Wiczling Paweł, Kawczak Piotr, Nasal Antoni, Kaliszan Roman
Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gdañsk, Gen. J. Hallera 107, 80-416 Gdañsk, Poland.
Anal Chem. 2006 Jan 1;78(1):239-49. doi: 10.1021/ac0512103.
High-performance methods of testing of drug candidates for properties of pharmacokinetics and pharmacodynamics importance, in particular lipophilicity and acidity, are necessary to overcome innovation stagnation in the pharmaceutical industry. Reversed-phase high-performance liquid chromatography (RP HPLC) might be a unique tool for the determination of both pKa and the apparent (pH-dependent) partition coefficient, applicable in high-throughput analysis of multicomponent mixtures, e.g., samples originating from automated synthesis. In this work, the pH/organic modifier gradient RP HPLC is presented as a means of simultaneous determination of an analyte's acidity and lipophilicity. The approach consists of retention measurements in a series of methanol gradient runs differing in pH range and duration of the gradient. Two different models of the influence of pH on retention in organic modifier gradient RP HPLC are compared regarding the quality of the simultaneously determined lipophilicity and dissociation constants. Advantages of the proposed approach over currently employed procedures are that it can be applied to compound mixtures, it requires only minute amounts of substances, and pKa values can be determined in the range 3-10 units and lipophilicity in the range 0-7 units. Verification of the reliability of the parameters determined by the new method was demonstrated on a series of 93 acidic and basic drug analytes.
为克服制药行业创新停滞的问题，需要采用高效的方法来测试候选药物的药代动力学和药效学特性，特别是亲脂性和酸度。反相高效液相色谱法（RP HPLC）可能是一种独特的工具，可用于测定pKa和表观（pH依赖性）分配系数，适用于多组分混合物的高通量分析，例如来自自动合成的样品。在这项工作中，pH/有机改性剂梯度RP HPLC被作为一种同时测定分析物酸度和亲脂性的方法提出。该方法包括在一系列甲醇梯度运行中进行保留测量，这些运行的pH范围和梯度持续时间不同。比较了两种不同的pH对有机改性剂梯度RP HPLC中保留影响的模型在同时测定的亲脂性和解离常数质量方面的差异。所提出的方法相对于目前采用的程序的优点在于，它可应用于化合物混合物，仅需微量物质，并且pKa值可在3至10个单位范围内测定，亲脂性可在0至7个单位范围内测定。通过对一系列93种酸性和碱性药物分析物的测试，证明了新方法所测定参数的可靠性。
