Separation and quantification of alkylphosphocholines by reversed phase high performance liquid chromatography

Alkylphosphocholines represent a new class of drugs with remarkable antineo plastic and antiprotozoal activity. For instance, hexadecylphosphocholine h as been approved for the topical treatment of skin metastasis. in addition, it was successfully studied in India for the treatment of leishmaniasis. D ifferent phase-I and phase-II-trials resulted in cure rates of more than 97 %. To optimize antitumor or antiprotozoal activity, we have prepared alkylp hosphocholines differing in chain length and unsaturation. For the qualitat ive and quantitative analysis of these longer chain analogues, we have used isocratic high performance liquid chromatography. The separation of the al kylphosphocholines with different chain lengths in this reversed phase HPLC system was achieved on a YMC-TMS column with a mobile phase consisting of methanol-water (85:15; v/v) at a flow rate of 1.0 ml/min. Furthermore the c is-/trans-isomers such as oleylphosphocholine and elaidylphosphocholine wer e clearly separated on a YMC-C8 column with a methanol-water mixture (80:20 ; v/v) as mobile phase. In the described reversed phase HPLC systems simple refractive index detection and UV detection allow the sensitive and quanti tative determination of alkylphosphocholines. These methods are very import ant for reproducible identification and quantitative determination of satur ated and mono-unsaturated alkylphosphocholines with alkyl residues containi ng up to 25 carbon atoms. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.