Retention of Deoxyribonucleosides in HPLC with Reversed Phase. A Novel Relationship between the Capacity Factor and Organic Modifier Content

초록

The chromatographic retention behavior of five deoxyribonucleosides (dCyd, dUrd, dGuo, dThd and dAdo) was studied with respect to the mobile phase composition under isocratic conditions in reversed-phase HPLC. The Waters column packed by 10㎛ μ-Bondapak C18 was used. The volumn fraction(F) of the organic modifier was changed from 0 to 0.03, and to 0.12 for methanol and acetonitrile, respectively. Linear regression has been carried out according to the equations: log k'=log k'w-S*F (1), and k'=A+B*(1/F) (2) where k' is the solute capacity factor, and k'w, S,A, and B are experimental coefficients. The k'w values extrapolated from the experimental dependences of well known eqn.(1) do not coincide with the k'w values measured directly with a pure water as mobile phase. For example, the ratio of experimental and extrapolated k'w values for dAdo is more than 4. The novel relationship between the k' and F was proposed in. This equation has used an assumption that organic modifier and water adsorption on RP-HPLC material is described by Langmuir isotherm. It has been obtained that the correlation coefficients are always higher than 0.990 for (1) and (2). The slope ratios Bmethanol/Bacetonitrile and Sacetonitrile/Smethanol are changed for solutes in narrow ranges with averge values of 2.86 and 2.30, respectively, properties of solute and organic modifier simultaneously.