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Validation of an HPLC method for the determination of nucleotides in infant formulae (CROSBI ID 153578)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Krpan, Marina ; Vahčić, Nada ; Hruškar, Mirjana Validation of an HPLC method for the determination of nucleotides in infant formulae // Journal of liquid chromatography & related technologies, 32 (2009), 2747-2755

Podaci o odgovornosti

Krpan, Marina ; Vahčić, Nada ; Hruškar, Mirjana

engleski

Validation of an HPLC method for the determination of nucleotides in infant formulae

This paper describes the development and validation of simple, fast and sensitive high performance liquid chromatographic method for the determination of the 5’ -mononucleotides ; adenosine 5’ -monophosphate, cytidine 5’ -monophosphate, guanosine 5’ -monophosphate, inosine 5’ -monophosphate and uridine 5’ -monophosphate in infant formulae in defined labour conditions. Following deproteinisation and filtration, the sample extract was analysed by reversed-phase liquid chromatography. The method was developed by using reverse-phase column C18. Isocratic elution was used with mobile phase consisted of 0.1 M potassium phosphate buffer and 4 mM tetrabutylammonium hydrogen phosphate. The pH of solution was adjusted to 6.0. The analysis of nucleotides was performed on a PDA detector at 260 and 278 nm. Analytical validation parameters such as specificity and selectivity, linearity, accuracy, precision, robustness and system suitability were evaluated. During evaluation of the analytical parameters, it was shown that the method is linear (r = 0, 999). The recoveries ranged 100 ± 3 and the relative standard deviation was ≤ 3. The precision of the method was achieved with a coefficient of variation (CV %), which is less than 3 %. Standard solutions are stable during 30 hours and in range of method robustness. Validation of HPLC method for determination of nucleotides has showed that developed analytical method is acceptable for its intended purpose in defined labour conditions.

HPLC; infant formulae; method validation; nucleotides

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Podaci o izdanju

32

2009.

2747-2755

objavljeno

1082-6076

Povezanost rada

Prehrambena tehnologija

Indeksiranost