Plasma N-acetylaspartate: Development and validation of a quantitative assay based on HPLC-MS-MS and sample derivatization
| Autor: | Beatrice Campi, Antonella Marvelli, Giovanni Ceccarini, Simone Codini, Riccardo Zucchi, Alessandro Saba, Giuseppe Daniele, Ele Ferrannini, Ferruccio Santini |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Sample (material) Clinical Biochemistry Urine Tandem mass spectrometry Biochemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cerebrospinal fluid Tandem Mass Spectrometry Humans Sample preparation Derivatization Chromatography High Pressure Liquid Aspartic Acid Reproducibility Chromatography Chemistry Biochemistry (medical) Reproducibility of Results General Medicine Plasma 030104 developmental biology 030220 oncology & carcinogenesis Chromatography Liquid |
| Zdroj: | Clinica Chimica Acta. 508:146-153 |
| ISSN: | 0009-8981 |
| DOI: | 10.1016/j.cca.2020.05.020 |
| Popis: | N-acetylaspartate is a human endogenous compound synthesized by neurons, which is involved in neuronal metabolism. It is used as a marker in brain magnetic resonance spectroscopy to investigate several neurological and metabolic disorders, that can be related to a variation of its concentration with respect to reference values. N-acetylaspartate is present also in biological fluids, such as plasma, urine, and cerebrospinal fluid, where it can be quantified. Here we describe the development and validation, in compliance with the EMA guidelines, of a novel assay method for the quantification of N-acetylaspartate in plasma based on tandem mass spectrometry coupled to liquid chromatography. Its peculiarity lies in the fact that sample preparation includes an esterification step, which significantly improves the chromatographic performances and, consequently, also the method sensitivity, reproducibility and accuracy. Instrumental LLOQ is 0.06 ng/mL, i.e. at least 300 times lower than the medium N-acetylaspartate concentration in samples, accuracy is in the range 98–103%, while precision lies between 1 and 3%. The method robustness was tested in about 1000 injections of plasma samples, 96 of which were used also to assess the reference ranges in control subjects (16.46–63.40 ng/mL). Controls were then compared to plasma samples from type 2 diabetic patients. Contrary to brain magnetic resonance spectroscopy, which demonstrated a decrease in the N-acetylaspartate levels in right frontal and parieto-temporal region of type 2 diabetic patients, plasma analysis showed no statistical difference with respect to controls. However, the method here described can be profitably used in studies concerning different disorders with CNS involvement, as confirmed by reports available in the literature. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect