高效液相色谱法测定盐酸平阳霉素软膏的血药浓度

HPLC determination of plasma concentration of pingyangmycin ointment

目的:建立人血浆中盐酸平阳霉素的HPLC测定方法,从而研究患者使用盐酸平阳霉素软膏后的透皮吸收程度。方法:500μL人血浆用150μL10%的高氯酸沉淀后高速离心,取上清液进样20μL进行分析。流动相为甲醇-乙腈-0.002mol·L-1戊烷磺酸钠(15∶10∶75),用1mol·L-1硫酸调pH3.5;色谱柱:ODS-C18(150mm×4.6mm,5μm);检测波长:254nm;流速:1.0mL·min-1。10名志愿受试的寻常性银屑病患者使用盐酸平阳霉素软膏后,用HPLC法测定血浆中药物浓度。结果:本实验建立的血浆中盐酸平阳霉素HPLC测定方法,血浆中杂质不干扰样品的测定,药物在0.20～0.5mg·L-1的范围内线性关系好;最低检测为0.20mg·L-1;回收率高于80%;高、中浓度的日间和日内变异系数均小于10.0%;低浓度的日间和日内变异系数均小于15.0%,符合生物样品分析要求。10名患者用药后血浆中盐酸平阳霉素的药物浓度均低于最低检测浓度。结论:受试者对盐酸平阳霉素的吸收在0.20mg·L-1以下,发生药物不良反应的可能性较小,本研究中10名志愿受试者亦未出现不良反应。

OBJECTIVE A rapid and specific method for the determination of Pingyangmycin As （bleomycin As） in plasma is set up and used for the monitor of pingyangmycin in patients. METHODS A150μL aliquot of 10% perchloric acid was added to 500μL of plasma. The sample was vortexed and centrifuged, and 20μL of the supernatant was analyzed by HPLC at 254 nm using a reverse-phase column ODS-C18 （150 mm × 4. 6 mm, 5μm）. Elution was carried out using methanol-acetonitrile-0. 002 mol ·μL^- 1 sodium pentanesulfonate. Ten patients with psoriasis disease were enrolled in this study and the pingyamgmycin ointment was applied to the target zone on the body. RESULTS The calibration curve was found to be linear over the range of 0. 20-5. 0 mg· L^-1 with a precision of （10% （RSD%） and recovery higher than 80%. The inter- and intra-day variances were below 15.0%. The method was used to determine the plasma pingyangmycin concentration in patients with psoriasis disease after smearing with pingyangmycin ointment. CONCLUSION The plasma concentration of bleomycin A5 was below the LOQ,which implied that adverse effects were not likely to happen.