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Determination of Morphine Concentration in Human Plasma by HPLC

Objective: To establish a method to determine rapidly morphine in human palsma by HPLC. Methods:The separation was performed on Waters Spherisorb C18 column with 0.05 mol/L potassium dihydrogen phosphate-0.002 5 mol/L sodium heptanesulfonate-acetonitrile(5:5:2) as mobile phase using perchloric acid as protein precipitant.The column temperature was 30 ℃, the flow rate was 0.5 m L/min, the detection wavelength was 220 nm, and the sample volume was 20 μL. Results: The calibration curve of morphine was linear within the range of 1.0~100 μg/m L with good linearity(r=0.999 3). The lowest detection limit was 0.3 μg/m L. The recovery was 98.70%, 113.71% and 96.51%with intra-day RSD of 3.39%, 4.96% and 2.27% and inter-day RSD of 7.64%, 5.74% and 3.18% for low, medium and high concentration, respectively. Conclusion: This method is simple, rapid and stable for the determination of morphine concentration in human plasma.