CHIRALPAK^® HSA HPLC Column

CHIRALPAK HSA, which uses human serum albumin as the chiral selector, is highly selective for acidic racemates, preferably weak and strong acids, zwitterionic and non-protolytic (neutral) compounds. Phosphate buffers (normally 0.01-0.1M, pH 5-7) with addition of organic modifiers are used as mobile phases. Enantioselectivity and retention can be regulated by changing the mobile phase composition. However, the primary use of CHIRALPAK HSA is for fast drug/protein binding studies (1). To calculate the % protein binding, measure the retention time of an unretained compound (t₀) and the compound of interest (t_r) on the CHIRALPAK HSA column. Then use the capacity factor equation:
k = (t_r - t₀)/t_r
to calculate the % protein binding (P):
P = 100k/(k+1)

Different types of mobile phases can be used. A mobile phase consisting of 6% 2-propanol in 20 mM potassium phosphate buffer, pH 7.0 gives data in good agreement with literature data. The mobile phase conditions should be chosen to suit the drugs to be tested, i.e., for high protein binding drugs a mobile phase with higher eluting strength might be needed in order to reduce retention times.

(1) Goodman, A.; Gilman, A.G. The Pharmacological Basis of Therapeutics, 9^th Edition, McGraw-Hill: New York, 1996; pp 1712-1792.

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CHIRALPAK is a registered trademark of Daicel Corp.