Method development for the determination of teicoplanin in patient serum by solid phase extraction and micellar electrokinetic chromatography.
In-hospital deaths caused by the infection of methicillin-resistant Staphylococcus aureus (MRSA) are on the increase worldwide. Teicoplanin is a potent glycopeptide antibiotic against MRSA. A rapid and cost-saving micellar electrokinetic chromatography (MEKC) method combined with solid phase extraction (SPE) was developed and then validated to quantify teicoplanin in patient serum in this work. The method includes the following steps: (1) pretreatment of the serum samples with 10 M urea to denature proteins, (2) application of SPE by using an OASIS HLB cartridge to clean up and concentrate the serum samples, and (3) use of MEKC for sample analysis. Under the optimized conditions, the SPE recovery of teicoplanin is higher than 90%. The six major components of teicoplanin could be baseline-separated from one another and endogenous materials in 12 min with a background electrolyte composed of 20mM sodium tetraborate buffer pH 8.8, 40 mM sodium dodecyl sulfate, and 11% (v/v) ACN. The relative standard deviation (R.S.D.) of the peak area ratios for method repeatability (n=6) and intermediate precision (inter-day, n=3) were found to be lower than 4.18% and 5.30%, respectively. The calibration curves were linear between the chromatographic response and total teicoplanin concentration over the range of 5 microg/mL to 55 microg/mL. Limit of detection (LOD) for each of the six components was found to be lower than 0.06 microg/mL. Pearson's correlation revealed that a good correlation (r=0.98) was obtained between the SPE-MEKC method and the fluorescence polarization immunoassay (FPIA) method. The developed method can be used to quantitatively determine serum teicoplanin concentration in patients for dose monitoring and clinical research.
Tsai
I-Lin
IL
School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan.
Wu
Fe-Lin Lin
FL
Gau
Churn-Shiouh
CS
Kuo
Ching-Hua
CH
D002021:Buffers; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D004563:Electrochemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007700:Kinetics; D015394:Molecular Structure; D012967:Sodium Dodecyl Sulfate; D052616:Solid Phase Extraction / Q000379:methods*; D017334:Teicoplanin / Q000097:blood* / Q000737:chemistry*
D002021:Buffers; D012967:Sodium Dodecyl Sulfate; D017334:Teicoplanin