Critical issues in quantitative bioanalysis of Dried Blood Spot samples
Апстракт
When using DBS as sample collection tool, several specific factors can contribute to assay bias and compromise regulatory-based acceptance criteria. Certain factors like hematocrit (Hct) level, pipettes, sample volume and laboratory personnel have a potential to contribute bioanalytical method bias inherently. The proper understanding of Hct effect and the efficient resolving of the related concerns may determine the future of DBS sampling practice in regulated bioanalysis. Therefore, we aimed at reaching a procedure that enables accurate and precise blood spotting onto the filter paper by simultaneous investigation of factors that were suggested to be scientifically relevant in this context. The effects of five qualitative factors - temperature of blood samples, type of pipettes, pipetting technique, age of blood samples and analyst - were investigated using a multilevel categorical D-optimal design. Five responses were observed in the study (RSD22%Hct, RSD30%Hct, RSD39%Hct, RSD51%Hct..., RSD62%Hct) as they can provide information on the influence of factor settings on the consistency of DBS areas. DBS cards with four spot replicates, corresponding to particular combination of investigated factors defined by experimental plan, were scanned and the area of blood spots was determined by image processing. The principle of backward elimination was applied in computation of the adequate qualitative linear mathematical models with added appropriate two-factor interactions to relate selected responses with studied factors. It was concluded that %RSD value of DBS, regardless Hct levels, is completely independent of type of pipettes and age of blood samples, but can significantly be affected by blood sample temperature, pipetting technique and level of training of analyst. Consequently, the procedure for precise and accurate formation of DBS of uniform area, regardless the Hct value, implies samples at body/room temperature, reversed pipetting technique for rigorous delivery of a sample volume onto the card and a properly trained analyst for handling blood samples. The adequacy of the suggested procedure was confirmed by a verification experiment. Identification of the factors affecting the consistency of DBS formation provided the evidence that this contribution to total assay bias can be successfully controlled and reduced.
Извор:
IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care”, 2023, 48-48Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200161 (Универзитет у Београду, Фармацеутски факултет) (RS-MESTD-inst-2020-200161)
Напомена:
- 4th Poznań Conference "Modern pharmaceutical and biomedical analytics in health care", From 23rd to 24th October 2023, Poznań, Poland, Abstract Book.
Институција/група
PharmacyTY - CONF AU - Malenović, Anđelija AU - Rmandić, Milena AU - Dotsikas, Yannis PY - 2023 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5554 AB - When using DBS as sample collection tool, several specific factors can contribute to assay bias and compromise regulatory-based acceptance criteria. Certain factors like hematocrit (Hct) level, pipettes, sample volume and laboratory personnel have a potential to contribute bioanalytical method bias inherently. The proper understanding of Hct effect and the efficient resolving of the related concerns may determine the future of DBS sampling practice in regulated bioanalysis. Therefore, we aimed at reaching a procedure that enables accurate and precise blood spotting onto the filter paper by simultaneous investigation of factors that were suggested to be scientifically relevant in this context. The effects of five qualitative factors - temperature of blood samples, type of pipettes, pipetting technique, age of blood samples and analyst - were investigated using a multilevel categorical D-optimal design. Five responses were observed in the study (RSD22%Hct, RSD30%Hct, RSD39%Hct, RSD51%Hct, RSD62%Hct) as they can provide information on the influence of factor settings on the consistency of DBS areas. DBS cards with four spot replicates, corresponding to particular combination of investigated factors defined by experimental plan, were scanned and the area of blood spots was determined by image processing. The principle of backward elimination was applied in computation of the adequate qualitative linear mathematical models with added appropriate two-factor interactions to relate selected responses with studied factors. It was concluded that %RSD value of DBS, regardless Hct levels, is completely independent of type of pipettes and age of blood samples, but can significantly be affected by blood sample temperature, pipetting technique and level of training of analyst. Consequently, the procedure for precise and accurate formation of DBS of uniform area, regardless the Hct value, implies samples at body/room temperature, reversed pipetting technique for rigorous delivery of a sample volume onto the card and a properly trained analyst for handling blood samples. The adequacy of the suggested procedure was confirmed by a verification experiment. Identification of the factors affecting the consistency of DBS formation provided the evidence that this contribution to total assay bias can be successfully controlled and reduced. C3 - IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care” T1 - Critical issues in quantitative bioanalysis of Dried Blood Spot samples SP - 48 EP - 48 UR - https://hdl.handle.net/21.15107/rcub_farfar_5554 ER -
@conference{ author = "Malenović, Anđelija and Rmandić, Milena and Dotsikas, Yannis", year = "2023", abstract = "When using DBS as sample collection tool, several specific factors can contribute to assay bias and compromise regulatory-based acceptance criteria. Certain factors like hematocrit (Hct) level, pipettes, sample volume and laboratory personnel have a potential to contribute bioanalytical method bias inherently. The proper understanding of Hct effect and the efficient resolving of the related concerns may determine the future of DBS sampling practice in regulated bioanalysis. Therefore, we aimed at reaching a procedure that enables accurate and precise blood spotting onto the filter paper by simultaneous investigation of factors that were suggested to be scientifically relevant in this context. The effects of five qualitative factors - temperature of blood samples, type of pipettes, pipetting technique, age of blood samples and analyst - were investigated using a multilevel categorical D-optimal design. Five responses were observed in the study (RSD22%Hct, RSD30%Hct, RSD39%Hct, RSD51%Hct, RSD62%Hct) as they can provide information on the influence of factor settings on the consistency of DBS areas. DBS cards with four spot replicates, corresponding to particular combination of investigated factors defined by experimental plan, were scanned and the area of blood spots was determined by image processing. The principle of backward elimination was applied in computation of the adequate qualitative linear mathematical models with added appropriate two-factor interactions to relate selected responses with studied factors. It was concluded that %RSD value of DBS, regardless Hct levels, is completely independent of type of pipettes and age of blood samples, but can significantly be affected by blood sample temperature, pipetting technique and level of training of analyst. Consequently, the procedure for precise and accurate formation of DBS of uniform area, regardless the Hct value, implies samples at body/room temperature, reversed pipetting technique for rigorous delivery of a sample volume onto the card and a properly trained analyst for handling blood samples. The adequacy of the suggested procedure was confirmed by a verification experiment. Identification of the factors affecting the consistency of DBS formation provided the evidence that this contribution to total assay bias can be successfully controlled and reduced.", journal = "IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care”", title = "Critical issues in quantitative bioanalysis of Dried Blood Spot samples", pages = "48-48", url = "https://hdl.handle.net/21.15107/rcub_farfar_5554" }
Malenović, A., Rmandić, M.,& Dotsikas, Y.. (2023). Critical issues in quantitative bioanalysis of Dried Blood Spot samples. in IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care”, 48-48. https://hdl.handle.net/21.15107/rcub_farfar_5554
Malenović A, Rmandić M, Dotsikas Y. Critical issues in quantitative bioanalysis of Dried Blood Spot samples. in IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care”. 2023;:48-48. https://hdl.handle.net/21.15107/rcub_farfar_5554 .
Malenović, Anđelija, Rmandić, Milena, Dotsikas, Yannis, "Critical issues in quantitative bioanalysis of Dried Blood Spot samples" in IV Poznańska Konferencja Naukowo – Szkoleniowej - „Modern pharmaceutical and biomedical analytics in health care” (2023):48-48, https://hdl.handle.net/21.15107/rcub_farfar_5554 .