A simple and rapid high performance liquid chromatography (HPLC) method using ultraviolet (UV) detection was developed to determine hydroxyurea (HU) concentration in plasma sample after derivatization with xanthydrol.

Two hundred microliters samples were spiked with methylurea (MeU) as internal standard and proteins were precipitated by adding methanol. Derivatization of HU and MeU was immediately performed by adding 0.02 M xanthydrol and 1.5 M HCl in order to obtain xanthyl-derivatives of HU and MeU that can be further separated using HPLC and quantified using UV detection at 240 nm.

Separation was achieved using a C18 column with a mobile phase composed of 20 mM ammonium acetate and acetonitrile in gradient elution mode at a flow rate of 1 mL/min. The total analysis time did not exceed 18 min. The method was found linear from 5 to 400 μM and all validation parameters fulfilled the international requirements. Between- and within-run accuracy error ranged from −4.7% to 3.2% and precision was lower than 12.8%.

This simple method requires small volume samples and can be easily implemented in most clinical laboratories to develop pharmacokinetics studies of HU and to promote its therapeutic monitoring.

建立了一种使用紫外线（UV）检测的简单，快速的高效液相色谱（HPLC）方法，以测定在用Xanthydrol衍生化后血浆样品中的羟基脲（HU）浓度。

向200微升样品中加入甲基脲（MeU）作为内标物，并通过添加甲醇使蛋白质沉淀。立即通过添加0.02 M的黄嘌呤醇和1.5 M的HCl进行HU和MeU的衍生化，以获得HU和MeU的黄嘌呤衍生物，可以使用HPLC进一步分离它们，并使用UV检测在240 nm处进行定量。

使用带有流动相的C18色谱柱进行分离，该流动相由20 mM乙酸铵和乙腈组成，梯度洗脱模式，流速为1 mL / min。总分析时间不超过18分钟。发现该方法在5至400μM之间呈线性，并且所有验证参数均满足国际要求。批间和批间准确度误差在-4.7％到3.2％之间，并且精度低于12.8％。

这种简单的方法需要少量样品，并且可以在大多数临床实验室中轻松实施，以开展HU的药代动力学研究并促进其治疗监测。