CHAPTER 5 SUMMARY AND PERSPECTIVE
5.1 Summary and Perspective
In this dissertation we developed several methods to improve DBS sampling technique applications in pharmaceutical and metabolomics clinical applications. Our approaches mainly focused on two major issues: 1) improvement of DBS assay sensitivity by improving MS ionization efficiency (chapter 2) and 2) development of PCI-IS assisted LC-ESI-MS/MS methods to resolve the effects of spotted blood volume for monitoring of anti-HIV drugs and for DBS-based metabolomics studies (Chapter 3 and 4).
Improvement of DBS assay sensitivity is still required to monitor the low therapeutic concentrations using LC-ESI-MS/MS. In order to achieve the sensitive detection for abused drugs, we incorporated the IB source, which greatly improved the method sensitivity when compared to the ESI source, and rendered the proposed method applicability for the detection of trace amounts of abused drugs on the DBS card. The reported method provided a new direction for abused drug screening using DBS.
In order to increase the applicability of the DBS sampling technique in drug monitoring, we developed a PCI-IS method to estimate the blood volume on DBS cards and to quantify 6 anti-HIV drugs in DBS samples in chapter 3. We successfully applied our developed method to quantify AR drugs in paired DBS and plasma samples and correlated their concentrations using conversion factors. Bland-Altman plot revealed that predicted plasma concentrations from DBS concentrations showed good agreements with the measured plasma concentrations after translated using CFs. Newly approved anti-HIV drugs added in the developed method could facilitate a reliable drug quantification method for clinical trials, pharmacokinetic studies, and also for TDM when small sample volumes are available. The method we described is able to support the investigation of the efficacy of cART in rural areas with patient self-sampling on DBS cards. We anticipate
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that the developed method could increase the applicability of the DBS sampling technique in clinical monitoring.
PCI-IS assisted LC-ESI-MS/MS analysis developed for DBS-based metabolomics studies in chapter 4 simultaneously calibrated matrix effects which enabled the evaluation of the effects of spot volumes and Hct on target metabolites, and estimated the blood volume on DBS cards. The whole spot analysis combined with PCI-IS strategy provided a solution to the challenging problems associated with DBS sampling techniques.
The spot volume estimation on DBS cards using PCI-IS strategy could facilitate the volume correction procedure for accurate quantitative DBS-based metabolomics studies.
Investigation of Hct variation effect revealed that it affected endogenous metabolite concentrations in DBS samples. Thus, pre-investigation of Hct variation effect for metabolite distribution is necessary, and concentration normalization is important based on their distribution in blood for quantitative DBS-based metabolomics studies. This study demonstrated the ability of the PCI-IS method to simultaneously estimate blood volume, calibrate ion source contaminated signal change, and matrix effect, which could effectively improve analytical performance of DBS-base metabolomics. Through resolving critical problems of DBS-based metabolomics, DBS-based metabolomics could be applied to more clinical research applications.
In conclusion, we demonstrated that PCI-IS assisted LC-ESI-MS/MS analytical methods could solve the challenges associate with the DBS sampling technique and it could increase the applicability of DBS in clinical practice. We anticipate these developed methods to improve the accuracy of drug monitoring and improve data integrity for various DBS-based metabolomics studies and benefit precision medicine.
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Chapter 6
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