Limits...
Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

View Article: PubMed Central - PubMed

ABSTRACT

As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 µg/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (ρ, 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment.

No MeSH data available.


Related in: MedlinePlus

Passing-Bablok regression with regression equations, Pearson's rho, significance levels and Bland-Altman plots between measurements in DBS and plasma for second-line anti-TB drugs: kanamycin, cycloserine, moxifloxacin, levofloxacin, prothionamide, PAS, linezolid, and clarithromycin.Abbreviations: DBS, dried blood spot; PAS, para-aminosalicylic acid.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4940494&req=5

Figure 1: Passing-Bablok regression with regression equations, Pearson's rho, significance levels and Bland-Altman plots between measurements in DBS and plasma for second-line anti-TB drugs: kanamycin, cycloserine, moxifloxacin, levofloxacin, prothionamide, PAS, linezolid, and clarithromycin.Abbreviations: DBS, dried blood spot; PAS, para-aminosalicylic acid.

Mentions: Plasma and DBSs for the comparison study were obtained from residual samples of TB patients. This research was approved by the Seoul National University Bundang Hospital Institutional Review Board, and was conducted in accordance with the Declaration of Helsinki. Passing-Bablok regression analysis revealed strong similarity between drug concentrations in DBSs and in plasma (Fig. 1). For streptomycin, only two residual samples were tested, and the results were 4.8/3.6 µg/mL and 2.1/2.7 µg/mL (plasma/DBS) respectively. The Pearson's rho (ρ) of drugs ranged from 0.798 to 0.989 (P<0.0015). Although these values were lower than those of published single or dual drug assays [789], they are acceptable for TDM in MDR/XDR management. The concentration of each drug in DBSs showed a degree of negative bias compared with that in plasma, except for levofloxacin. Several factors cause a mismatch of drug concentration between plasma and venous whole blood, including differential partition between red blood cells and plasma, and individual hematocrit variations [6]. Nevertheless, a very high positive correlation for clarithromycin, cycloserine, and PAS and high positive correlation for the other drugs were observed [15]. At first, the corresponding plasma values could be calculated from the DBSs by using the conversion equations for clarithromycin, cycloserine and PAS.


Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry
Passing-Bablok regression with regression equations, Pearson's rho, significance levels and Bland-Altman plots between measurements in DBS and plasma for second-line anti-TB drugs: kanamycin, cycloserine, moxifloxacin, levofloxacin, prothionamide, PAS, linezolid, and clarithromycin.Abbreviations: DBS, dried blood spot; PAS, para-aminosalicylic acid.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940494&req=5

Figure 1: Passing-Bablok regression with regression equations, Pearson's rho, significance levels and Bland-Altman plots between measurements in DBS and plasma for second-line anti-TB drugs: kanamycin, cycloserine, moxifloxacin, levofloxacin, prothionamide, PAS, linezolid, and clarithromycin.Abbreviations: DBS, dried blood spot; PAS, para-aminosalicylic acid.
Mentions: Plasma and DBSs for the comparison study were obtained from residual samples of TB patients. This research was approved by the Seoul National University Bundang Hospital Institutional Review Board, and was conducted in accordance with the Declaration of Helsinki. Passing-Bablok regression analysis revealed strong similarity between drug concentrations in DBSs and in plasma (Fig. 1). For streptomycin, only two residual samples were tested, and the results were 4.8/3.6 µg/mL and 2.1/2.7 µg/mL (plasma/DBS) respectively. The Pearson's rho (ρ) of drugs ranged from 0.798 to 0.989 (P<0.0015). Although these values were lower than those of published single or dual drug assays [789], they are acceptable for TDM in MDR/XDR management. The concentration of each drug in DBSs showed a degree of negative bias compared with that in plasma, except for levofloxacin. Several factors cause a mismatch of drug concentration between plasma and venous whole blood, including differential partition between red blood cells and plasma, and individual hematocrit variations [6]. Nevertheless, a very high positive correlation for clarithromycin, cycloserine, and PAS and high positive correlation for the other drugs were observed [15]. At first, the corresponding plasma values could be calculated from the DBSs by using the conversion equations for clarithromycin, cycloserine and PAS.

View Article: PubMed Central - PubMed

ABSTRACT

As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 &micro;g/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (&rho;, 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment.

No MeSH data available.


Related in: MedlinePlus