Nojavan Saeed, Bidarmanesh Tina, Mohammadi Ali, Yaripour Saeid
Faculty of Chemistry, Shahid Beheshti University, Evin, Tehran, Iran.
Pharmaceutical Quality Assurance Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Electrophoresis. 2016 Mar;37(5-6):826-33. doi: 10.1002/elps.201500555.
In the present study, for the first time electromembrane extraction followed by high performance liquid chromatography coupled with ultraviolet detection was optimized and validated for quantification of four gonadotropin-releasing hormone agonist anticancer peptides (alarelin, leuprolide, buserelin and triptorelin) in biological and aqueous samples. The parameters influencing electromigration were investigated and optimized. The membrane consists 95% of 1-octanol and 5% di-(2-ethylhexyl)-phosphate immobilized in the pores of a hollow fiber. A 20 V electrical field was applied to make the analytes migrate from sample solution with pH 7.0, through the supported liquid membrane into an acidic acceptor solution with pH 1.0 which was located inside the lumen of hollow fiber. Extraction recoveries in the range of 49 and 71% within 15 min extraction time were obtained in different biological matrices which resulted in preconcentration factors in the range of 82-118 and satisfactory repeatability (7.1 < RSD% < 19.8). The method offers good linearity (2.0-1000 ng/mL) with estimation of regression coefficient higher than 0.998. The procedure allows very low detection and quantitation limits of 0.2 and 0.6 ng/mL, respectively. Finally, it was applied to determination and quantification of peptides in human plasma and wastewater samples and satisfactory results were yielded.
在本研究中，首次对基于电膜萃取-高效液相色谱联用紫外检测技术进行了优化和验证，用于定量分析生物和水样中的四种促性腺激素释放激素激动剂抗癌肽（阿拉瑞林、亮丙瑞林、布舍瑞林和曲普瑞林）。研究并优化了影响电迁移的参数。该膜由95%的1-辛醇和5%的二（2-乙基己基）磷酸酯固定在中空纤维的孔中组成。施加20 V的电场，使分析物从pH 7.0的样品溶液中迁移，通过支撑液膜进入pH 1.0的酸性接受溶液中，该接受溶液位于中空纤维的内腔中。在不同生物基质中，15分钟萃取时间内的萃取回收率在49%至71%之间，预富集因子在82至118之间，重复性良好（7.1＜相对标准偏差%＜19.8）。该方法具有良好的线性（2.0-1000 ng/mL），回归系数估计值高于0.998。该方法的检测限和定量限分别低至0.2和0.6 ng/mL。最后，将其应用于人体血浆和废水样品中肽的测定和定量分析，结果令人满意。
