| Objective:Establish a high performance liquid chromatographic?HPLC?method for the determination of vancomycin in serum and synovial fluid,study vancomycin concentration in serum and synovial fluid after locally application in arthroplasty,and provide a reference for locally application of vancomycin in arthroplasty.Methods:A HPLC method was established for the determination of vancomycin in serum and synovial fluid by optimizing the determination conditions for HPLC and the treatment for blood and synovial fluid samples.Methodology was examined for linear range,lower limit of quantitation,specificity,precision,recovery and stability.From July 2017 to July 2018,patients locally applied vancomycin in arthroplasty was studied.Serum and synovial fluid samples were collected from patients and the concentration of vancomycin was determined by HPLC.Patients undergoing primary arthroplasty were divided into three groups according to the dosage of vancomycin.Group A:0.5 bottles of vancomycin powder?250mg?were evenly applied to the wound and bone surface;Group B:1 bottle of vancomycin powder?500mg?was evenly applied to the wound and bone surface;Group C:2 bottles of vancomycin powder?1000mg?were evenly applied to the wound and bone surface.Statistical analysis was performed by SPSS 22,and graphic drawing was performed by GraphPad Prism 5.Results:A linear regression equation of vancomycin in serum was determined by HPLC:y=3806.2x+48724,R2=0.998(from 5.00 to 80.00mg·L-1).The vancomycin lower limit of quantitation in serum was 5.13mg·L-1;A linear regression equation of vancomycin in synovial fluid was determined by HPLC:y=3806.1x+12297,R2=0.996(from 2.00 to 3000.00mg·L-1).The vancomycin lower limit of quantitation in synovial fluid was 2.03mg·L-1.Good specificity;The recovery ranged from 89.26%to106.50%,RSD<4%;The RSD of intra-day and inter-day were less than 8%;Serum and synovial fluid samples were placed under 20 degree centigrade for 0 hour,12hours,24 hours,48 hours,72 hours and repeated freezing and thawing 3 times under-80 degree centigrade,the RSD was less than 9%.The methodological study meets the requirements.In the study,286 samples were collected from 45 patients undergoing primary arthroplasty.The vancomycin concentration in serum was less than 5.00mg·L-1 at 3hours after locally applied vancomycin in primary arthroplasty?7 patients?.The method of collecting synovial fluid samples was studied.It have been found that the synovial fluid samples collected by b collecting method at 1 hour,3 hours,6 hours,12hours,18 hours and 24 hours was better.Group A:vancomycin concentration in synovial fluid at 1 hour,3 hours,6 hours,12 hours,18 hours and 24 hours after arthroplasty was 60.05±37.98mg·L-1,67.16±59.04mg·L-1,45.97±41.11mg·L-1,39.10±30.58mg·L-1,30.86±22.08mg·L-1,18.92±14.92mg·L-1?11 patients?;Group B:vancomycin concentration in synovial fluid at 1 hour,3 hours,6 hours,12 hours,18hours and 24 hours after arthroplasty was 521.39±179.51mg·L-1,247.96±118.81mg·L-1,164.25±64.44mg·L-1,174.74±82.61mg·L-1,127.57±96.44mg·L-1,83.00±47.99mg·L-1?16 patients?;Group C:vancomycin concentration in synovial fluid at 1 hour,3 hours,6 hours,12 hours,18 hours and 24 hours after arthroplasty was 1488.09±370.90mg·L-1,1054.39±715.25mg·L-1,917.03±698.20mg·L-1,581.49±361.67mg·L-1,232.12±164.23mg·L-1,114.47±86.90mg·L-1?9 patients?.The mixed effect model was used to analyze factors affecting vancomycin concentration in synovial fluid.Only time and the dosage of vancomycin correlated with the logarithm of the vancomycin concentration in synovial fluid.The regression coefficient of vancomycin dosage was??95%CI?=0.71?0.37,1.05?,P=0.001;the regression coefficient of time was??95%CI?=-0.024?-0.034,-0.015?,P<0.001.During the entire observation period,1 patient had periprosthetic joint infection after primary arthroplasty,and no adverse drug reaction occurred in all patients.Conclusion:The high performance liquid chromatography method established in this study is accurate and reliable in the determination of vancomycin concentration in serum and synovial fluid.Vancomycin has important clinical significance in the prevention of periprosthetic joint infection.Locally application of vancomycin powder in arthroplasty can produce a high vancomycin concentration environment in the joint cavity and effectively prevent periprosthetic joint infection.It yields low systemic vancomycin concentration,which has low risk of adverse reactions and high-level safety.The vancomycin concentration in synovial fluid only relates to time and the dosage of vancomycin.The half-life of vancomycin in the joint cavity is 13.3 hours.This study provides a reference for locally application of vancomycin in arthroplasty. |
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