Reversed Phase HPLC Characterisation

Linear gradient from 10% v/v of B to 65% v/v of B in 45 minutes was used at a flow rate of 2 ml/min at ambient temperature and UV detection at 208 and 214 nm, respectively. Polymer samples were dissolved in eluent A at a concentration of 3mg/ml. Depending on the UV absorbance of the polymer 20 to lOOpl aliquots were injected. 

Both the basic and amphoteric polymers form highly charged TFA salt under the acidic elution conditions. Wide elution profile was observed only with slight differences in retention times. It is interesting to note that the polyanionic polymer (Ac-EAK) produced a relatively sharp peak. It might be due to die lack of salt formation with TFA. 

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Various new methods such as capillary electrophoresis and reversed-phase HPLC have been used to characterise glutenin subunits. These... 

Colored compounds were isolated and characterised from the reaction products of xylose (1M) and glycine (1M), refluxed for 2 h, initial pH 6. The solubles in light petroleum ether (b.p. 60-80°) were fractionated into at least 13 peaks by semipreparative reverse-phase HPLC, using a water-methanol gradient. 

Molnar and Horvath [140] proposed reversed-phase HPLC for characterisation of lipophilicity, by the use of log A- instead of log P. Later on, Biagi et al. 23] found a good correlation between log P. log A and Py/ values, determined by TLC. 

The ionised sulphate group makes carotenoid sulphates strongly polar. This results in significant solubility in water with values up to 0.4 mg/ml. The high polarity and solubility properties determine the choice of isolation procedure. Special methods are employed and detailed isolation procedures available [68]. The isolation methods include column chromatography on Sephadex or silica, TLC and reversed phase HPLC. Sulphur analysis is also desirable. Further characterisation is carried out by spectroscopic methods. 

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