Studies were carried out to develop an HPLC method for the determination of lanthanides, in presence of Th and U, without involving any pre-separation procedure for the matrix elements (Th, U). The effect of concentration of the ion interaction reagent on the retention pattern of lanthanides, thorium and uranium was studied using a reversed-phase C18 column. The chromatographic parameters were optimized for maximizing the capacity of the column to hold thorium. Dual gradient elution was used for the separation of lanthanides from a mixture of thorium and uranium present in large proportion. The method facilitates the use of a minor fission product as an internal standard for the accurate and precise quantification of lanthanides by HPLC. The developed methodology was validated by using synthetically prepared samples with different proportions of lanthanides. Studies were also carried out for the separation and determination of U and Th on a reversed-phase column. A gradient of methanol and alpha-hydroxy isobutyric acid was employed for the separation and determination of uranium in presence of large amounts of thorium.