4. Discussion
Organic acids frequently represent unique chromatographic separation challenges, with strategies for increased
resolution limited to optimising column length, temperature, organic modi®er content, ionic strength or pH
adjustment of eluent in order to suppress both analyte dissociation and silanol ionisation. Retention under
reversed-phase conditions is generally the result of several interaction mechanisms between polar solute, eluent, stationary phase and underlying silica, with the consequence that column selection and optimisation are rarely facile Claessens, van
Recent HPLC methods for the determination of pantothenic acid in infant formulas have advocated reversed-
phase separation at both low pHand low organicmodirer
concentration It is recognised that both of these requirements, while essential for successful retention of
a polar and ionisable analyte, represent potential limitations to elective reversed-phase performance, espe-
cially with the older type A silica column chemistries.Thus, both conventional end-capping protection of
residual silanols and bonded-phase integrity are vulnerable to low pH, with variable selectivity commonly
reported under such conditions . Further, the predominantly aqueous eluents (>95%) required to e€ect retention of such polar analytes, can initiate collapse of the hydrophobic bonded phase, with potential variability, or loss of retention.
This problem has previously been described during pharmaceutical analysis (Woollard, 1986).
The six columns selected during column evaluation represent di€erent solutions to the above considerations.
The Resolve C18 cartridge column is representative of asrst generation, non-endcapped type A silica chemistry,
Table 1
Pantothenic acid content in milk and infant formula powders (mg/100
g)a