Study on detection method of amphotericin B blood concentratio based on 2D-LC/UV
Objective Establish a rapid and accurate detection method for amphotericin B using two-dimensional liquid chromatography(2D-LC/UV)and apply the established method to monitor the blood concentration of amphotericin B in clinical patients.Methods This study was based on a new two-dimensional liquid chromatograph(2D-LC/UV).Plasma samples were processed by deproteinizing agents.On line solid phase extraction of amphotericin B in plasma was performed u-sing a one-dimensional chromatographic column SX1-1A(3.5mm ×25 mm,5 μm)to achieve preliminary enrichment and separation.Subsequently,it was transferred to a two-dimensional chromatographic column,Aston SCB column(4.6 mm × 125 mm,5 μm),for further separation and detection of the target analytes.The one-dimensional mobile phase was CAA-1D acidic mobile phase,with a flow rate of 0.7 mL/min;the two-dimensional mobile phase is BPI-3 alkaline mobile phase,with a flow rate of 1.0 mL/min.UV detection wavelength:380nm;Column temperature:40℃;after verifying the se-lectivity,linear range and lower limit of quantification,precision and recovery,matrix effect and stability of the method,the method was applied to the monitoring of amphotericin B blood concentration in clinical patients.Results The standard curve equation of amphotericin B in plasma was(y)=7204.6+914550x(R2=0.9999),There was a good linear relationship between concentration and peak area within the range of 0.07-12.31 μg/mL,and the lower limit of quantification was 0.07 μg/mL;the relative recoveries of low,medium,and high concentrations were between 97.4％and 105.3％,and the intra-day and inter-day precision were all below 6.1％,indicating good detection performance of this method.Serum samples were collect-ed from 20 patients using amphotericin B,and the patients'blood drug concentration ranged from 0.25 to 0.98 μg/mL.Conclusion The method is simple,fast,accurate and efficient,it realizes automatic analysis,and is suitable for high-throughput analysis and monitoring of clinical blood drug concentration;individual differences in patients using amphotericin B are large,and blood drug concentration monitoring is required during individualized treatment.
Two-dimensional liquid chromatographyAmphotericin BTherapeutic drug monitoring
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