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Dear All
We are working with drug distribution studies of an NCE's in Rat
skin, we would be thankful if any body can suggest a method for
homogenization of skin,
i.e regarding the compositon of the homogenizing buffer, pre-
homogenization procedure etc and we are trying to extract back drugs
delivered into skin from the skin itself.
Does anyone can help with a good homogenization method for a whole skin
Thanks, Regards
Shalini Malho
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Hello
As we are working on homogenization of various tissues and organs we
found the comfortable way of homogeniztion is by chopping the tissue
and later homoginate with the homogenizer (Bio fuge)or by ultra sonic
homogenizer.
I think you can try the same method by chopping the skin and
homogenate.
And you select the solvent or buffer in which the analyte can be
extracted back while homogenization use that solvent as an
homogenization solvent.
Regards
Nagesh
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Dear Shalini,
I have recently evaluated the fastprep system (MP biomedical) for the
homogenisation full thickness human and pig skin (approx. 1 cm2 surface
area). The results were very encouraging and I am in process of buying
one unit for our skin penetration studies (the solvent we used was a
simple water: methanol solution).
There are various lysing matrices available (see their website) and a
very abrasive one is required for skin.
Regards
Raniero
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
Unilever U.K. Central Resources Limited
Registered in England & Wales; Company No 29140
Registered Office: Unilever House, Blackfriars, London, EC4P 4BQ
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Does anyone else have experience using this system tissue DMPK studies?
The company's literature seems entirely directed towards DNA, RNA, and
protein isolation.
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Dear Mary
In the following free paper a good skin sample preparation is described
Satoshi Sobue, Kaneo Sekiguchi, and Toshitaka Nabeshima
Intracutaneous Distributions of Fluconazole, Itraconazole, and
Griseofulvin in Guinea Pigs and Binding to Human Stratum Corneum
Antimicrob. Agents Chemother. 2004 48: 216-223.
Regards
Valmen
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There is a rather low tech, but very effective mechanism of doing
this that involves liquid nitrogen pulverization of full thickness
skin, up to 2 cm squares in a precision fit piston using a hammer.
The resultant powder can be resuspended in a phosphate buffer and
subjected to an ST-10 Polytron probe (or turrax large tooth probe)
for about 30 seconds. This can then be extracted for your compound
using liquid liquid extractions. Alternately, you can use a pre-
chilled coffee grinder (available from any department store and kept
in a -20 freezer), sliver the skin, freeze it in liquid nitrogen,
grind it in a cold room and you have a supepndable or extractable
powder in less than a minute. Remember, no liquid Nitrogen in the
grinder, they do explode.
Sanjeev Thohan, PhD
Director, DMPK
Exelixis, Inc
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