
%0 Journal Article
%A Naidu, Bodapati P. 
%T Separation of sugars, polyols, proline analogues, and betaines in stressed plant extracts by high performance liquid chromatography and quantification by ultra violet detection
%J Functional Plant Biology
%D 1998/12/21
%V 25
%N 7
%P 793-800
%K Osmoprotectants, solutes, cotton, peanut, Melaleuca.
%R https://doi.org/10.1071/PP97165
%U https://www.publish.csiro.au/paper/PP97165 
%X Osmoprotectants such as (i) sugars and sugar alcohols (polyols), (ii) proline and its analogues, and (iii) a number of quaternary ammonium compounds (betaines) play a significant role in plant adaptation to environmental stresses. Lack of a simple and rapid technique for the extraction and simultaneous determination of these osmoprotectant solutes from a large number of samples originating from plant breeding populations and agronomic trials, led to the development of this method. Osmoprotectants were extracted using methanol-chloroform-water. Extracts were partially purified, where required, by treating with ion exchange resins. Solutes were separated using a high performance liquid chromatograph fitted with a Sugarpak-1 column and 5 mg L-1 Ca-EDTA solution as the mobile phase. The eluted solutes were quantified by ultra violet detection at 195 nm. This technique measures sugars/polyols, proline analogues, and betaines in a variety of plant species such as peanut, Melaleuca uncinata, and cotton, respectively. Pinitol levels in peanut estimated using this method correlated significantly with the determinations obtained by HPLC-refractive index determination (R2 = 0.983, P&#60;0.001). Glycinebetaine determinations in cotton using this method were significantly correlated with determinations achieved by 1H NMR spectroscopy (R2 = 0.989, P&#60;0.001). Cotton leaf extracts spiked with a range of authentic glycinebetaine levels were also precisely measured using this technique (R2 =0.999, P&#60;0.001). The described method is simple, rapid, sensitive, cost effective and simultaneously measures more than one class of osmoprotectants from a single chromatographic run. 
