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Development and validation of an ultra-high performance liquid chromatography - high resolution mass spectrometry method for the quantification of total and free teicoplanin in human plasma

Olivier Deltombe (UGent) , Tom Mertens (UGent) , Sunny Eloot (UGent) and Alain Verstraete (UGent)
(2019) CLINICAL BIOCHEMISTRY. 65. p.29-37
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Abstract
Objectives: The antibiotic teicoplanin, used for the treatment of infections caused by Gram-positive bacteria, is highly bound to plasma proteins (percentage protein binding, %PB, around 90%) and therapeutic plasma levels of total teicoplanin are 10-100 mg/L. Because of the low free concentrations (i.e. < 1-10 mg/L), current immunoassays are not able to quantify free teicoplanin concentrations, although they might be more relevant in therapeutic drug monitoring than total concentrations. Design and methods: In this study, an ultra-high performance liquid chromatography - high resolution mass spectrometry (UHPLC-HRMS) method for the quantification of total and free teicoplanin in K(2)EDTA plasma samples was developed and validated. Furthermore, %PB obtained by ultrafiltration was compared with that obtained by equilibrium dialysis using spiked samples from healthy subjects. Analytes were separated using a phenylhexyl column, gradient mobile phase analysis was used, total run time was 4.5 min and teicoplanin was detected by orbitrap MS. Results: The precision and accuracy were below 15% and within +/- 15%, respectively and teicoplanin was found to be stable for at least 14 days in plasma at 4 degrees C. The %PB of teicoplanin in spiked plasma from healthy subjects as obtained by ultrafiltration (94.1 +/- 1.3%) was in good agreement with that obtained by equilibrium dialysis (93.6 +/- 0.4%), whereas mean %PB of teicoplanin in samples from infected patients who received the antibiotic was 87.7 +/- 4.2% (range: 79.6-95.4%). Conclusion: A novel highly sensitive UHPLC-HRMS method was developed and validated for the quantification of total and free teicoplanin in human K(2)EDTA plasma samples. Amongst others, this method is suitable for therapeutic drug monitoring.
Keywords
FLUORESCENCE POLARIZATION IMMUNOASSAY, PROTEIN-BINDING, SERUM, ASSAY, VARIABILITY, VANCOMYCIN, SELECTION, MS, Teicoplanin, Free fraction, Ultrafiltration, Ultra-high performance, liquid chromatography, Orbitrap, High resolution mass spectrometry

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MLA
Deltombe, Olivier et al. “Development and Validation of an Ultra-high Performance Liquid Chromatography - High Resolution Mass Spectrometry Method for the Quantification of Total and Free Teicoplanin in Human Plasma.” CLINICAL BIOCHEMISTRY 65 (2019): 29–37. Print.
APA
Deltombe, O., Mertens, T., Eloot, S., & Verstraete, A. (2019). Development and validation of an ultra-high performance liquid chromatography - high resolution mass spectrometry method for the quantification of total and free teicoplanin in human plasma. CLINICAL BIOCHEMISTRY, 65, 29–37.
Chicago author-date
Deltombe, Olivier, Tom Mertens, Sunny Eloot, and Alain Verstraete. 2019. “Development and Validation of an Ultra-high Performance Liquid Chromatography - High Resolution Mass Spectrometry Method for the Quantification of Total and Free Teicoplanin in Human Plasma.” Clinical Biochemistry 65: 29–37.
Chicago author-date (all authors)
Deltombe, Olivier, Tom Mertens, Sunny Eloot, and Alain Verstraete. 2019. “Development and Validation of an Ultra-high Performance Liquid Chromatography - High Resolution Mass Spectrometry Method for the Quantification of Total and Free Teicoplanin in Human Plasma.” Clinical Biochemistry 65: 29–37.
Vancouver
1.
Deltombe O, Mertens T, Eloot S, Verstraete A. Development and validation of an ultra-high performance liquid chromatography - high resolution mass spectrometry method for the quantification of total and free teicoplanin in human plasma. CLINICAL BIOCHEMISTRY. 2019;65:29–37.
IEEE
[1]
O. Deltombe, T. Mertens, S. Eloot, and A. Verstraete, “Development and validation of an ultra-high performance liquid chromatography - high resolution mass spectrometry method for the quantification of total and free teicoplanin in human plasma,” CLINICAL BIOCHEMISTRY, vol. 65, pp. 29–37, 2019.
@article{8609498,
  abstract     = {Objectives: The antibiotic teicoplanin, used for the treatment of infections caused by Gram-positive bacteria, is highly bound to plasma proteins (percentage protein binding, %PB, around 90%) and therapeutic plasma levels of total teicoplanin are 10-100 mg/L. Because of the low free concentrations (i.e. < 1-10 mg/L), current immunoassays are not able to quantify free teicoplanin concentrations, although they might be more relevant in therapeutic drug monitoring than total concentrations. 
Design and methods: In this study, an ultra-high performance liquid chromatography - high resolution mass spectrometry (UHPLC-HRMS) method for the quantification of total and free teicoplanin in K(2)EDTA plasma samples was developed and validated. Furthermore, %PB obtained by ultrafiltration was compared with that obtained by equilibrium dialysis using spiked samples from healthy subjects. Analytes were separated using a phenylhexyl column, gradient mobile phase analysis was used, total run time was 4.5 min and teicoplanin was detected by orbitrap MS. 
Results: The precision and accuracy were below 15% and within +/- 15%, respectively and teicoplanin was found to be stable for at least 14 days in plasma at 4 degrees C. The %PB of teicoplanin in spiked plasma from healthy subjects as obtained by ultrafiltration (94.1 +/- 1.3%) was in good agreement with that obtained by equilibrium dialysis (93.6 +/- 0.4%), whereas mean %PB of teicoplanin in samples from infected patients who received the antibiotic was 87.7 +/- 4.2% (range: 79.6-95.4%). 
Conclusion: A novel highly sensitive UHPLC-HRMS method was developed and validated for the quantification of total and free teicoplanin in human K(2)EDTA plasma samples. Amongst others, this method is suitable for therapeutic drug monitoring.},
  author       = {Deltombe, Olivier and Mertens, Tom and Eloot, Sunny and Verstraete, Alain},
  issn         = {0009-9120},
  journal      = {CLINICAL BIOCHEMISTRY},
  keywords     = {FLUORESCENCE POLARIZATION IMMUNOASSAY,PROTEIN-BINDING,SERUM,ASSAY,VARIABILITY,VANCOMYCIN,SELECTION,MS,Teicoplanin,Free fraction,Ultrafiltration,Ultra-high performance,liquid chromatography,Orbitrap,High resolution mass spectrometry},
  language     = {eng},
  pages        = {29--37},
  title        = {Development and validation of an ultra-high performance liquid chromatography - high resolution mass spectrometry method for the quantification of total and free teicoplanin in human plasma},
  url          = {http://dx.doi.org/10.1016/j.clinbiochem.2018.12.010},
  volume       = {65},
  year         = {2019},
}

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