Re: Colour of decalcifying fluid
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of Marvin Hanna) |
To: | histonet@histosearch.com |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
On Wed, 19 Jan 2000, A. F. Brandwood wrote:
> method of decalcification. In the past we used 5% Sulphosalicylic acid only
> to decalcify bone and the fluid turned purple with some of the bones.We now
> start off with 10% Nitric acid for a maximum of 48 hours and if the bone is
> not decalcified we transfer it to 5% Sulphosalicylic acid until
> decalcification is complete and we still get the purple colour with some
> bones.
According to the Merck Index, sulphosalicylic acid forms
a violet complex with ferric iron, and is used in a
colorimetric assay for Fe3+.
Possibly your 10% nitric acid treatment is releasing
"masked" iron from your bones, and the iron is then
chelated by the sulphosalicylic acid to form a
soluble coloured complex. If the specimens have spent
several weeks in non-neutralized formaldehyde prior to
decalcification, some of the haemoglobin will have
been converted to formalin pigment, from which iron
could be released by acid hydrolysis. Just a
suggestion; I don't think you mentioned the fixative
or the time.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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