Is the patient taking their 'heart' pills? A dried blood spot sample - LC-HRMS assay




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Peer reviewed


Introduction: Whilst cardiovascular disease is the biggest killer in the UK some 60% of patients are reported not to take their medication. The use of blood spot collection cards was investigated as a means of obtaining small volume samples for the analysis of therapeutic drugs for assessing medication adherence in cardiovascular therapy. An accurate mass liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed and validated for the determination of the top three prescribed cardiovascular drugs in the UK, bisoprolol, ramipril and simvastatin, in dried blood spots (DBS). Methods: For the preparation of DBS samples whole blood spiked with the three target analytes was used to produce 30µl blood spots on specimen collection cards. An 8mm disc was cut from the dried blood spot and extracted using methanol: water (70:30, v/v) containing the internal standard, atenolol. Extracts were vortexed, sonicated and then centrifuged. Analysis was performed using gradient chromatographic elution and a mobile phase flow rate of 0.6ml/min and the column oven temperature at 40°C with a run time of 3 min. MS detection was carried out in electrospray positive ion mode at accurate mass m/z 326.2326 for bisoprolol, m/z 417.2384 for ramipril, m/z 441.2611 for simvastatin and m/z 276.1703 for the IS. The developed bioanalytical method was applied to blood spots samples taken from adult volunteers previously administered one or more of the target drugs. Results: Drug recoveries from spiked blood spots were ≥92% for bisoprolol and ramipril and ~43% for simvastatin and the drugs were stable in DBS for at least 12 weeks. Validation of the LC-HRMS method showed good linearity and the accuracy and precision values were within the pre-defined limits of ≤15% at all concentrations. Factors with potential to affect drug quantification measurements such as the matrix effects and volume of blood applied onto the collection card were investigated. Bisoprolol Ramipril Simvastatin
LoQ (S/N >10) 0.5 ng/ml 1.0 ng/ml 5.0 ng/ml
Conclusion: The accurate mass LC-HRMS DBS based method successfully identified control volunteers who were known to be either adherent or non-adherent. There were no false positives from volunteers taking other cardiovascular drugs or from volunteers receiving no medication. Requiring only a micro volume blood sample for analysis, the DBS based assay has the potential to assess patient adherence for a range of therapeutic drugs.



Blood spot analysis, adherence, cardiovascular therapy


Tanna, S., Cocks, E. and Lawson, G. (2013) Is the patient taking their 'heart' pills? A dried blood spot sample - LC-HRMS assay. Proceedings of The 24th International Symposium on Pharmaceutical and Biomedical Analysis, 30 June-3 July 2013


Research Institute

Leicester Institute for Pharmaceutical Innovation - From Molecules to Practice (LIPI)