Identification and Quantification of Glucose Degradation Products in Heat-Sterilized Glucose Solutions for Parenteral Use

Moist heat sterilization of glucose solutions inevitably leads to the formation of various glucose degradation products (GDPs). Upon parenteral administration, these GDPs may have toxic effects due to their high reactivity. The aims of the current PhD project therefore were:


(I) to investigate whether alternative time-temperature settings (of equivalent sterilization efficacy) during moist heat sterilization affect the formation of GDPs in parenteral glucose solutions as detected by an adapted and validated high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach


(II) to compare GDP concentrations of selected commercially available 5-50 % glucose solutions for parenteral use with those observed in (I) and, if appropriate, propose an autoclaving protocol for glucose solutions


(III) to compile literature toxicity data for the observed GDPs in order to evaluate the GDP patterns that changed during autoclaving with alternative settings in terms of overall toxic potential


(IV) 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


Trace amounts of the GDPs glyoxal (GO), methylglyoxal (MGO), glucosone (2-KDG), 3-deoxyglucosone/3-deoxygalactosone (3-DG/3-DGal), 3,4-dideoxyglucosone-3-ene (3,4-DGE), and 5-hydroxymethylfurfural (5-HMF) could be quantified after derivatization with ophenylenediamine (OPD) by using the LC-MS/MS method. While autoclaving at 121 °C for 18 min resulted in the lowest levels of 3-DG/3-DGal and 5-HMF, autoclaving at 116 °C for 57 min yielded the lowest levels of GO and MGO.


Industrially manufactured glucose solutions showed an up to 10-fold higher content of GO, whereas the GDP levels of MGO, 2-KDG, 3-DG/3-DGal, 3,4-DGE, and 5-HMF were significantly lower in industrial products than in the self-prepared solutions. By employing the new, validated HPTLC method, GO/MGO and 3-DG/3-DGal could be quantified in pairs and 2-KDG, 3,4-DGE, and 5-HMF each individually with good sensitivity.


In conclusion, the levels of GDPs formed during heat sterilization of glucose solutions should be reduced, regulated, and monitored. According to the German national monographs (German standard marketing authorizations), there is a limit value only for 5- HMF, which was met in all marketed glucose solutions examined. The measured contents of the most toxic GDPs GO and MGO, however, exceeded the scientific recommendations of WHO, ECHA, and BfR. In order to reduce GO levels, we recommend the use of an alternative autoclave protocol of equivalent sterility assurance (e.g., 116 °C and F0 = 57 min). In addition, limits should be established for all GDPs associated with health concerns. The HPTLC method developed and validated in this work appears to be a suitable tool to test for compliance with such limits.