Highly Sensitive In-Capillary Derivatization and Field Amplified Sample Stacking to Analyze Narcotic Drugs in Human Serum by Capillary Zone Electrophoresis

Separations

article

Monika Malak¹  Hager Ebrahim¹  Heba Sonbol¹  Ahmed Ali²  Yasmine Aboulella²  Ghada Hadad⁴  Samy Emara¹ 

[1]Pharmaceutical Chemistry Department, Faculty of Pharmacy, Misr International University

[2]Research Center, Misr International University

[3]Division of Systems Biomedicine and Pharmacology, Leiden Academic Centre for Drug Research ,(LACDR), Leiden University

[4]Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University

关键词: morphine;    morphine-6-glucuronide;    codeine;    field amplified sample stacking;    in-capillary derivatization;    capillary zone electrophoresis;    human serum;   

DOI  :  10.3390/separations10010058

学科分类：社会科学、人文和艺术（综合）

来源: mdpi

【 摘 要 】

An in-capillary derivatization (in-CAP-D) integrated with field amplified sample stacking (FASS) has been developed for the determination of morphine (MOR) and its metabolite, morphine-6-glucuronide (MOR-6-G) in human serum using capillary zone electrophoresis (CZE) and fluorescence detection (in-Cap-D-FASS-CZE). Acetonitrile was employed in removing proteins and extracting MOR and MOR-6-G into the clear supernatant containing codeine (COD) as an internal standard (IS). The derivatization was achieved in an in-capillary mode by introducing the acetonitrile-treated samples into a running electrolyte containing an oxidizing agent of potassium ferricyanide, whereas MOR, MOR-6-G and COD were oxidized into dimer derivatives with highly fluorescent intensity. The effectiveness and sensitivity of the in-Cap-D-FASS-CZE method were affected by many parameters, and the following conditions were found to be optimal: 70 m Mdisodium tetraboratedecahydrate (pH, 10.5), 0.30 mM ferrricyanide and a separation voltage of 10 kV. In order to perform the FASS, samples were electrokinetically injected for 20 s at 20 kV into the capillary that was pre-field with a 4 s water plug. Analysis was performed at ambient temperature (22 ± 1 °C). The method’s validation revealed good linearity with respect to peak area ratios of MOR and MOR-6-G with the IS and the corresponding concentrations over the ranges of 1–2000 and 1.2 to 2000 ng/mL, respectively. Following one oral dose of controlled-release MOR sulphate tablet, the validated in-Cap-D-FASS-CZE method successfully enabled the determination of MOR and MOR-6-G in clinical serum samples.

【 授权许可】

CC BY