TY - JOUR
T1 - Method development for the determination of teicoplanin in patient serum by solid phase extraction and micellar electrokinetic chromatography
AU - Tsai, I. Lin
AU - Wu, Fe Lin Lin
AU - Gau, Churn Shiouh
AU - Kuo, Ching Hua
N1 - Funding Information:
This study was supported by a research grant (NSC 97-2320-B-002-056) from the National Science Council, Taiwan. We would like to thank Dr. Shao-Wen Sun and Dr. Wheyong Lo for editing this manuscript. We also acknowledge Bureau of food and drug analysis, Department of Health for HPLC/MS/MS analysis of teicoplanin components.
PY - 2009/1/15
Y1 - 2009/1/15
N2 - 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 20 mM 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 μg/mL to 55 μg/mL. Limit of detection (LOD) for each of the six components was found to be lower than 0.06 μg/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.
AB - 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 20 mM 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 μg/mL to 55 μg/mL. Limit of detection (LOD) for each of the six components was found to be lower than 0.06 μg/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.
KW - Micellar electrokinetic chromatography
KW - Serum
KW - Solid phase extraction
KW - Teicoplanin
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U2 - 10.1016/j.talanta.2008.08.022
DO - 10.1016/j.talanta.2008.08.022
M3 - Article
C2 - 19064114
AN - SCOPUS:57049126503
VL - 77
SP - 1208
EP - 1216
JO - Talanta
JF - Talanta
SN - 0039-9140
IS - 3
ER -