Quantification of rice α‐globulin allergen using liquid chromatography–tandem mass spectrometry combined with cysteine‐specific modifier and extended stable isotope‐labeled peptide

2022 ◽  
Author(s):  
Huan Yang ◽  
Zhao‐yun Cao ◽  
Ren‐xiang Mou ◽  
Zhen‐zhen Cao ◽  
Ming‐xue Chen
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

scholarly journals Measurement of Urinary d- and l-2-Hydroxyglutarate Enantiomers by Stable-Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry after Derivatization with Diacetyl-l-Tartaric Anhydride

2004 ◽  
Vol 50 (8) ◽  
pp. 1391-1395 ◽  
Author(s):  
Eduard A Struys ◽  
Erwin E W Jansen ◽  
Nanda M Verhoeven ◽  
Cornelis Jakobs
Keyword(s):  
Mass Spectrometry ◽  
Differential Diagnosis ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Internal Standards ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Abstract Background: The differential diagnosis of d-2-hydroxyglutaric aciduria (d-2-HGA), l-2-hydroxyglutaric aciduria (l-2-HGA), and the combined d/l-2-hydroxyglutaric aciduria (d/l-2-HGA) can be accomplished only by the measurement of the corresponding 2-hydroxyglutarate (2-HG). Available methods for the determination of d- and l-2-HG in urine are either time-consuming and expensive or have not been extensively validated. We aimed to develop a method for their rapid and sensitive measurement. Methods: We used liquid chromatography–tandem mass spectrometry (LC-MS/MS) for the determination of d- and l-2-HG with stable-isotope-labeled internal standards. Urine samples of 20 μL were mixed with 250 μL of methanol containing the internal standards and subsequently dried under nitrogen. The analytes were derivatized by use of diacetyl-l-tartaric anhydride (DATAN) to obtain diastereomers, which were separated on an achiral C18 HPLC column and detected by MS/MS in multiple-reaction-monitoring mode. Results: The use of DATAN as chiral derivatization reagent provided very well separated peaks of the formed diastereomers of d- and l-2-HG, with a total runtime of 5 min. The inter- and intraassay CVs for d- and l-2-HG ranged from 3.4% to 6.2%. Mean recoveries of d- and l-2-HG, evaluated on two concentrations, were 94%. Detection limit of the presented method was 20 pmol for a sample volume of 20 μL. Method comparison of the LC-MS/MS method with a gas chromatography–mass spectrometry method, in which d- and l-2-HG were derivatized with R-(−)-butanol, showed good agreement between the two methods. Conclusions: Urinary d- and l-2-HG can be analyzed by MS/MS after derivatization with DATAN. The presented method may be suitable for the differential diagnosis of 2-HGA.

scholarly journals Feasibility of Utilizing Stable-Isotope Dimethyl Labeling in Liquid Chromatography–Tandem Mass Spectrometry-Based Determination for Food Allergens—Case of Kiwifruit

Molecules ◽  
2019 ◽  
Vol 24 (10) ◽  
pp. 1920 ◽  
Author(s):  
Yi-Chen Shih ◽  
Jhih-Ting Hsiao ◽  
Fuu Sheu
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Food Allergens ◽  
Chromatography Tandem Mass Spectrometry ◽  
Dimethyl Labeling ◽  
Liquid Chromatography Tandem Mass ◽  
Signature Peptides

Stable-isotope dimethyl labeling is a highly reactive and cost-effective derivatization procedure that could be utilized in proteomics analysis. In this study, a liquid chromatography– tandem mass spectrometry in multiple reaction monitoring mode (LC-MS-MRM) platform for the quantification of kiwi allergens was first developed using this strategy. Three signature peptides for target allergens Act d 1, Act d 5, and Act d 11 were determined and were derivatized with normal and deuterated formaldehyde as external calibrants and internal standards, respectively. The results showed that sample preparation with the phenol method provided comprehensive protein populations. Recoveries at four different levels ranging from 72.5–109.3% were achieved for the H-labeled signature peptides of Act d 1 (SPA1-H) and Act d 5 (SPA5-H) with precision ranging from 1.86–9.92%. The limit of quantification (LOQ) was set at 8 pg mL−1 for SPA1-H and at 8 ng mL−1 for SPA5-H. The developed procedure was utilized to analyze seven kinds of hand-made kiwi foods containing 0.0175–0.0515 mg g−1 of Act d 1 and 0.0252–0.0556 mg g−1 of Act d 5. This study extended the applicability of stable-isotope dimethyl labeling to the economical and precise determination of food allergens and peptides.

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