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Mass spectrometry based analysis of nucleotides, nucleosides, and nucleobases—application to feed supplements

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Abstract

In this work, accurate MS-based methods for quantitative profiling of nucleotides, nucleosides, and nucleobases in yeast extracts used as additives in animal feedstuff are presented. Reversed-phase chromatography utilizing a stationary phase compatible with 100 % aqueous mobile phases resulted in superior analytical figures of merit than HILIC or ion-pair reversed-phase separation. The novel separation method was combined with both molecular and elemental mass spectrometry. By use of RP–LC–MS–MS, excellent limits of detection <1 μmol L−1 could be obtained for all the compounds investigated. The elemental speciation analysis approach enabled determination of nucleotides by phosphorus detection. Sensitivity of LC–ICP–MS was 1–2 orders of magnitude lower than that of LC–MS–MS. Quantitative analysis of yeast products using complementary MS detection furnished values in good agreement.

Reversed phase chromatographic separation of nucleotides, nucleosides and nucleobases combined with mass spectrometric detection

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Abbreviations

ACN:

Acetonitrile

ADP:

Adenosine diphosphate

AMP:

Adenosine monophosphate

As:

Adenosine

ATP:

Adenosine triphosphate

Cd:

Cytidine

CE:

Capillary electrophoresis

CMP:

Cytidine monophosphate

DNA:

Deoxyribonucleic acid

DRC-QMS:

Dynamic reaction cell–quadrupole mass spectrometry

ESI:

Electrospray ionization

FV:

Fragmentor voltage

GMP:

Guanosine monophosphate

Gs:

Guanosine

HILIC:

Hydrophilic-interaction liquid chromatography

HPLC:

High-performance liquid chromatography

IC:

Ion chromatography

ICP:

Inductively coupled plasma

IMP:

Inosine monophosphate

IP:

Ion pair

IS:

Internal standard

Is:

Inosine

k′:

Capacity factor

LC:

Liquid chromatography

LOD:

Limit of detection

MeOH:

Methanol

MRM:

Multiple reaction monitoring

MS-MS:

Tandem mass spectrometry

N:

Number of theoretical plates

NDP:

Nucleoside diphosphate

NMP:

Nucleoside monophosphate

NTP:

Nucleoside triphosphate

RP:

Reversed phase

tR:

Retention time

Ud:

Uridine

UMP:

Uridine monophosphate

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Acknowledgements

This work was supported by the Biomin B.R.A.I.N program and by the Austrian Research Promotion Agency FFG (FHplus projekt METORGANIC).

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Authors and Affiliations

  1. Department of Chemistry, Division of Analytical Chemistry, University of Natural Resources and Life Sciences-BOKU, 1180, Vienna, Austria

    Stefan Neubauer, Ariana Rugova, Dinh Binh Chu, Hedda Drexler, Stephan Hann & Gunda Koellensperger

  2. FH Campus Wien, Bioengineering, 1100, Vienna, Austria

    Stefan Neubauer & Michael Sauer

  3. Department of Chemistry, Faculty of Mathematical and Natural Sciences, University of Prishtina, 10 000, Pristina, Kosovo

    Ariana Rugova

  4. Biomin Research Center, 3430, Tulln, Austria

    Anja Ganner

  5. Department of Biotechnology, University of Natural Resources and Life Sciences-BOKU, 1180, Vienna, Austria

    Michael Sauer & Diethard Mattanovich

  6. Austrian Center of Industrial Biotechnology (ACIB), 1180, Vienna, Austria

    Michael Sauer, Diethard Mattanovich, Stephan Hann & Gunda Koellensperger

Authors
  1. Stefan Neubauer
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  2. Ariana Rugova
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  3. Dinh Binh Chu
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  4. Hedda Drexler
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  5. Anja Ganner
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  6. Michael Sauer
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  7. Diethard Mattanovich
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  8. Stephan Hann
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  9. Gunda Koellensperger
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Corresponding author

Correspondence to Gunda Koellensperger.

Additional information

Stefan Neubauer and Ariana Rugova contributed equally

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Neubauer, S., Rugova, A., Chu, D.B. et al. Mass spectrometry based analysis of nucleotides, nucleosides, and nucleobases—application to feed supplements. Anal Bioanal Chem 404, 799–808 (2012). https://doi.org/10.1007/s00216-012-6170-9

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  • DOI: https://doi.org/10.1007/s00216-012-6170-9

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