Fast simultaneous determination of 14 nucleosides and nucleobases in cultured Cordyceps using ultra-performance liquid chromatography.

Determination of nucleosides and their metabolic compounds is important for physiological and pharmacological studies. Herein, a rapid ultra-performance liquid chromatography (UPLC) method was developed for the simultaneous determination of 14 nucleosides and nucleobases, namely adenine, adenosine, cytosine, cytidine, uracil, uridine, guanine, guanosine, hypoxanthin, inosine, thymine, thymidine, 2'-deoxyuridine and cordycepin. The separation was performed on Waters Acquity UPLC system with Acquity UPLC BEH C(18) column and gradient elution of 0.5mM acetic acid and acetonitrile in 5min. The correlation coefficients of 14 analytes were high (R(2)>0.9995) within the test ranges. The LOD and LOQ were lower to 11.9 and 47.0ng/ml with 1mul of injection volume, respectively. The overall R.S.D. for intra- and inter-day of 14 analytes were less than 1.8%. The developed method was applied for the analysis of nucleosides and nucleobases in cultured Cordyceps, which also could be used for the fast determination of the analytes in pharmaceutical products and biological fluids.

[1]  S. Hansen,et al.  Separation of nucleosides using capillary electrochromatography. , 1999, Journal of chromatography. A.

[2]  W. Völkel,et al.  Simultaneous determination of O6-methyl-2'-deoxyguanosine, 8-oxo-7,8-dihydro-2'-deoxyguanosine, and 1,N6-etheno-2'-deoxyadenosine in DNA using on-line sample preparation by HPLC column switching coupled to ESI-MS/MS. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[3]  G. Björk,et al.  Urinary excretion of pseudouridine and prognosis of patients with malignant lymphoma. , 1995, Acta oncologica.

[4]  G. Luo,et al.  Simultaneous determination of nine components in Qingkailing injection by HPLC/ELSD/DAD and its application to the quality control. , 2006, Journal of pharmaceutical and biomedical analysis.

[5]  P. Morin,et al.  Determination at ppb level of an anti-human immunodeficiency virus nucleoside drug by capillary electrophoresis-electrospray ionization tandem mass spectrometry. , 2000, Journal of chromatography. A.

[6]  T. Dong,et al.  Simultaneous determination of ergosterol, nucleosides and their bases from natural and cultured Cordyceps by pressurised liquid extraction and high-performance liquid chromatography. , 2004, Journal of chromatography. A.

[7]  F. Q. Yang,et al.  Quality control of Cordyceps sinensis, a valued traditional Chinese medicine. , 2006, Journal of pharmaceutical and biomedical analysis.

[8]  M. Mizugaki,et al.  Urinary excretion patterns of modified nucleosides, pseudouridine and 1-methyladenosine, in healthy individuals. , 1993, Clinica chimica acta; international journal of clinical chemistry.

[9]  Min Liu,et al.  Studies on the chromatographic behavior of nucleosides and bases on p-tert-butyl-calix[8]arene-bonded silica gel stationary phase by HPLC. , 2004, Talanta.

[10]  K. Kawamura.  Kinetics and activation parameter analyses of hydrolysis and interconversion of 2',5'- and 3',5'-linked dinucleoside monophosphate at extremely high temperatures. , 2003, Biochimica et biophysica acta.

[11]  Y. Wang,et al.  Simultaneous determination of six main nucleosides and bases in natural and cultured Cordyceps by capillary electrophoresis. , 2004, Journal of chromatography. A.

[12]  P. Solich,et al.  Advantages of application of UPLC in pharmaceutical analysis. , 2006, Talanta.

[13]  C. C. Marvel,et al.  Altered RNA turnover in carcinogenesis , 1994, Molecular and chemical neuropathology.

[14]  H. Cheung,et al.  Identification and quantification of base and nucleoside markers in extracts of Ganoderma lucidum, Ganoderma japonicum and Ganoderma capsules by micellar electrokinetic chromatography. , 2001, Journal of chromatography. A.

[15]  S. Ishiwata,et al.  Relationship of urinary pseudouridine and 1-methyladenosine to activity of leukemia and lymphoma. , 1992, Clinica chimica acta; international journal of clinical chemistry.

[16]  Fengqing Yang,et al.  Qualitative and quantitative determination of nucleosides, bases and their analogues in natural and cultured Cordyceps by pressurized liquid extraction and high performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC–ESI–MS/MS) , 2006 .

[17]  J. Gommers-Ampt,et al.  Method for the analysis of oxidized nucleosides by gas chromatography/mass spectrometry. , 1993, Analytical biochemistry.

[18]  C. Speicher,et al.  Elevated nucleoside excretion by patients with nasopharyngeal carcinoma: Preliminary diagnostic/prognostic evaluations , 1982, Cancer.

[19]  J. Harmenberg,et al.  Urinary modified nucleosides as tumor markers in cancer of the urinary organs or female genital tract , 1986, Urological research.

[20]  Michael E. Swartz,et al.  UPLC™: An Introduction and Review , 2005 .

[21]  T. Dong,et al.  Determination of nucleosides in natural Cordyceps sinensis and cultured Cordyceps mycelia by capillary electrophoresis , 2001, Electrophoresis.