Simultaneous Determination of 5'-Monophosphate Nucleotides in Infant Formulas by HPLC-MS

• Published in: Journal of chromatographic science Vol. 49; no. 4; pp. 332 - 337
• Main Authors: Ren, Yiping, Chen, Xiaochun, Zhang, Jingshun, Li, Duo, Song, Xiaodan
• Format: Journal Article
• Language: English
• Published: Niles, IL Oxford University Press 01-04-2011; Preston Publications
• Subjects: Acetonitriles; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Food industries; Fundamental and applied biological sciences. Psychology; Humans; Infant; Infant Formula - chemistry; Linear Models; Purine Nucleotides - analysis; Pyrimidine Nucleotides - analysis; Reproducibility of Results; Research and development. New food products, dietetic foods and beverages; Sensitivity and Specificity; Tandem Mass Spectrometry - methods; Purine nucleotide; Tandem mass spectrometry; Chemical analysis; AMP; Coupled method; HPLC chromatography; GMP; Electrospray; Reversed phase chromatography; Simultaneous measurement; Infant formula; Pyrimidine nucleotide; Ribonucleotide; Mass spectrometry; Quantitative analysis
• ISSN: 0021-9665; 1945-239X
• DOI: 10.1093/chrsci/49.4.332

A method was developed for simultaneous determination of 5'-monophosphate nucleotides, adenosine 5'-monophosphate, cytidine 5'-monophosphate, guanosine 5'-monophosphate, inosine 5'-monophosphate, and uridine 5'-monophosphate in infant formulas by high-performance liquid chromatography-mass spectrometry equipped with electrospray ionization source. The complete chromatographic separation of five nucleotides was achieved through a Symmetry C18 column, after a binary gradient elution with water containing 0.1 % formic acid and acetonitrile as mobile phase. The multi-reaction monitoring mode was applied for tandem mass spectrometry analysis. The established method was further validated by determining the linearity (R2 > 0.999), recovery (92.0-105.0%), and precision (relative standard deviation ≤ 6.97%). To verify the applicability of the method, thirty commercially available infant formulas were randomly purchased from the supermarkets in Hangzhou, China, and then analyzed. The results showed that the developed method is validated, sensitive, and reliable for quantitation of nucleotides in infant formulas.