Traceable Amino Acid Analyses of Proteins and Peptides by Isotope-Dilution Mass Spectrometry Using Precolumn Derivatization Reagent

Kinumi, Tomoya; Ichikawa, Ryota; Arimoto, Hirokazu; Takatsu, Akiko

doi:10.2116/analsci.26.1007

Cited by 20 publications

(16 citation statements)

References 8 publications

(12 reference statements)

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“…The measurement was conducted under a selected reaction monitoring (SRM) mode using the following 14. Other conditions were described previously [12]. For HILIC/MS analysis, 2 μL of sample solution was injected into an LC/MS system, LC/MS-2010-EV (Shimadzu, Kyoto, Japan) using a ZIC-HILIC column (2.1 mm ID × 250 mm, Merk Chemicals, Darmstadt, Germany) and equipped with an electrospray ion source.…”

Section: Hydrolysis and Amino Acid Analyses By Pre-column Lc-ms/ Ms Amentioning

confidence: 99%

“…For this analysis, hydrophilic chromatography-mass spectrometry (HILIC-MS) was utilized [11]. We have also developed ID MS-based amino acid analysis methods, including precolumn derivatization liquid chromatography-tandem mass spectrometry (LC-MS/MS) [12]. The advantage of this method is that it allows us to quantify hydrophilic amino acids such as glutamic acid and aspartic acid, with a rapid and precise measurement.…”

Section: Introductionmentioning

confidence: 99%

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Development of SI-traceable C-peptide certified reference material NMIJ CRM 6901-a using isotope-dilution mass spectrometry-based amino acid analyses

et al. 2012

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A certified reference material (CRM) is a higher-order calibration material used to enable a traceable analysis. This paper describes the development of a C-peptide CRM (NMIJ CRM 6901-a) by the National Metrology Institute of Japan using two independent methods for amino acid analysis based on isotope-dilution mass spectrometry. C-peptide is a 31-mer peptide that is utilized for the evaluation of β-cell function in the pancreas in clinical testing. This CRM is a lyophilized synthetic peptide having the human C-peptide sequence, and contains deamidated and pyroglutamylated forms of C-peptide. By adding water (1.00 ± 0.01) g into the vial containing the CRM, the C-peptide solution in 10 mM phosphate buffer saline (pH 6.6) is reconstituted. We assigned two certified values that represent the concentrations of total C-peptide (mixture of C-peptide, deamidated C-peptide, and pyroglutamylated C-peptide) and C-peptide. The certified concentration of total C-peptide was determined by two amino acid analyses using pre-column derivatization liquid chromatography-mass spectrometry and hydrophilic chromatography-mass spectrometry following acid hydrolysis. The certified concentration of C-peptide was determined by multiplying the concentration of total C-peptide by the ratio of the relative area of C-peptide to that of the total C-peptide measured by liquid chromatography. The certified value of C-peptide (80.7 ± 5.0) mg/L represents the concentration of the specific entity of C-peptide; on the other hand, the certified value of total C-peptide, (81.7 ± 5.1) mg/L can be used for analyses that does not differentiate deamidated and pyroglutamylated C-peptide from C-peptide itself, such as amino acid analyses and immunochemical assays.

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Section: Hydrolysis and Amino Acid Analyses By Pre-column Lc-ms/ Ms Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of SI-traceable C-peptide certified reference material NMIJ CRM 6901-a using isotope-dilution mass spectrometry-based amino acid analyses

et al. 2012

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“…3.2). Of among, complete hydrolysis is critical for overall accuracy of the procedure (Albin et al, 2000;Anderson et al, 1977;Darragh et al, 1996;Fountoulakis and Lahm, 1998;Kinumi et al, 2010). Moreover, for the analysis of an unknown protein, optimization of hydrolysis conditions should be considered before conducting the whole procedure.…”

Section: Amino Acid Analysismentioning

confidence: 99%

Development of Metrology for Modern Biology

Park¹,

Choi²,

Jeong³

2012

Modern Metrology Concerns

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“…To ensure the reliability of amino acid analyses, the National Metrology Institute of Japan at the National Institute of Advanced Industrial Science and Technology (NMIJ/AIST) has developed high-purity certified reference materials (CRMs) for 17 proteinogenic amino acids 1 relevant to clinical chemistry, 2,3 food chemistry, [4][5][6] pharmaceutical biotechnology, 7 and the accurate quantification of protein/peptides [8][9][10] by amino acid analysis. These CRMs will be the primary standards used in the metrological traceability system, and exhibit purities of greater than 0.997 kg kg -1 , with uncertainties of less than 0.003 kg kg -1 .…”

Section: Introductionmentioning

confidence: 99%

Effects of the pH and Concentration on the Stability of Standard Solutions of Proteinogenic Amino Acid Mixtures

et al. 2017

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To prepare metrologically traceable amino acid mixed standard solutions, it is necessary to determine the stability of each amino acid present in the mixed solutions. In the present study, we prepared amino acid mixed solutions using certified reference standards of 17 proteinogenic amino acids, and examined the stability of each of these amino acids in 0.1 N HCl. We found that the concentration of glutamic acid decreased significantly during storage. LC/MS analysis indicated that the instability of glutamic acid was due to the partial degradation of glutamic acid to pyroglutamic acid in 0.1 N HCl. Using accelerated degradation tests, we investigated several solvent compositions to improve the stability of glutamic acid in amino acid mixed solution, and determined that the change of the pH by diluting the mixed solution improved the stability of glutamic acid.

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Traceable Amino Acid Analyses of Proteins and Peptides by Isotope-Dilution Mass Spectrometry Using Precolumn Derivatization Reagent

Cited by 20 publications

References 8 publications

Development of SI-traceable C-peptide certified reference material NMIJ CRM 6901-a using isotope-dilution mass spectrometry-based amino acid analyses

Development of SI-traceable C-peptide certified reference material NMIJ CRM 6901-a using isotope-dilution mass spectrometry-based amino acid analyses

Development of Metrology for Modern Biology

Effects of the pH and Concentration on the Stability of Standard Solutions of Proteinogenic Amino Acid Mixtures

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