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The Study of Methodology and Applications of Deternination Cisatracurim

Author: TianGang
Tutor: DingMin
School: Chongqing Medical University
Course: Clinical Laboratory Science
Keywords: electrochemiluminescence capillary electrophoresis high performance liquid chromatography cisatracurium plasma
CLC: R927.2
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract


Objective:1. To develope a method for determination of cisatracurium by electrochemiluminescence （ECL）.2. A method for determination of cisatracurium by capillary electrophoresis with electrochemiluminescence（ECL） was developed.3. To established a method for determination of plasma cisatracurium by high performance liquid chromatography with fluorescence detection （HPLC-FD）. The developed method was applied to investigate the pharmacokinetics of cisatracurium in children.Methods:1. Cisatracurium was analysed by ECL detection. Detection system was tri-electrode working system. A background electrolyte of 0.8 mmol/L tris（2,2’-bipyridyl）ruthenium（II） (TBR, Ru（bpy）32+) and 40 mmol/L bibrate sodium sulution （pH 9.6）. The sample was quantitated by external standard curve method.2. Cisatracurium was analysed by CE-ECL detection. The running buffer solution was 100 mmol/L tris-HCl buffer solution （pH 8.5）. Detection system was tri-electrode working system. The background electrolyte was 65 mmol/L phosphate buffer （PBS） pH 8.5 and 5.0 mmol/L of Ru（bpy）32+ solution. Electrophoresis was carried out at 25℃. Cisatracurium was quantitated by internal standard curve using vecuronium bromide as internal standard.3. The chromatographic separation was performed on Shim-pack VP-ODS chromatography column （150 mm×4.6 mm, 5μm） at 25℃. The mobile phase was consisted with 30 mmol/L PBS （pH 3.0） containing 35 % of methyl alcohol and 5% of acetonitrile （v/v） at the flow rate of 0.8 mL/min from 0 to 3.5 min, 0.5 mL/ min from 3.5～6.0 min and 1 mL/min from 6.0～10 min. The analytes were monitored by a fluorophotometer at 236 nm （λex） and 324 nm （λem）. Cisatracurium in plasma was quantitated by internal standard curve using verapamil hydrochloride as internal standard.Results:1.The ECL assay of cisatracurium was linear in the range 1.25～60.0 mg/L, correlation coefficient for linear regression was 0.999 4, the detection limit was 0.050 mg/L. The average recoveries were 91.7%～98.2%. The average content of cisatracurium besylate for injection was （90.0±0.2）% （n=3）.2. CE- ECL assay was linear in the range 4.63～148.0 mg/L, correlation coefficient for linear regression was 0.998 7. The detection limit of cisatracurium was 0.46 mg/L. The average recoveries were 79.7%～96.9%. The average content of cisatracurium besylate for injection was （90.0±1.0）% （n=3）. 3. HPLC-FD assay was linear in the range 0.0 50～2.80 mg/L of cisatracurium in serum, correlation coefficient for linear regression was 0.999 7. The detection limit of cisatracurium was 12μg/L. The within-day coefficients of variation were lesss than 2% and the between-day coefficients of variation were less than 4%. The average recoveries were 93.3%～101.1%.4. The pharmacokinetics of cisatracurium in children after intravenous injection appeared to fit two compartments model. The primary pharmacokinetic parameters of cisatracurium in children were （ 32416±2649）μg·L-1·min for AUC∞, （ 3.67±0.60） min for T1/2a, （ 46.44±3.87 ） min for T1/2β, （ 2.08±0.32） mL·kg-1 for CL, （ 31.0±2.4 ）mL·kg-1·min-1 for Vd and （79.49±3.18） % for F, respectively.Conclusion:1. A novel ECL detection method for determination of cisatracurium has been develpoed. This method was simple and sensitive, suitable for the detection of cisatracurium besylate for injection .2. A new method was developed for determination of cisatracurium by CE-ECL detection. This method was sensitive, reliable and with wide linear range, suitable for the detection of cisatracurium besylate for injection .3. The developed methold for determination of cisatracurium in plasma by HPLC-FD was simple, sensitive, accurate and with broad linear range. The method was successfully applied to study the pharmacokinetics of cisatracurium in children after intravenous administration.

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CLC: > Medicine, health > Pharmacy > Pharmacopoeia Codex ( formulary ),drug identification > Drug identification > Determination of drug content
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