Quantification of glyoxal, methylglyoxal and 3-deoxyglucosone in blood and plasma by ultra performance liquid chromatography tandem mass spectrometry: evaluation of blood specimen.

Author(s): Scheijen JL, Schalkwijk CG

Publication: Clin Chem Lab Med, 2014, Vol. 52, Page 85-91

PubMed ID: 23492564 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of sodium fluoride (NaF), perchloric acid (PCA) precipitation of proteins, room temperature storage duration, and assaying plasma rather than whole blood on levels of glyoxal, methylglyoxal and 3-deoxyglucosone in specimens from patients with or without type II diabetes.

Conclusion of Paper

When blood was not precipitated with PCA, longer room temperature storage of specimens anticoagulated with both EDTA and NaF led to increasing glyoxal and methylglyoxal levels and decreasing 3-deoxyglucosone levels in blood, and for specimens anticoagulated with EDTA alone, methylglyoxal increased and 3-deoxyglucosone levels decreased with longer room temperature storage of blood. In contrast, when the blood was precipitated with PCA, room temperature storage of specimens had no effects on any of the three analytes when measured in whole blood. With increasing storage of EDTA blood at room temperature, plasma levels of glyoxal and 3-deoxyglucosone decreased, but methylglyoxal levels in plasma remained stable. When blood that was anticoagulated with EDTA and NaF was stored at room temperature, plasma levels of methylglyoxal increased and 3-deoxyglucosone decreased with longer storage durations, but glyoxal levels in plasma remained stable. When EDTA plasma was stored at room temperature, specimens not immediately precipitated with PCA had decreased levels of glyoxal and 3-deoxyglucosone after 60 min, but when the plasma was immediately precipitated with PCA, 120 min of room temperature storage had no effect on plasma levels of glyoxal, methylglyoxal or 3-deoxyglucosone. Levels of methylglyoxal and 3-deoxyglucosone were higher in plasma from patients with type-II diabetes than patients without diabetes.

Studies

Study Purpose

The purpose of this study was to determine the effects of NaF addition, PCA precipitation of proteins, room temperature storage duration, and assaying plasma rather than whole blood on levels of glyoxal, methylglyoxal and 3-deoxyglucosone in specimens from patients with or without type II diabetes. Some plasma specimens were frozen at an unspecified temperature before analysis.

Summary of Findings:

When blood was not precipitated with PCA, longer room temperature storage of specimens anticoagulated with both EDTA and NaF led to increasing glyoxal and methylglyoxal levels and decreasing 3-deoxyglucosone levels in blood, and for specimens anticoagulated with EDTA alone, methylglyoxal increased and 3-deoxyglucosone levels decreased with longer room temperature storage of blood. In contrast, when the blood was precipitated with PCA, room temperature storage of whole blood had no effect on any of the three analytes. With longer storage of EDTA blood at room temperature, plasma levels of glyoxal and 3-deoxyglucosone decreased, but methylglyoxal levels in plasma remained stable. When blood that was anticoagulated with EDTA and NaF was stored at room temperature, plasma levels of methylglyoxal increased and 3-deoxyglucosone decreased with longer storage durations, but glyoxal levels in plasma remained stable. When EDTA plasma was stored at room temperature, specimens not immediately precipitated with PCA had decreased levels of glyoxal and 3-deoxyglucosone after 60 min, but when the plasma was immediately precipitated with PCA, 120 min of room temperature storage had no effect on plasma levels of glyoxal, methylglyoxal or 3-deoxyglucosone. Levels of methylglyoxal and 3-deoxyglucosone were higher in plasma from patients with type-II diabetes than patients without diabetes.

Biospecimens

Preservative Types

None (Fresh)
Frozen

Diagnoses:

Diabetes Type 2
Not specified

Platform:

Analyte	Technology Platform
Carbohydrate	LC-MS or LC-MS/MS

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Aliquots and Components	Blood and blood products	Plasma Whole blood
Biospecimen Acquisition	Anticoagulant	EDTA Sodium fluoride/EDTA
Storage	Time at room temperature	0 min 30 min 60 min 90 min 120 min
Analyte Extraction and Purification	Analyte isolation method	PCA precipitation No precipitation

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