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Development and validation of an LC tandem MS assay for the quantification of β-lactam antibiotics in the sputum of cystic fibrosis patients

Katrien Forier (UGent) , Virginie Van Heck, Mieke Carlier (UGent) , Eva Van Braeckel (UGent) , Sabine Van daele (UGent) , Frans De Baets (UGent) , Petra Schelstraete (UGent) , Filomeen Haerynck (UGent) , Veronique Stove (UGent) , Leen Van Simaey (UGent) , et al.
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Abstract
Objectives: Antibiotic therapy is of vital importance for the control of infectious exacerbations in cystic fibrosis (CF) patients. However, very little is known regarding the fraction of systemically administered antibiotics reaching the lower respiratory tract secretions. We developed and validated amethod to measure the concentrations of piperacillin, ceftazidime, meropenem and aztreonamin CF sputum, and present the validation data. Methods: Ultra-performance LC coupled to tandem MS was used. A single sample can be measured in 2.5 min with multiple reaction monitoring in positive electrospray ionization mode. Deuterated internal standards were used and a concentration range of 0.7-160 mg/L was covered. The method was validated according to the EMA guideline on analytical method validation. Results: The boundaries within which a reliable measurement in CF sputum can be performed were determined. A few constraints are linked to the instability of the antibiotics in sputum. Piperacillin showed limited stability at room temperature and during freeze-thaw cycles. Autosampler instability was observed after 15 h for aztreonam at low concentrations. Conclusions: The method allows a reliable measurement of the selected antibiotics, if precautions are taken regarding the limited stability of piperacillin at room temperature. Due to freeze-thaw instability, piperacillin should always be analysed on the day of sampling. Quick review of the analytical data and reanalysis are needed as low concentrations of aztreonam are not stable in the autosampler.
Keywords
PSEUDOMONAS-AERUGINOSA, BRONCHOPULMONARY INFECTION, BIOANALYTICAL METHODS, LUNG INFECTION, HPLC-MS/MS, CARBENICILLIN, PIPERACILLIN, PENETRATION, STRATEGIES, TOBRAMYCIN

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Chicago
Forier, Katrien, Virginie Van Heck, Mieke Carlier, Eva Van Braeckel, Van daeleSabine, Frans De Baets, Petra Schelstraete, et al. 2018. “Development and Validation of an LC Tandem MS Assay for the Quantification of Β-lactam Antibiotics in the Sputum of Cystic Fibrosis Patients.” Journal of Antimicrobial Chemotherapy 73 (1): 95–101.
APA
Forier, K., Van Heck, V., Carlier, M., Van Braeckel, E., Van daeleSabine, De Baets, F., Schelstraete, P., et al. (2018). Development and validation of an LC tandem MS assay for the quantification of β-lactam antibiotics in the sputum of cystic fibrosis patients. JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, 73(1), 95–101.
Vancouver
1.
Forier K, Van Heck V, Carlier M, Van Braeckel E, Van daeleSabine, De Baets F, et al. Development and validation of an LC tandem MS assay for the quantification of β-lactam antibiotics in the sputum of cystic fibrosis patients. JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY. 2018;73(1):95–101.
MLA
Forier, Katrien, Virginie Van Heck, Mieke Carlier, et al. “Development and Validation of an LC Tandem MS Assay for the Quantification of Β-lactam Antibiotics in the Sputum of Cystic Fibrosis Patients.” JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY 73.1 (2018): 95–101. Print.
@article{8540482,
  abstract     = {Objectives: Antibiotic therapy is of vital importance for the control of infectious exacerbations in cystic fibrosis (CF) patients. However, very little is known regarding the fraction of systemically administered antibiotics reaching the lower respiratory tract secretions. We developed and validated amethod to measure the concentrations of piperacillin, ceftazidime, meropenem and aztreonamin CF sputum, and present the validation data. 
Methods: Ultra-performance LC coupled to tandem MS was used. A single sample can be measured in 2.5 min with multiple reaction monitoring in positive electrospray ionization mode. Deuterated internal standards were used and a concentration range of 0.7-160 mg/L was covered. The method was validated according to the EMA guideline on analytical method validation. 
Results: The boundaries within which a reliable measurement in CF sputum can be performed were determined. A few constraints are linked to the instability of the antibiotics in sputum. Piperacillin showed limited stability at room temperature and during freeze-thaw cycles. Autosampler instability was observed after 15 h for aztreonam at low concentrations. 
Conclusions: The method allows a reliable measurement of the selected antibiotics, if precautions are taken regarding the limited stability of piperacillin at room temperature. Due to freeze-thaw instability, piperacillin should always be analysed on the day of sampling. Quick review of the analytical data and reanalysis are needed as low concentrations of aztreonam are not stable in the autosampler.},
  author       = {Forier, Katrien and Van Heck, Virginie and Carlier, Mieke and Van Braeckel, Eva and Van daele, Sabine and De Baets, Frans and Schelstraete, Petra and Haerynck, Filomeen and Stove, Veronique and Van Simaey, Leen and Vaneechoutte, Mario and Verstraete, Alain},
  issn         = {0305-7453},
  journal      = {JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY},
  keyword      = {PSEUDOMONAS-AERUGINOSA,BRONCHOPULMONARY INFECTION,BIOANALYTICAL METHODS,LUNG INFECTION,HPLC-MS/MS,CARBENICILLIN,PIPERACILLIN,PENETRATION,STRATEGIES,TOBRAMYCIN},
  language     = {eng},
  number       = {1},
  pages        = {95--101},
  title        = {Development and validation of an LC tandem MS assay for the quantification of \ensuremath{\beta}-lactam antibiotics in the sputum of cystic fibrosis patients},
  url          = {http://dx.doi.org/10.1093/jac/dkx331},
  volume       = {73},
  year         = {2018},
}

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