scholarly journals Comprehensive Detection of Disorders of Purine and Pyrimidine Metabolism by HPLC with Electrospray Ionization Tandem Mass Spectrometry

2006 ◽  
Vol 52 (6) ◽  
pp. 1127-1137 ◽  
Author(s):  
Susen Hartmann ◽  
Jürgen G Okun ◽  
Christiane Schmidt ◽  
Claus-Dieter Langhans ◽  
Sven F Garbade ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Multiple Reaction Monitoring ◽  
Dihydropyrimidine Dehydrogenase ◽  
Urine Samples ◽  
Tandem Mass ◽  
Pyrimidine Metabolism ◽  
Phosphoribosyl Transferase ◽  
Ionization Tandem Mass Spectrometry

Abstract Background: Clinical presentation and disease severity in disorders of purine and pyrimidine metabolism vary considerably. We present a method that allows comprehensive, sensitive, and specific diagnosis of the entire spectrum of abnormalities in purine and pyrimidine metabolism in 1 analytical run. Methods: We used reversed-phase HPLC electrospray ionization tandem mass spectrometry to investigate 24 metabolites of purine and pyrimidine metabolism in urine samples from healthy persons and from patients with confirmed diagnoses of inherited metabolic disorders. Urine samples were filtered and diluted to a creatinine concentration of 0.5 mmol/L. Stable-isotope–labeled internal standards were used for quantification. The metabolites were analyzed by multiple-reaction monitoring in positive and negative ionization modes. Results: Total time of analysis was 20 min. Recovery (n = 8) of a compound after addition of a known concentration was 85%–133%. The mean intraday variation (n = 10) was 12%. The interday variation (n = 7) was ≤17%. Age-related reference intervals were established for each compound. Analysis of patient urine samples revealed major differences in tandem mass spectrometry profiles compared with those of control samples. Twelve deficiencies were reliably detected: hypoxanthine guanine phosphoribosyl transferase, xanthine dehydrogenase, purine nucleoside phosphorylase, adenylosuccinate lyase, uridine monophosphate synthase, adenosine deaminase, adenine phosphoribosyl transferase, molybdenum cofactor, thymidine phosphorylase, dihydropyrimidine dehydrogenase, dihydropyrimidinase, and β-ureidopropionase. Conclusion: This method enables reliable detection of 13 defects in purine and pyrimidine metabolism in a single analytical run.

scholarly journals Defects in Pyrimidine Degradation Identified by HPLC-Electrospray Tandem Mass Spectrometry of Urine Specimens or Urine-soaked Filter Paper Strips

2000 ◽  
Vol 46 (12) ◽  
pp. 1916-1922 ◽  
Author(s):  
Henk van Lenthe ◽  
André B P van Kuilenburg ◽  
Tetsuya Ito ◽  
Albert H Bootsma ◽  
Arno van Cruchten ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Filter Paper ◽  
Degradation Products ◽  
Degradation Pathway ◽  
Urine Samples ◽  
Tandem Mass ◽  
Pyrimidine Degradation ◽  
Ionization Tandem Mass Spectrometry

Abstract Background: Urinary concentrations of thymine, uracil, and their degradation products are useful indicators of deficiencies of enzymes of the pyrimidine degradation pathway. We describe a rapid, specific method to measure these concentrations to detect inborn errors of pyrimidine metabolism. Methods: We used urine or urine-soaked filter-paper strips as samples and measured thymine, uracil, and their degradation products dihydrothymine, dihydrouracil, N-carbamyl-β-aminoisobutyric acid, and N-carbamyl-β-alanine. Reversed-phase HPLC was combined with electrospray ionization tandem mass spectrometry, and detection was performed by multiple-reaction monitoring. Stable-isotope-labeled reference compounds were used as internal standards. Results: All pyrimidine degradation products could be measured in one analytical run of 15 min. Detection limits were 0.4–4 μmol/L. The intraassay imprecision (CV) of urine samples with added compounds was 1.3–12% for liquid urines and 1.0–10% for filter-paper extracts of the urines. The interassay imprecision (CV) was 3–11% (100–200 μmol/L). Recoveries were 89–99% at 100–200 μmol/L and 95–106% at 1 mmol/L in liquid urines, and 93–103% at 100–200 μmol/L and 100–106% at 1 mmol/L in filter-paper samples. Correct identifications of deficiencies of the pyrimidine-degrading enzymes were readily made with urine samples from patients with known defects. Conclusions: HPLC with electrospray ionization tandem mass spectrometry allows rapid testing for disorders of the pyrimidine degradation pathway, and filter-paper samples allow easy collection, transport, and storage of urine samples.

scholarly journals Differentiation between 3,4- and 4,15-Epoxyeudesmanolides by Electrospray Ionization Tandem Mass Spectrometry

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Herbert Júnior Dias ◽  
Ricardo Stefani ◽  
José Carlos Tomaz ◽  
Ricardo Vessecchi ◽  
Antônio Eduardo Miller Crotti
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Sesquiterpene Lactones ◽  
Multiple Reaction Monitoring ◽  
Lone Pair ◽  
Fragmentation Pathway ◽  
Tandem Mass ◽  
Product Ions ◽  
Ionization Tandem Mass Spectrometry

We investigated the fragmentation of the eudesmanolide-type sesquiterpene lactones 1α-(4-hydroxymethacryloyloxy)-3α,4α-epoxy-8α-hydroxyeudesm-11(13)-6α,12-olide (1) and 1α-(2,3-epoxyangeloyloxy)-4α,15-epoxy-8α-hydroxyeudesm-11(13)-6α,12-olide (2) by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The elimination of the different ester substituent at C(1) directly from protonated 1 and 2 (A) led to the formation of two regioisomer product ions B (A − RCO2H). Further fragmentation of B resulted from consecutive eliminations of H2O and CO molecules. However, we identified four product ions that allowed for the differentiation between 3,4- and 4,15-epoxyeudesmanolides. The formation of these diagnostic ions was associated with the C(3)–O bond of compound 1, which propitiates the participation of the lone pair of the oxygen epoxide in the formation of B through a Grob-Wharton-type fragmentation, then resulting in an alternative fragmentation pathway. These data can be useful for the fast differentiation between epoxyeudesmanolide regioisomers directly from Dimerostemma extracts by liquid chromatography-tandem mass spectrometry (LC-MS/MS), as an alternative to NMR, or even for quantitation studies of these compounds using multiple reaction monitoring (MRM) scan.

scholarly journals Confirmatory Determination of Six Penicillins in Honey by Liquid Chromatography/Electrospray Ionization–Tandem Mass Spectrometry

2004 ◽  
Vol 87 (1) ◽  
pp. 45-55 ◽  
Author(s):  
Jian Wang
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Solid Phase ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Penicillin G ◽  
Tandem Mass ◽  
Ionization Tandem Mass Spectrometry

Abstract A confirmatory method for 6 penicillin antibiotics (amoxicillin, ampicillin, penicillin G, oxacillin, cloxacillin, and dicloxacillin) in honey is presented that allows determination and confirmation of identity of the antibiotics at trace levels. The method includes the use of a stable isotope-labeled internal standard benzyl (d7-phenyl) penicillate and removal of sugar and other substances by solvent and solid-phase extraction. The honey extracts are then analyzed for penicillin residues by liquid chromatography/electrospray ionization–tandem mass spectrometry. Mass spectral acquisition was achieved in an electrospray positive ion mode by applying multiple reaction monitoring of 2 or 3 fragment ion transitions to provide a high degree of sensitivity and specificity. Typical recoveries of 6 penicillins at fortification levels of 6, 16, 40, and 80 μg/kg ranged from 51.4 to 132.9%. The recoveries varied with the individual penicillins and were affected by different honey matrixes. The ion ratios were consistent and could be used for confirmation of identity of the penicillins. The method limits of detection (μg/kg) were 0.25 for amoxicillin, 0.19 for ampicillin, 0.068 for penicillin G, 0.028 for oxacillin, 0.052 for cloxacillin, and 0.085 for dicloxacillin. The method limits of confirmation (μg/kg) were 0.44 for amoxicillin, 0.52 for ampicillin, 0.23 for penicillin G, 0.14 for oxacillin, 0.14 for cloxacillin, and 0.15 for dicloxacillin when a sample size of 5 g honey was used.

scholarly journals Determination of Pesticides in Apples by Liquid Chromatography with Electrospray Ionization Tandem Mass Spectrometry and Estimation of Measurement Uncertainty

2007 ◽  
Vol 90 (2) ◽  
pp. 550-567 ◽  
Author(s):  
Jian Wang ◽  
David Wotherspoon
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Measurement Uncertainty ◽  
Electrospray Ionization ◽  
Signal To Noise Ratio ◽  
Multiple Reaction Monitoring ◽  
Tandem Mass ◽  
Intermediate Precision ◽  
Ionization Tandem Mass Spectrometry

Abstract A liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) method was developed and validated to determine 90 pesticides in apple samples. MS/MS data acquisition was achieved by applying multiple-reaction monitoring of 2 fragment ion transitions to provide a high degree of sensitivity and selectivity for both quantitation and confirmation. Matrix-matched standard calibration curves with the use of isotopically labeled standards (or a chemical analog) as internal standards were used to achieve the best accuracy for the method. Both a conventional method validation procedure and a designed experiment were applied to study the accuracy and precision of the method. A compiled computer program that provided a semiautomated procedure for handling a large number of calculations was used to calculate the overall recovery, intermediate precision, and measurement uncertainty (MU). In general, the overall recoveries from samples spiked at levels of 10, 50, and 80 g/kg, ranged from 60.8 to 121.1%, intermediate precision was <10%, and MUs were <30%. Poor accuracy and/or repeatability was observed for pyridate and etofenprox. The method limits of detection based on a signal-to-noise ratio of >3 were usually <1 g/kg (ppb), except those for aldicarb sulfoxide and pyridaphenthion, which were about 5 g/kg.

scholarly journals Analysis of Narrow-Leaf Lupin Proteins in Lupin-Enriched Pasta by Untargeted and Targeted Mass Spectrometry

Foods ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1083
Author(s):  
Gilda Aiello ◽  
Yuchen Li ◽  
Giovanna Boschin ◽  
Marco Stanziale ◽  
Carmen Lammi ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Multiple Reaction Monitoring ◽  
Protein Profile ◽  
Tandem Mass ◽  
Label Free ◽  
Narrow Leaf ◽  
Ionization Tandem Mass Spectrometry

The supplementation of different food items with grain legumes and, in particular, with lupin has been demonstrated to provide useful health benefits, especially in the area of cardiovascular disease prevention. In this work, label free quantitative untargeted and targeted approaches based on liquid chromatography−electrospray ionization−tandem mass spectrometry (LC−ESI−MS/MS) for investigating the protein profile of three pasta samples containing different percentages of narrow-leaf lupin flour were carried out. The untargeted method permitted the identification of the main acidic globulins (α-conglutin, β-conglutin, and δ-conglutin) and the comparison of their profile with raw lupin flour. The targeted method, based on High-performance liquid chromatography electrospray ionization tandem mass spectrometry HPLC-Chip-Multiple Reaction Monitoring (MRM) mode, allowed the quantification of γ-conglutin, the main hypoglycemic component of lupin protein: its concentration was around 2.25 mg/g in sample A, 2.16 mg/g in sample D, and 0.57 mg/g in sample F.

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