Total phospholipid assay?

[Han Broekman]

Wolfgang Schechinger wrote:
> 
> Dave,
> 
> 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.
> 
> Wolfgang
> 
> > 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
> Yen-Chio Yuan Rd. Sec.2
> Taipei 115
> Taiwan R.O.C.
> Tel +886-2-2798-9152
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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
explosion.

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
47, 963-971
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.
66, 275-283

All this was LONG ago!
-- 
Best regards
Han Broekman
(Please answer to the newsgroup only, I will not answer email)