Lipidperoxidation In Parenteral Nutrition: Sensitivity of a Novel Potential Routine Lab Method And Its Compliance To Efsa Requirements

Introduction: The extent of lipid peroxidation (LPO) is critical for the safe use of in all-in-one (AiO) parenteral nutrition (PN) admixtures. Admixed nutrients, such as trace elements or drugs, can further induce LPO. However, no well-defined and rapid lab assay exists for routine quantification of the most relevant toxic end products. Polyunsaturated fatty acids (PUFA), essential lipid components in PN and regular food, are highly susceptible to (per-)oxidation, forming toxic byproducts like malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE), and 4-hydroxy-2-nonenal (HNE). These compounds compromising PN quality and pose potential health risks, particularly for vulnerable PN-dependent patients.

Objectives: The aim is to develop and characterize a novel assay capable of simultaneously detect and quantify these three key byproducts with high stability, reliability, and sensitivity.

Methods: HPLC-UV quantification with a Luna C8(2) 50mm x 3 mm, 3µm particles column in a gradient mode (H₂O: AcN 95% to 5%) and 3D-field UV-VIS spectra for qualitative analysis. Pure standards and Smofkabiven®​-EF (AiO PN) were used. Derivatizing chemical: 3-methyl-2-benzothiazo-linone-hydrazone (MBTH).

Results: The target substances were well separated and identified by the different retention times (MDA-MBTH 5.33; HHE-MBTH 6.01; HNE- MBTH 6.55). Total sample preparation time was 12 min, with a short 5 min incubation yelding stable derivatives, allowing combined sample analyses. The method was rapid, demonstrated consistent retention times (RSD<0.04%), a robust correlation between area and injected amount of probes, high sensitivity (detection limit 2 µg/ml from a 100µl PN sample, with potential for further lowering), high specificity, with no apparent spectral or chromatographic interferences, and compliant to the EFSA Threshold of Toxicological Concern (TTC) range of 1.5, 30, and 1.5 µg/kg/d for oral MDA, HHE, and HNE, respectively.

Conclusion: The novel, sensitive LC-UV method provides precise, reliable, and simultaneous quantification of MDA, HHE, HNE, offering a robust framework for analyzing LPO formation in lipid-containing nutritional products. The method assesses relevant safety standards, and has the potential to monitor LPO levels in AiO PN admixtures also in presence of additives like critical nano-iron drugs, which may require co-administration due to limited venous access. This method will be a prerequisite for the necessary stability and compatibility assessment.

Interessenkonflikte

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Publication History

Article published online:
25 May 2025