Objective To establish a rapid assay for the determination of
methotrexate (MTX) in serum and separate the
enantiomers in
drugs by different manufactorys.Methods The assay base on
methotrexate has ultraviolet absorbability at 306nm,separation of the drug by high performance capillary electrophoresis(HPCE).The naked capillary is 60cm in length and efficient length is 50.5cm.The voltage is 25kv ,and the buffer is 75mmol/L phosphate additive,the methotrexate
enantiomers were completely separated by adding cyclodextrin in buffer.Results Complete separation of MTX was achieved within 10 minutes.The assay was linear from 1.1μmol/L to 1100.0μmol/L.The minimal detection limit was 0.55μmol/L.The recovery of MTX was 88.2%～98.2%.Wthin-run precision was 4.2%,and between-run precision was 5.4%.Additional,by added modifer and chiral reagent to separate the enantiomers.Conclusions HPCE is an effective method in clinical and scientific research for its advantages such as rapidity,simplicity and acccuaracy in determination of methotrexate in serum and separation of the enantiomers in drugs by different manufactorys.