TY - JOUR
T1 - An on-spot internal standard addition approach for accurately determining colistin A and colistin B in dried blood spots using ultra high-performance liquid chromatography–tandem mass spectrometry
AU - Tsai, I. Lin
AU - Kuo, Ching Hua
AU - Sun, Hsin Yun
AU - Chuang, Yu Chung
AU - Chepyala, Divyabharathi
AU - Lin, Shu Wen
AU - Tsai, Yun Jung
N1 - Funding Information:
This study was supported by the Taipei Medical University ( TMU104-AE1-B21 ). We also thank the NTU Integrated Core Facility for Functional Genomics of the National Research Program for Genomic Medicine of Taiwan for the technical assistance.
Publisher Copyright:
© 2017 Elsevier B.V.
PY - 2017/10/25
Y1 - 2017/10/25
N2 - Outbreaks of multidrug-resistant Gram-negative bacterial infections have been reported worldwide. Colistin, an antibiotic with known nephrotoxicity and neurotoxicity, is now being used to treat multidrug-resistant Gram-negative strains. In this study, we applied an on-spot internal standard addition approach coupled with an ultra high-performance liquid chromatography-tandem mass spectrometry (LC–MS/MS) method to quantify colistin A and B from dried blood spots (DBSs). Only 15 μL of whole blood was required for each sample. An internal standard with the same yield of extraction recoveries as colistin was added to the spot before sample extraction for accurate quantification. Formic acid in water (0.15%) with an equal volume of acetonitrile (50:50 v/v) was used as the extraction solution. With the optimized extraction process and LC–MS/MS conditions, colistin A and B could be quantified from a DBS with respective limits of quantification of 0.13 and 0.27 μg mL−1, and the retention times were < 2 min. The relative standard deviations of within-run and between-run precisions for peak area ratios were all < 17.3%. Accuracies were 91.5-111.2% for lower limit of quantification, low, medium, and high QC samples. The stability of the easily hydrolyzed prodrug, colistin methanesulfonate, was investigated in DBSs. Less than 4% of the prodrug was found to be hydrolyzed in DBSs at room temperature after 48 h. The developed method applied an on-spot internal standard addition approach which benefited the precision and accuracy. Results showed that DBS sampling coupled with the sensitive LC–MS/MS method has the potential to be an alternative approach for colistin quantification, where the bias of prodrug hydrolysis in liquid samples is decreased.
AB - Outbreaks of multidrug-resistant Gram-negative bacterial infections have been reported worldwide. Colistin, an antibiotic with known nephrotoxicity and neurotoxicity, is now being used to treat multidrug-resistant Gram-negative strains. In this study, we applied an on-spot internal standard addition approach coupled with an ultra high-performance liquid chromatography-tandem mass spectrometry (LC–MS/MS) method to quantify colistin A and B from dried blood spots (DBSs). Only 15 μL of whole blood was required for each sample. An internal standard with the same yield of extraction recoveries as colistin was added to the spot before sample extraction for accurate quantification. Formic acid in water (0.15%) with an equal volume of acetonitrile (50:50 v/v) was used as the extraction solution. With the optimized extraction process and LC–MS/MS conditions, colistin A and B could be quantified from a DBS with respective limits of quantification of 0.13 and 0.27 μg mL−1, and the retention times were < 2 min. The relative standard deviations of within-run and between-run precisions for peak area ratios were all < 17.3%. Accuracies were 91.5-111.2% for lower limit of quantification, low, medium, and high QC samples. The stability of the easily hydrolyzed prodrug, colistin methanesulfonate, was investigated in DBSs. Less than 4% of the prodrug was found to be hydrolyzed in DBSs at room temperature after 48 h. The developed method applied an on-spot internal standard addition approach which benefited the precision and accuracy. Results showed that DBS sampling coupled with the sensitive LC–MS/MS method has the potential to be an alternative approach for colistin quantification, where the bias of prodrug hydrolysis in liquid samples is decreased.
KW - Colistin
KW - Dried blood spot
KW - Mass spectrometry
KW - On-spot internal standard addition
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U2 - 10.1016/j.jpba.2017.08.002
DO - 10.1016/j.jpba.2017.08.002
M3 - Article
C2 - 28818804
AN - SCOPUS:85029294366
SN - 0731-7085
VL - 145
SP - 783
EP - 793
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
ER -