Identification and quantification of glucose degradation products in heat-sterilized glucose solutions for parenteral use by thin-layer chromatography

Sarah Leitzen,Thomas Zapf,Martin Brandl,Matthias Vogel,Anette Engels,A M Abd El-Aty

doi:10.1371/journal.pone.0253811

Abstract

During heat sterilization of glucose solutions, a variety of glucose degradation products (GDPs) may be formed. GDPs can cause cytotoxic effects after parenteral administration of these solutions. The aim of the current study therefore was to develop a simple and quick high-performance thin-layer chromatography (HPTLC) method by which the major GDPs can be identified and (summarily) quantified in glucose solutions for parenteral administration. All GDPs were derivatized with o-phenylenediamine (OPD). The resulting GDP derivatives (quinoxalines) were applied to an HPTLC plate. After 20 minutes of chamber saturation with the solvent, the HPTLC plate was developed in a mixture of 1,4-dioxane-toluene-glacial acetic acid (49:49:2, v/v/v), treated with thymol-sulfuric acid spray reagent, and heated at 130°C for 10 minutes. Finally, the GDPs were quantified by using a TLC scanner. For validation, the identities of the quinoxaline derivatives were confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Glyoxal (GO)/methylglyoxal (MGO) and 3-deoxyglucosone (3-DG)/3-deoxygalactosone (3-DGal) could be identified and quantified in pairs, glucosone (2-KDG), 5-hydroxymethylfurfural (5-HMF), and 3,4-dideoxyglucosone-3-ene (3,4-DGE) each individually. For 2-KDG, the linearity of the method was demonstrated in the range of 1–50 μg/mL, for 5-HMF and 3,4-DGE 1–75 μg/mL, for GO/MGO 2–150 μg/mL, and for 3-DG/3-DGal 10–150 μg/mL. All GDPs achieved a limit of detection (LOD) of 2 μg/mL or less and a limit of quantification (LOQ) of 10 μg/mL or less. R2 was 0.982 for 3.4-DGE, 0.997 for 5-HMF, and 0.999 for 2-KDG, 3-DG/3-DGal, and GO/MGO. The intraday precision was between 0.4 and 14.2% and the accuracy, reported as % recovery, between 86.4 and 112.7%. The proposed HPTLC method appears to be an inexpensive, fast, and sufficiently sensitive approach for routine quantitative analysis of GDPs in heat-sterilized glucose solutions.

Highlights

Sterile glucose solutions are commonly used as reconstitution solvents or diluents for injectable drugs and for peritoneal dialysis solutions [1]
Complete separation of glucose degradation products (GDPs) upon prior derivatization with OPD has been described for various (U)HPLC methods [10, 24, 27, 28]
The derivatization reagent OPD was chosen for this high-performance thin-layer chromatography (HPTLC) method, too

Summary

Introduction

Sterile glucose solutions are commonly used as reconstitution solvents or diluents for injectable drugs and for peritoneal dialysis solutions [1]. Indications for the use of glucose solutions range from the treatment of carbohydrate deficiencies and fluid losses, carrier solutions for compatible drugs and electrolyte concentrates, additions of free water, energy supplies, hypoglycemic states, and high-calorie intake when fluids are restricted to carbohydrate components in parenteral nutrition [2]. For the standard autoclaving process at 121 ̊C, a sterility assurance level of 10−6 is typically achieved; at the same time glucose degradation products (GDPs) may be formed during such heat treatment, negatively affecting the quality of glucose solutions [7,8,9]. Most GDPs can be deduced from glucose after oxidative and dehydration processes caused by the effect of heat

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Conclusion