In article <Pine.3.89.9412201216.B10305-0100000 at uhunix.uhcc.Hawaii.Edu> yamamoto at UHUNIX.UHCC.HAWAII.EDU (Harry Yamamoto) writes:
>From: yamamoto at UHUNIX.UHCC.HAWAII.EDU (Harry Yamamoto)
>Subject: Re: zeaxanthin et al.
>Date: 20 Dec 1994 14:54:06 -0800
>The reference is:
>Gilmore AM and Yamamoto HY (1991) J. Chromatogr 543: 137-145.
>Thanks for the complement. I'm glad that people are finding the method
>useful.
>Harry Y. Yamamoto
>On Tue, 20 Dec 1994, Darren Goetze wrote:
>> actually xanthophyll cycle pigments can also be separated on plates as
>> well (see photosynthetica, v24,no3,p412 (1990)). however, in my
>> experience you are best off with a system that can achieve a real
>> separation of lutein and zeaxanthin (no mean feat; they are positional
>> isomers!). i have adapted the excellent protocol of gilmore and yamamoto
>> (j chromatography 1991, but i don't have the ref handy) and had superb
>> results, virtual baseline separation.
>> hope this helps,
>> best regards,
>> darren goetze
>> =========
>> dept botany
>> univ british columbia
>> vancouver bc
>> canada
>>>> On 19 Dec 1994, albino maggio wrote:
>>>> >
>> > I am looking for methods to measure levels of
>> > zeaxanthin, violaxanthin and b-carotene.
>> > Is HPLC the only way to separate them?
>> > What about quantification?
>> > Any reference or suggestion is welcome.
>> > Thanks in advance.
>> > Albino Maggio
>> >
I found that reverse phase hplc using a 20 minute gradient elution from 50%
MeOH/water to 100% MeOH worked very well for separating xanthophylls
quantitatively. Carotene was too slow to come off, a ternary solvent would
have helped. The support was 5 micron C-18. At the time I was looking at
chloropyllides and chlorophylls a,b and c, so was willing to compromise on the
carotenes.
A cellulose powder column and pet ether worked well for semi-preparative
work at around 30 psi.
This was for an MS thesis in 1979. I started with S. W. Jeffrey's two
dimensional thin layer chromatography technique on cellulose plates, but could
not get good quantitative results.
dgerty at draper.com
