A chiral HPLC method for the determination of low amounts of D-carnitine in L-carnitine after derivatization with (+)-FLEC.
An indirect enantioseparation method for robust and precise determination of D-Carnitine (D-C) in L-Carnitine (L-C) in the range of 0.1-1.0% is presented. The method is based on derivatization of Carnitine with (+)-[1-(9-fluorenyl)-ethyl]-chloroformate ((+)-FLEC). The two diastereomers are subsequently separated of on an octadecyl column using detection of the eluent by fluorescence (260 excitation, 310 nm emission monitoring). This procedure can be calibrated conveniently by diluting the derivatization solution of the sample. Hence, D-C was determined indirectly through quantification of L-C thereby strongly increasing the robustness and reducing the costs. During the development of the method a study was undertaken to prove that the method is suitable to determine enantiomeric purity of L-C indeed. Moreover, the method was validated according to the ICH guidance, which required the additional performance of a collaborative study. The proposed assay can be carried out using an autoinjector because the derivatives are very stable. Hence, we believe that this method will become popular for reliable determination of enantiomeric purity of L-C.
Freimüller
Sascha
S
Institute of Pharmaceutical Sciences, Federal Institute of Technology (ETH), 117 M48 Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
Altorfer
Hans
H
D002331:Carnitine / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000706:statistics & numerical data; D005449:Fluorenes / Q000032:analysis* / Q000737:chemistry; D013237:Stereoisomerism
D005449:Fluorenes; C052812:1-(9-fluorenyl)ethyl chloroformate; D002331:Carnitine