Seperation and determination of anticoagulant cumarins which have different therapeutical effects by HPLC

The separation and determination of some anticoagulants drugs like cumarins derivatives were done directly by high performance liquid chromatography reverse phase.We got two separate mobile phases in different percentages, the first one is water 5%: methanol 95%, and the second is water 20%: acetonitril 80% with PH = 4 for both mobile phases. The steady phase was a column of octadecyle 5 micrometre [250mm X 46mm] Nucleosil, then we have got the best results of separation and retention time and determination of the concentrations using different wave lengths of U. V as a detector, that there was a special maximum wave length for each cumarin derivative, where coefficient variation was between 0.04%-3.9% and the limit of detection was between 0.3-1.9 microgram/ml for studied cumarins depending on the mobile phase and cumarin derivative. The assay method was used to qualify the purity of studied cumarin compounds especially used in human uses and quantify and separate it of other compounds in addition to toxicity cases. This method is accurate and sensitive based on quantitative studies