zeaxanthin et al.

David R Gerty dgerty at draper.com
Fri Jan 20 12:49:29 EST 1995


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



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