Dr Engelbert Buxbaum engelbert_buxbaum at
Fri Jul 2 02:40:54 EST 2004

Martin wrote:

> I will be much gratefull if somebody tell how can I measure ATP.
> I have use luciferin-luciferase assay but would like to compare this
> method with others.

You could use a coupled spectrophotometric assay, if sample volume and
ATP-concentration are sufficiently high.

ATP, like all nucleotides, has a strong absorbtion at 258 nm, which can
be measured. If interfering substances are present, this can be done
following ion exchange or ion pairing chromatography (see for example
Ushimaru & Fukushima: Anal. Biochem 313 (2003) 173-5), using the area of
the chromatographic peak as measured variable.

Theoretically, it should also be possible to measure the concentration
of ATP, like any other compound, by MS or (if there may be compounds of
similar MW present) MS/MS. This would be a very sensitive method, if you
can get hold of the instrument.

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