Simple Quantitation of Flucytosine in Low Volume Serum Samples and Evaluation of Common Endogenous Interferences by HPLC-UV  

作　　者：Abdul Rafiq Khan Ali Alothaim Waleed Tamimi Majid Alshaibani Souad Marwan Al Enazi Syed Muhammad Saad Khalid Mohammed Khan Abdul Rafiq Khan;Ali Alothaim;Waleed Tamimi;Majid Alshaibani;Souad Marwan Al Enazi;Syed Muhammad Saad;Khalid Mohammed Khan(Department of Pathology and Laboratory Medicine, King Abdulaziz Medical City, Riyadh, KSA;Department of Chemistry, University of Karachi, Karachi, Pakistan;H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan)

机构地区：[1]Department of Pathology and Laboratory Medicine, King Abdulaziz Medical City, Riyadh, KSA [2]Department of Chemistry, University of Karachi, Karachi, Pakistan [3]H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan

出　　处：《American Journal of Analytical Chemistry》2022年第9期289-299,共11页美国分析化学（英文）

摘　　要：5-Fluorocytosine (5-FC) is used for the treatment of several infections. It is extremely important to monitor blood level concentration for maximum activity to avoid its side effects. A simple, faster, and more accurate analytical method is developed and validated using high-performance liquid chromatography with UV detection in a very low-volume serum sample. Exactly 50 μL of serum was precipitated with 5% trichloroacetic acid. After mixing and centrifugation, 20 μL of supernatant was injected into the HPLC column. Detection was performed at 280 nm. The method is very specific and free from interfering substances due to different drugs and their different circulating metabolites. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.50 μg/L and 1.0 μg/L, respectively. The method was linear in the range of 5 - 150 μg/L in the serum sample. In method comparison, the correlation coefficient r² was 0.999 and the percentage recovery was 90% - 105% on four levels of the quality control samples. Within run and between run precision was found to be less than 2.2% at four different concentrations (5, 25, 50, and 100 μg/L). A simple, faster, and more accurate HPLC-UV method is developed which is very useful for monitoring 5-FC concentration in low volume serum samples without evaporation step and ion exchange chromatography within minutes.5-Fluorocytosine (5-FC) is used for the treatment of several infections. It is extremely important to monitor blood level concentration for maximum activity to avoid its side effects. A simple, faster, and more accurate analytical method is developed and validated using high-performance liquid chromatography with UV detection in a very low-volume serum sample. Exactly 50 μL of serum was precipitated with 5% trichloroacetic acid. After mixing and centrifugation, 20 μL of supernatant was injected into the HPLC column. Detection was performed at 280 nm. The method is very specific and free from interfering substances due to different drugs and their different circulating metabolites. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.50 μg/L and 1.0 μg/L, respectively. The method was linear in the range of 5 - 150 μg/L in the serum sample. In method comparison, the correlation coefficient r² was 0.999 and the percentage recovery was 90% - 105% on four levels of the quality control samples. Within run and between run precision was found to be less than 2.2% at four different concentrations (5, 25, 50, and 100 μg/L). A simple, faster, and more accurate HPLC-UV method is developed which is very useful for monitoring 5-FC concentration in low volume serum samples without evaporation step and ion exchange chromatography within minutes.

关 键 词：5-FLUOROCYTOSINE Serum Sample HPLC-UV Method Quantification Validation 

分 类 号：R73[医药卫生—肿瘤]