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

Autori
Krpan, Marina ; Vahčić, Nada ; Hruškar, Mirjana

Izvornik
Journal of liquid chromatography & related technologies (1082-6076) 32 (2009); 2747-2755

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
HPLC; infant formulae; method validation; nucleotides

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Prehrambena tehnologija

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