Award
Dr Larissa Balakireva, CEO & Founder of NovoCIB, was awarded with the Trophy of
"Femmes en Or 2011, Femme de l'Innovation"
in September 2011
.

Nucleotide ion-pair reverse-phase HPLCYeast Nucleotide HPLC Analysis

 
Ref.
Price per sample*
Yeast Free Bases, Nucleosides, Nucleotides and RNA & Bases
#S1100-03
€ 450.00

* Pricing updated September 08th, 2015
To order Nucleotide Analysis by HPLC click here or ask for Quotation

Yeast Extracts Enriched in Tasty Nucleotides

5'-nucleotides IMP and GMP (also known as Inosinate and Guanylate) are strong flavour enhancers (E626-E633) involved in "umami" taste. Since yeasts are rich in ribonucleic acid (RNA), a natural source of 5' nucleotides (GMP, AMP, CMP and UMP), yeast extracts enriched in 5' nucleotides IMP and GMP are currently used as taste enhancers.

5' nucleotide degradation yeast extract

5'-Nucleotidases and phosphatases are ubiquitous hydrolytic enzymes that catalyze the hydrolysis of nucleotides into a phosphate and nucleosides deprived of flavour-enhancing properties. The nucleosides are further hydrolyzed to purine and pyrimidine bases. Purine base hypoxanthine has a strongly bitter taste. Nucleotides/nucleosides/bases content in yeast extract vary as a function of growth state and the method of preparation.

Quality Control of Yeast Extract Nucleotides: NOVOCIB offers a range of analytical services from characterisation of free 5' nucleotides to full spectrum of nucleotides in yeast extracts by ion-pair high-performance liquid chromatography with diode array detection. This service allows to control the content of flavor nucleotides yeast extract samples.

Analytical system:

Agilent 1100 series liquid chromatograph fitted with binary pump, vacuum degasser, well-plate autosampler, thermcostatted column compartment and multiple wavelenght and diode array detector.
Run and data acquisision are controlled by Agilent Chem Station software. Zorbax Extend-C18 4.6x150mm, 3.5μm particle size and guard column (Agilent).
Calibrations are performed with standards prepared in mobile phase and with standards mixed with cell extracts, which are run immediately before and after every series of samples.
Peak assignment of different bases, ribonucleosides and ribonucleoside monophosphatesis is done by comparing both retention times and characteristics of UV absorption spectra (254/280 ratio) with those of standards. The area of individual peaks was measured using ChemStation software (Agilent).

inefficient yeast AMP demaination

 

References:
1. Vinas P, Campillo N, Melgarejo GF, Vasallo MI, Lopez-Garcia I, Hernandez-Cordoba M (2010) Ion-pair high-performance liquid chromatography with diode array detection coupled to dual electrospray atmospheric pressure chemical ionization time-offlight mass spectrometry for the determination of nucleotides in baby foods. J Chromatogr A 1217:5197–5203
2. Gill BD, Indyk HE (2007) Development and application of a liquid chromatographic method for analysis of nucleotides and nucleosides in milk and infant formulas. Int Dairy J 17:596–605
3. Yamaoka N, Kudo Y, Inazawa K, Inagawa S, Yasuda M, Mawatari K, Nakagomi K, Kaneko K (2010) Simultaneous determination of nucleosides and nucleotides in dietary foods and beverages using ion-pairing liquid chromatography-electrospray ionization-mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 878:2054–2060
4. Yang FQ, Li DQ, Feng K, Hu DJ, Li SP (2010) Determination of nucleotides, nucleosides and their transformation products in Cordycepsin Cordyceps by ion-pairing reversed-phase liquid chromatographymass spectrometry. J Chromatogr A 1217:5501–5510
5. Cichna M, Raab M, Daxecker H, Griesmacher A, Muller MM, Markl P (2003) Determination of fifteen nucleotides in cultured human mononuclear blood and umbilical vein endothelial cells by solvent generated ion-pair chromatography. J Chromatogr B Anal Technol Biomed Life Sci 787:381–391