Total phospholipid assay?
j.broekman at verizon.net
Wed Mar 21 22:09:24 EST 2001
Wolfgang Schechinger wrote:
> first extract all lipids by Bligh and Dyer Method, then perform
> an essay for inorganic phosphate (molybdic acid procedure). If I
> remember correctly, there should be some papers from Schmitz-G and or
> Assmann and or Robenek from 1980 - 1990. Maybe you are able to
> locate them in PubMed. Actually, thre should be plenty of references,
> in that time and before that was common procedure.
> > From: ez <ezclimber at earthlink.net>
> > Subject: Total phospholipid assay?
> > Date: Wed, 21 Mar 2001 09:05:00 -0700
> > Organization: Posted via Supernews, http://www.supernews.com
> > To: methods at hgmp.mrc.ac.uk
> > Can anyone recommend an assay for total phosholipids in a complex
> > sample (natural biological material)?
> > We'd prefer easy-to-use (such as a dye blot) and very sensitive. But
> > anything will help at this point.
> > Thanks for helping!
> > Dave
> Dr. Wolfgang Schechinger
> Lab N233 (c/o Dr. Steve Roffler)
> Institute of Biomedical Sciences
> Academia Sinica
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> wolfsc at REMOVETHESEANTISPAMCAPITALSibms.sinica.edu.tw
This will NOT work. You have to digest/oxidize the PL to liberate the
Pi. Nevertheless, Bligh & Dyer is good. I have done many destructions
of dried phospholipids, using 70 % HClO4, 180°C, in a test tube with a
marble on top as condenser. You do want to do this in a well designed
fume hood, approved for HClO4. Failure to totally evaporate MeOH (or
other organics before heating the sample with HClO4 will give an
As starting point for a literature search (shameless self promotion):
Broekman, M. J. (1987) Ann.N.Y.Acad.Sci. 494, 95-97
Broekman, M. J., Handin, R. I., Derksen, A., and Cohen, P. (1976) Blood
Cohen, P., Broekman, M. J., Verkley, A., Lisman, J. W., and Derksen, A.
(1971) J.Clin.Invest. 50, 762-772
Broekman, M. J., Ward, J. W., and Marcus, A. J. (1980) J.Clin.Invest.
All this was LONG ago!
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