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| Comparison of EMIT and 2D-HPLC in the quantification of Carbamazepine in serum of patients with epilepsy |
| LIU Hang WANG Min JIN Lu ZHANG Jinping |
| Department of Pharmacy, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Jiangsu Province, Nanjing 210008, China |
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Abstract Objective To compare the results of Carbamazepine in serum of patients with epilepsy determined by enzyme-multiplied immunoassay technique (EMIT) and two-dimensional liquid phase chromatography(2D-HPLC). Methods The serum concentration of Carbamazepine in 67 epilepsy patients was determined by EMIT and 2D-HPLC, respectively. In 2D-HPLC method, two reversed phase liquid chromatography columns [ASTON-FAQ C8 (4.0 mm×100.0 mm, 3 μm, ANAX) and Diamonsil-C18 (4.6 mm×200.0 mm, 5 μm, Dikma) column] were used as the first and second separation dimensions, respectively. ASTON C18 (4.6 mm×10.0 mm, 3 μm, ANAX) column was used as a connector to transfer the eluent from the first dimensional column to the second column for further separation. The mobile phases of the two dimensions consisted of acetonitrile-H2O (55∶45, v/v) and acetonitrile-20 mmol/L ammonium acetate (20∶80, v/v, pH=7.0), respectively. The first dimensional column flow rate was 1.0 mL/min, the second dimensional column flow rate was 1.2 mL/min. The detection wavelength was 285 nm, and the column temperature was 40℃. Results There were no significant differences in the serum concentrations of Carbamazepine detected by EMIT and 2D-HPLC. A good correlation was obtained between the two methods. The regression equation was y(2D-HPLC)= 0.9876 × x(EMIT)-0.0313. Bland-Altman plots analysis showed the two methods were consistent. Conclusion Both EMIT and 2D-HPLC methods have good consistency and correlation in the quantification of Carbamazepine. The laboratory conditions and staffing should be taken into consideration when a method is chosen for monitoring the concentration of Carbamazepine.
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