  
Organic Acidemias ◽  
Concept Validation

Gas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to develop a fully automated system for quantifying serum organic acids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The pretreatment CLAM-2030 device was connected to an LC-MS/MS system for processing serum under optimized conditions, which included derivatizing serum organic acids using 3-Nitrophenylhydrazine. The derivatized organic acids were separated on a reverse-phase Sceptor HD-C column and detected using negative-ion electrospray ionization multiple reaction monitoring MS. The automated pretreatment-LC-MS/MS system processed serum in less than 1 h and analyzed 19 serum organic acids, which are used to detect organic acidemias. The system exhibited high quantitative sensitivity ranging from approximately 2 to 100 µM with a measurement reproducibility of 10.4% CV. Moreover, a proof-of-concept validation of the system was performed using sera from patients with propionic acidemia (n = 5), methylmalonic acidemia (n = 2), and 3-methylcrotonylglycinuria (n = 1). The levels of marker organic acids specific to each disease were significantly elevated in the sera of the patients compared to those in control samples. The automated pretreatment-LC-MS/MS system can be used as a rapid in-hospital system to measure organic acid levels in serum for the diagnosis of organic acidemias.

scholarly journals Comprehensive Screening of Urine Samples for Inborn Errors of Metabolism by Electrospray Tandem Mass Spectrometry

2002 ◽  
Vol 48 (11) ◽  
pp. 1970-1980 ◽  
Author(s):  
James J Pitt ◽  
Mary Eggington ◽  
Stephen G Kahler
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Inborn Errors Of Metabolism ◽  
Multiple Reaction Monitoring ◽  
Direct Injection ◽  
Negative Ion ◽  
Screening Methods ◽  
Tandem Mass ◽  
Inborn Errors ◽  
Ion Analysis

Abstract Background: Detection of abnormal metabolites in urine is important for the diagnosis of many inborn errors of metabolism (IEM). Rapid, comprehensive screening methods are needed. Methods: We used electrospray ionization tandem mass spectrometry in positive- and negative-ion modes to detect selected metabolites in urine. For positive-ion analysis, samples were dried and butylated, whereas for negative-ion analysis, samples were merely diluted with the mobile phase. Analysis was by direct injection with multiple reaction monitoring for 32 metabolites in positive mode (amino acids and acylcarnitines) and 30 metabolites in negative mode (organic acids). Run time was 2.1 min in each mode. Results: Interbatch CVs ranged from 4.8% to 32%, enabling quantification of many metabolites. The procedure was applied to controls (278 and 120 in positive- and negative-ion mode, respectively) and 108 IEM individuals representing 37 different IEM. In 105 IEM individuals, representing 36 different IEM, concentrations of one or more diagnostic metabolites were above the 99th percentiles of the control values. Conclusions: The procedure is faster and less labor-intensive than conventional methods of testing for IEM by amino and organic acid profiling and has similar diagnostic sensitivity. The ability to include a greater range of metabolites offers the potential of a more comprehensive screening procedure.

scholarly journals Detection and Identification of Zeranol in Chicken or Rabbit Liver by Liquid Chromatography-Electrospray Tandem Mass Spectrometry

2002 ◽  
Vol 85 (4) ◽  
pp. 841-847 ◽  
Author(s):  
Xiaoming Fang ◽  
Jiahua Chen ◽  
Dehua Guo ◽  
Guoquan Wang
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Multiple Reaction Monitoring ◽  
Rabbit Liver ◽  
Negative Ion ◽  
Tandem Mass ◽  
Detection And Identification ◽  
Relative Standard ◽  
Limit Of Quantitation

Abstract A sensitive, specific, and reliable liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for detection and identification of zeranol in chicken or rabbit liver. A homogenized liver sample was hydrolyzed with β-glucuronidase/arylsulfatase, and the hydrolysate was extracted with ethyl ether. The supernatant was evaporated to dryness, and the residue was dissolved in chloroform and re-extracted with sodium hydroxide. After acidification, the extract was cleaned up on a C18 solid-phase extraction cartridge and analyzed by electrospray LC-MS/MS in the negative ion mode. The multiple reaction monitoring transition from both m/z 321 to 277 and m/z 321 to 303 was monitored for confirmation, and the product ion of 277 was used for quantitation. Separation was performed on a Waters XTettra™ C18 column (50 × 2.1 mm, 3.5 μm) combined with a safeguard column (Symmetry C18, 20 × 3.9 mm, 5 μm), using a gradient elution with acetonitrile and 20mM ammonium acetate. Calibration curves were prepared and good linearity was achieved over the concentration ranges tested. For all liver samples fortified at 3 different levels of 1, 5, and 50 μg/kg, the overall recoveries and relative standard deviations were in the range of 61–90 and 8–13%, respectively. The limit of quantitation based on the assay validation was 1 μg/kg. The method had been used on a routine basis for detection and identification of zeranol in liver samples.

scholarly journals Development and application of a liquid chromatography-mass spectrometry method for the determination of sugars and organics acids in araza, ceriguela, guava, mango and pitanga

2021 ◽  
Vol 24 ◽  
Author(s):  
Patricia Carvalho Cardoso ◽  
Fernanda Sviech ◽  
Marcella Fernanda Alves Reis ◽  
Amadeu Hoshi Iglesias ◽  
Rafael Augustus Oliveira ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Product Development ◽  
Organic Acids ◽  
Multiple Reaction Monitoring ◽  
Psidium Guajava ◽  
Ionization Mass ◽  
Tropical Fruits ◽  
Esi Ms ◽  
Spondias Purpurea

Abstract The intrinsic characteristics of many tropical fruits cause high post-harvest losses and prevent their commercialization as fresh fruits. Information about their composition is crucial for defining processing conditions and identifying opportunities for product development. However, the analytical methods generally used to quantify sugars and organic acids are costly and time-consuming. Simultaneous analysis by Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (LC-ESI-MS/MS) is a very sensitive and reproducible technique, allowing for accurate simultaneous multi-analyte quantitation in complex systems. Thus, a LC-ESI-MS/MS Multiple Reaction Monitoring (MRM) method was developed using reverse phase column for detecting and quantifying sugars and some organic acids in only 4 min, in selected fruits as following: araza (Psidium cattleianum L.), ciriguela (Spondias purpurea L.), mango (Mangifera indicaL.), guava (Psidium guajava L.) and pitanga (Eugenia uniflora L.). All fruits had a similar concentration of glucose and fructose, except for pitanga that presented higher values of both. The content of citric, malic and tartaric acids was quantitated; some fruits stood out by their high content of organic acids, for instance, araza was rich in citric acid. The newly generated data on the composition of tropical fruits allowed establishing a correlation between the carbohydrate content and the physical properties of the fruit pulps, assist in product development.

scholarly journals Simultaneous determination of multi – component mycotoxin in feeds by liquid chromatography – tandem mass spectrometry

2013 ◽  
Vol 57 (4) ◽  
pp. 567-572 ◽  
Author(s):  
Katarzyna Pietruszka ◽  
Bartosz Sell ◽  
Olga Burek ◽  
Henryka Wiśniewska-Dmytrow
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Ion Trap ◽  
Limit Of Detection ◽  
Multiple Reaction Monitoring ◽  
Negative Ion ◽  
Tandem Mass ◽  
Penicillium Species ◽  
Liquid Chromatography Tandem Mass

Abstract A multiresidue method for determination and quantification of Fusarium mycotoxins: deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin, and metabolite of Aspergillus and Penicillium species - ochratoxin A in feeds was described. The method was based on the simultaneous extraction of selected mycotoxins from matrix, followed by liquid chromatography coupled with tandem mass spectrometry using a hybrid triple quadrupole - linear ion trap mass spectrometer with the multiple reaction monitoring in both positive- and negative-ion modes. The method was validated in accordance with the Commision Decision 2002/657/EC requirements. The mean recoveries of mycotoxins from spiked feed samples ranged from 74.6% to 113.9%, whereas limit of detection and quantification ranged from 0.72 to 12.4 μg/kg and 1.86 to 31.7 μg/kg, respectively.

Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases

2020 ◽  
Vol 58 (5) ◽  
pp. 787-797 ◽  
Author(s):  
Xiaowei Fu ◽  
Yi Xiao ◽  
Jamie Golden ◽  
Sizhe Niu ◽  
Christopher P. Gayer
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Bile Acids ◽  
Multiple Reaction Monitoring ◽  
Negative Ion ◽  
Tandem Mass ◽  
Reaction Monitoring ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

AbstractBackgroundA method for bile acid profiling measuring 21 primary and secondary bile acids in serum samples was developed and validated with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample preparation included spiking with internal standards followed by protein precipitation, centrifugation, drying under nitrogen gas and reconstitution. Extracted samples were injected onto a Phenomenex Kinetex C18 column (150 × 4.60 mm, 2.6 μm).MethodsData was collected with LC-MS/MS operated in negative ion mode with multiple reaction monitoring (MRM) and single reaction monitoring (SRM). The analytical run time was 12 min.ResultsThe method showed excellent linearity with high regression coefficients (>0.99) over a range of 0.05 and 25 μM for all analytes tested. The method also showed acceptable intra-day and inter-day accuracy and precision. As a proof of concept, the analytical method was applied to patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), biliary atresia (BA), and necrotizing enterocolitis (NEC), and distinct bile acids profiles were demonstrated.ConclusionsThe method could be poised to identify possible biomarkers for non-invasive early diagnosis of these disorders.

scholarly journals Electrospray tandem mass spectrometric measurements of organotin compounds

2004 ◽  
Vol 18 (1) ◽  
pp. 95-112 ◽  
Author(s):  
Joseph H. Banoub ◽  
Judith Miller-Banoub ◽  
George V. Sheppard ◽  
Howard J. Hodder
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Reference Materials ◽  
Multiple Reaction Monitoring ◽  
Low Energy ◽  
Organotin Compounds ◽  
Negative Ion ◽  
Tandem Mass ◽  
Environmental Matrices ◽  
Tandem Mass Spectrometric

Electrospray mass spectrometry of a series of organotin compounds in solutions of methanol are reported. Low energy collision‒induced dissociation MS/MS analysis of diagnostic precursor ions confirmed the characteristic fingerprint patterns obtained in the conventional electrospray spectra and proved to be a specific and very sensitive method for quantification of the (R3Sn)2O and the series of RnSnX4–ncompounds in environmental matrices. Concentrations of butyltin compounds (TBTX, DBTX2and MBTX3) in sediment reference materials PACS-1 and PACS-2 and butyltin and phenyltin compounds (TBTX, DBTX2, MBTX3, TPTX, DPTX2and DPTX3) in Quasimeme II biota reference material (QSP001BT) were determined. The organotin compounds were extracted from the reference materials with 1-butanol followed by dilution with methanol containing 1 mM ammonium acetate. The extracts were introduced directly into the electrospray source by a continuous flow of MeOH : H2O (60 :40). Quantitation of TBTX, DBTX2, TPTX, DPTX2and DPTX3was achieved by low energy CID tandem mass spectrometry using the Multiple Reaction Monitoring (MRM) analysis with the appropriate MS/MS transitions (positive ion electrospray ionization). Quantitation of MBTX3was achieved using a negative ion electrospray CID tandem mass spectrometry method. For all samples quantitation was achieved by use of the method of standard addition, relative extraction recoveries were determined spiking with internal standards of mono‒, di‒ and triorganotin compounds separately to different samples.

Confounding contaminants in mass spectrometric reaction monitoring

2018 ◽  
Author(s):  
Gilian T. Thomas ◽  
Landon MacGillivray ◽  
Natalie L. Dean ◽  
Rhonda L. Stoddard ◽  
Lars Yunker ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectrometric ◽  
Negative Ion ◽  
Reaction Monitoring ◽  
Schlenk Flask ◽  
Dynamic Analyses ◽  
Rubber Septa ◽  
Negative Ion Mode ◽  
Mode A

<p>Reactions carried out in the presence of rubber septa run the risk of additives being leached out by the solvent. Normally, such species are present at low enough levels that they do not interfere with the reaction significantly. However, when studying reactions using sensitive methods such as mass spectrometry, the appearance of even trace amounts of material can confuse dynamic analyses of reactions. A wide variety of additives are present in rubber along with the polymer: antioxidants, dyes, detergent, and vulcanization agents, and these are all especially problematic in negative ion mode. A redesigned Schlenk flask for pressurized sample infusion (PSI) is presented as a means of practically eliminating the presence of contaminants during reaction analyses.</p>

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