Abstract
This paper describes a simple and sensitive high-performance liquid chromatographic method for measuring adenosine, inosine and hypoxanthine in cell suspensions. The method involves direct injection of the filtered sample on a microbore C18 reversed-phase column with UV detection at 259 nm. The mobile phase consisted of 125 mM potassium dihydrogenphosphate, 1.0 mM tetrabutylammonium hydrogen-sulfate, 1.5% acetonitrile and 20 mM triethylamine, pH 6.5. The minimum detectable amounts (signal-to-noise ratio of 3:1) were 2.0 pmol of adenosine, 2.5 pmol inosine and 3.5 pmol of hypoxanthine. The limits of quantitation were 2.9 ± 0.2 pmol for adenosine, 4.2 ± 0.3 pmol for inosine and 4,9 ± 0.4 pmol for hypoxanthine. This method was used to quantitate adenosine release by dispersed rat renal outer medullary cells (tubules) under conditions of normoxia and hypoxia.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 265-272 |
| Number of pages | 8 |
| Journal | Journal of Chromatography A |
| Volume | 536 |
| Issue number | C |
| DOIs | |
| State | Published - 1991 |
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ASJC Scopus subject areas
- Analytical Chemistry
- Clinical Biochemistry
- Molecular Medicine
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Quantitation of adenosine, inosine and hypoxanthine in biological samples by microbore-column isocratic high-performance liquid chromatography. / Gayden, R. H.; Watts, Bruns; Beach, R. E.; Benedict, C. R.
In: Journal of Chromatography A, Vol. 536, No. C, 1991, p. 265-272.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quantitation of adenosine, inosine and hypoxanthine in biological samples by microbore-column isocratic high-performance liquid chromatography
AU - Gayden, R. H.
AU - Watts, Bruns
AU - Beach, R. E.
AU - Benedict, C. R.
PY - 1991
Y1 - 1991
N2 - This paper describes a simple and sensitive high-performance liquid chromatographic method for measuring adenosine, inosine and hypoxanthine in cell suspensions. The method involves direct injection of the filtered sample on a microbore C18 reversed-phase column with UV detection at 259 nm. The mobile phase consisted of 125 mM potassium dihydrogenphosphate, 1.0 mM tetrabutylammonium hydrogen-sulfate, 1.5% acetonitrile and 20 mM triethylamine, pH 6.5. The minimum detectable amounts (signal-to-noise ratio of 3:1) were 2.0 pmol of adenosine, 2.5 pmol inosine and 3.5 pmol of hypoxanthine. The limits of quantitation were 2.9 ± 0.2 pmol for adenosine, 4.2 ± 0.3 pmol for inosine and 4,9 ± 0.4 pmol for hypoxanthine. This method was used to quantitate adenosine release by dispersed rat renal outer medullary cells (tubules) under conditions of normoxia and hypoxia.
AB - This paper describes a simple and sensitive high-performance liquid chromatographic method for measuring adenosine, inosine and hypoxanthine in cell suspensions. The method involves direct injection of the filtered sample on a microbore C18 reversed-phase column with UV detection at 259 nm. The mobile phase consisted of 125 mM potassium dihydrogenphosphate, 1.0 mM tetrabutylammonium hydrogen-sulfate, 1.5% acetonitrile and 20 mM triethylamine, pH 6.5. The minimum detectable amounts (signal-to-noise ratio of 3:1) were 2.0 pmol of adenosine, 2.5 pmol inosine and 3.5 pmol of hypoxanthine. The limits of quantitation were 2.9 ± 0.2 pmol for adenosine, 4.2 ± 0.3 pmol for inosine and 4,9 ± 0.4 pmol for hypoxanthine. This method was used to quantitate adenosine release by dispersed rat renal outer medullary cells (tubules) under conditions of normoxia and hypoxia.
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U2 - 10.1016/S0021-9673(01)89259-1
DO - 10.1016/S0021-9673(01)89259-1
M3 - Article
C2 - 2050767
AN - SCOPUS:0025961426
VL - 536
SP - 265
EP - 272
JO - Journal of Chromatography
JF - Journal of Chromatography
SN - 0021-9673
IS - C
ER -