Β-hydroxymyristic acid as a chemical marker to detect endotoxins in dialysis water.
Anal Biochem
; 470: 71-7, 2015 Feb 01.
Article
en En
| MEDLINE
| ID: mdl-25449302
ABSTRACT
An analytical chemical method has been developed for determination of ß-hydroxymyristic acid (ß-HMA), a component of lipopolysaccharides (LPSs/endotoxins) in dialysis water. In our investigation, the ß-HMA component was used as a chemical marker for endotoxin presence in dialysis water because it is available in the molecular subunit (lipid A) and responsible for toxicity. It is the most abundant saturated fatty acid in that subunit. The developed method is based on fluorescence derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ). A high-performance liquid chromatographic separation of the ß-HMA derivative was achieved using an octadecyl silica column in gradient elution. A wide dynamic range of ß-HMA was tested and a calibration curve was constructed with accuracy of 90% and variability of less than 10%. The limits of detection and quantification obtained were 2 and 5µM, respectively. The developed method was applied to detect endotoxins in dialysis water by alkaline hydrolysis of LPS using NaOH (0.25M) at 60°C for 2h. After hydrolysis, free acid was detected as its NBD-PZ derivative using high-performance liquid chromatography/mass spectrometry (HPLC/MS). Good recovery rates ranging from 98 to 105% were obtained for ß-HMA in dialysis water.
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Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Agua
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Técnicas de Química Analítica
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Lipopolisacáridos
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Diálisis Renal
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Ácidos Mirísticos
Idioma:
En
Revista:
Anal Biochem
Año:
2015
Tipo del documento:
Article
País de afiliación:
Francia