Development and Application of an HILIC-MS/MS Method for the Quantitation of Nucleotides in Infant Formula

Inoue, Kōichi; Obara, Rutsuko; Hino, Tomoaki; Oka, Hisao

doi:10.1021/jf102023p

Cited by 52 publications

(42 citation statements)

References 21 publications

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“…They have been widely used in the food industry . They can enhance immunity and decrease the risk of diarrhea and sepsis . The addition of 5′‐ribonucleotides in infant formula can improve the composition of intestinal microflora, emulating the attibute of human milk …”

Section: Introductionmentioning

confidence: 99%

Combined ion exchange and adsorption equilibria of 5′-ribonucleotides on the strong acid cation exchange resin NH-1

Jiao

Wang

et al. 2017

J. Chem. Technol. Biotechnol

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BACKGROUND 5′‐ribonucleotides including uridine 5′‐monophosphate (UMP), adenosine 5′‐monophosphate (AMP), cytidine 5′‐monophosphate (CMP), and guanosine 5′‐monophosphate (GMP) have been widely used as food additives. Ion exchange is a potential technique to separate 5′‐ribonucleotides. A knowledge of uptake equilibrium on ion exchangers is important for the design and scale‐up of ion exchange processes. The uptake equilibria of 5′‐ribonucleotides and Na+ on the strong acid cation exchange resin NH‐1 were studied experimentally and modelled in binary and multicomponent systems. The uptake mechanism and thermodynamics were also studied. RESULTS A modified Helfferich model was developed combining ion exchange of cations with adsorption of zwitter ions. The model can predict the uptake equilibria of 5′‐ribonucleotides and Na+ successfully in multicomponent systems. The hydrophobicity of bases of 5′‐ribonucleotides determined their relative affinities to the resin. The resin shows the strongest affinity for AMP among 5′‐ribonucleotides. The uptakes of 5′‐ribonucleotides cations is an endothermic and entropy‐driven process. CONCLUSION The equilibrium model proposed is important for simulation and design of the dynamic separation process of 5′‐ribonucleotides. UMP, GMP, and CMP can be separated based on their difference in isoelectric point. AMP and CMP with similar isoelectric point can be separated based on their difference in affinity to the resin. © 2016 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Combined ion exchange and adsorption equilibria of 5′-ribonucleotides on the strong acid cation exchange resin NH-1

Jiao

Wang

et al. 2017

J. Chem. Technol. Biotechnol

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“…IPRs that interact with nucleobase amino groups such as n-alkyl perfluorinated carboxylic acids have also been used for LC-MS separation (Feng et al , 2008). Other alternative LC-MS techniques are based on mixed-mode reversed-phase/weak anion exchange (Hinterwirth et al , 2010), hydrophilic interaction chromatography (Inoue et al , 2010) or affinity chromatography (Kammerer et al , 2005a; Kammerer et al , 2005b). In addition to LC separations, capillary electrophoresis coupled to MS has also been used for detection of nucleotides (Soo et al , 2004)…”

Section: Introductionmentioning

confidence: 99%

High information throughput analysis of nucleotides and their isotopically enriched isotopologues by direct-infusion FTICR-MS

et al. 2011

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Fourier transform-ion cyclotron resonance-mass spectrometry (FTICR-MS) is capable of acquiring unmatched quality of isotopologue data for stable isotope resolved metabolomics (SIRM). This capability drives the need for a continuous ion introduction for obtaining optimal isotope ratios. Here we report the simultaneous analysis of mono and dinucleotides from crude polar extracts by FTICR-MS by adapting an ion-pairing sample preparation method for LC-MS analysis. This involves a rapid cleanup of extracted nucleotides on pipet tips containing a C18 stationary phase, which enabled global analysis of nucleotides and their 13C isotopologues at nanomolar concentrations by direct infusion nanoelectrospray FTICR-MS with 5 minutes of data acquisition. The resolution and mass accuracy enabled computer-assisted unambiguous assignment of most nucleotide species, including all phosphorylated forms of the adenine, guanine, uracil and cytosine nucleotides, NAD+, NADH, NADP+, NADPH, cyclic nucleotides, several UDP-hexoses, and all their 13C isotopologues. The method was applied to a SIRM study on human lung adenocarcinoma A549 cells grown in [U-13C] glucose with or without the anti-cancer agent methylseleninic acid. At m/z resolving power of 400,000, 13C-isotopologues of nucleotides were fully resolved from all other elemental isotopologues, thus allowing their 13C fractional enrichment to be accurately determined. The method achieves both high sample and high information throughput analysis of nucleotides for metabolic pathway reconstruction in SIRM investigations.

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“…The total nucleotide level ranged from nd to 600.2 µg/g in powder infant formula. For additional information regarding the study, see Inoue et al (2).…”

Section: Resultsmentioning

confidence: 99%

HILIC-MS/MS Method for the Quantitation of Nucleotides in Infant Formula and Adult Nutritional Formula: First Action 2011.21

Inoue

Dowell²

2012

Journal of AOAC INTERNATIONAL

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Official Method 2011.21 is for the quantitation of the following nucleotides: adenosine 5'-monophosphate (AMP), guanosine 5'-monophosphate (GMP), uridine 5'-monophosphate (UMP), cytidine 5'-monophosphate (CMP), and inosine 5'-monophosphate (IMP) in infant formula and adult/pediatric nutritional formula. It uses hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS). Preparation of the internal standards was conducted using centrifugal ultrafiltration and the standards are AMP- (13)C10, (15)N5; GMP-(13)C10, (15)N5; UMP-(13)C9, (15)N2; and15 CMP- (13)C9, (15)N3. Data were collected by using multiple reaction monitoring of the product ions of protonated molecules of the five nucleotides generated by positive-electrospray ionization. The HILIC conditions were conducted with ammonium formate (30 mmol/L) in water (pH 2.5, adjusted with formic acid) and methanol. The LOD and LOQ of the standard solution were 0.005-0.01 and 0.01-0.03 microg/mL, respectively. Recovery data were collected for intraday and interday testing and ranged from 98.1 to 108.9% with an RSD of 0.7-5.4%. The analytical range of the method is between 0.04 to 5 microg/mL for standard solution.

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Development and Application of an HILIC-MS/MS Method for the Quantitation of Nucleotides in Infant Formula

Cited by 52 publications

References 21 publications

Combined ion exchange and adsorption equilibria of 5′-ribonucleotides on the strong acid cation exchange resin NH-1

Combined ion exchange and adsorption equilibria of 5′-ribonucleotides on the strong acid cation exchange resin NH-1

High information throughput analysis of nucleotides and their isotopically enriched isotopologues by direct-infusion FTICR-MS

HILIC-MS/MS Method for the Quantitation of Nucleotides in Infant Formula and Adult Nutritional Formula: First Action 2011.21

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