柱前衍生化-HPLC法测定人血浆中多胺浓度及其临床应用  

Determination of polyamines in human plasma by HPLC and its clinical application

作　　者：高梅梅[1] 李文龙[1] 耿春梅[1] 孔祥麟[1] 郭瑞臣[1] GAO Meimei LI Wenlong GENG Chunmei KONG Xianglin GUO Ruichen(Institute of Clinical Pharmacology, Qilu Hospital of Shanclong University, Jinan 250012, China)

机构地区：[1]山东大学齐鲁医院临床药理研究所,济南250012

出　　处：《中国临床药学杂志》2016年第6期344-348,共5页Chinese Journal of Clinical Pharmacy

摘　　要：目的建立同时测定血浆腐胺、精胺、亚精胺含量的柱前衍生化-HPLC法,并用于系统性红斑狼疮患者和正常人群血浆浓度测定,指导临床实践。方法 32例系统性红斑狼疮患者和30例健康志愿者血浆样品离心后,经丹磺酰氯衍生化和甲苯萃取,以C_(18)(250 mm×4.6 mm,5μm)柱为固定相,乙腈-0.2 mol·L^(-1)甲酸铵为流动相,经梯度洗脱,于272 mm波长处检测。结果腐胺、亚精胺质量浓度在0.2~40 mg·L^(-1)、精胺在0.2~30 mg·L^(-1)内呈现良好线性关系,腐胺、亚精胺最低定量限均为0.1 mg·L^(-1)、精胺为0.2 mg·L^(-1),腐胺、亚精胺最低检测限均为0.05 mg·L^(-1)、精胺为0.1 mg·L^(-1)。正常人群血浆腐胺浓度平均为1.570 mg·L^(-1)、精胺0.603 mg·L^(-1)、亚精胺1.710 mg·L^(-1)。结论建立了同时快速测定血浆中腐胺、精胺、亚精胺的柱前衍生化-HPLC法,方法灵敏、稳定、重现性好、选择性好、准确度高,可用于准确测定人血浆中多胺含量,并作为相关疾病诊断和治疗的重要参考指标。AIM To develop an HPLC method for determination of polyamines including putrescine, spermine and spermidine in human plasma, and apply in patients with systemic lupus erythematosus （SLE） and normal population, and guide the clinical practice. METHODS Plasma of 32 SLE patients was extracted with toluene after dansyl chloride derivative reaction, and plasma of 30 healthy volunteers was also extracted with toluene after dansyl chloride derivative reaction respectively. C18 column （250 mm × 4.6 mm, 5 μm） was used as stationary phase , acetonitrile- 0.2 mol·L^-1 atmnonium formate was used as mobile phase, gradient elution in ocpress was used, and the detective wavelength was 272 nm. RESULTS Linear ranges of the three kinds of polyamines were as following respectively ： putrescine and spennidine were in 0.2 - 40 mg·L^-1, and spermine was in 0.2 - 30 mg·L^-1. In their own linear range, they showed good linear relaontis （r 〉 0.99）. The limit of quantification （LOQ）concentrations were as following respectively： putrescine and spermidine were 0.1 mg·L^-1, and spermine was 0.2 mg·L^-1. The limit of detection（LOD）concentrations were as following respectively： putrescine and spennidine were 0.05 mg·L^-1, and spermine was 0.1 mg·L^-1. The average concentration of putrescine in healthy volunteers was 1.570 mg·L^-1, spenmdine was 0. 603mg·L^-1 and spennine was 1. 710 mg·L^-1respectively. CONCLUSION The experiment has established an HPLC method as described to detect the three kinds of polyamines in plasma rapidly at the same time. The method is sensitive, stable, with good reproducibility, selectivity and high accuracy, and can be used to detect the concentration of polyamine in human plasma accurately. At the same time, the level of polyamine can be made as an important reference index for related diseases in diagnosis and treatment.

关 键 词：多胺 高效液相色谱法 衍生化 系统性红斑狼疮 

分 类 号：R969[医药卫生—药理学]