Hemoglobin measurement at 540nm or 430nm?
gasmith at mail.med.upenn.edu
Sun Feb 9 12:33:28 EST 1997
In article <32F2151B.63E1 at biol.rug.nl>, Leendert Hamoen
<l.w.hamoen at biol.rug.nl> wrote:
> According to some literature the release of hemoglobin from erythrocytes
> after the addition of surfactants is measured at 540 nm. We measure a
> much higher absorption at 430 nm compared to 540 which is consistent
> with the fact that hemoglobin is red. Does anybody know why I should
> measure hemoglobin release at 540 nm in stead of 430 nm?
> Thanks in advance, Leendert Hamoen
In my experience, 430 nm is often TOO sensitive. Typical samples would
have to be diluted to get a reading that falls in the linear range. 540
nm is thus more convenient. I'd use 430 nm if I had very dilute samples,
but first I'd check that I was reading in the linear portion of the
absorbance vs concentration curve.
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