High Potential Agricultural Growth Areas: Alternative Approaches and Considerations
Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 ·...
Transcript of Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 ·...
![Page 1: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/1.jpg)
Approaches and technologies offering potential solutions to the problems associated with DBS analyses
Neil Spooner Ph.D., C.Chem, FRSC
Founder & DirectorSpooner Bioanalytical Solutions Ltd
Senior Visiting Research FellowSchool of Life and Medical Sciences, University of Hertfordshire
NEIL SPOONER - ISNS 2016 1
![Page 2: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/2.jpg)
Effects of Blood Hematocrit Has an effect on the diameter of derived DBS sample, resulting in unknown volume in sub-punch of sample
May have an effect on the recovery of analyte
Result is reduced (unknown) analytical accuracy
NEIL SPOONER - ISNS 2016 2
70%45%20%
![Page 3: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/3.jpg)
Alternative Approaches for Collecting Dried Blood SamplesVariety of technologies aimed at collecting fixed volume of blood regardless of hematocrit, resulting in improved accuracy of determined concentrations
NEIL SPOONER - ISNS 2016 3
![Page 4: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/4.jpg)
Volumetric Absorptive Microsampling - Mitra
Hydrophilic porous material
Each Tip has a fixed, highly reproducible internal porous volume - 10 µL
◦ Regardless of blood hematocrit
Rapid wicking◦ Under 6 seconds)
NEIL SPOONER - ISNS 2016 4
![Page 5: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/5.jpg)
Simple workflow
NEIL SPOONER - ISNS 2016 5
![Page 6: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/6.jpg)
Simple to use
NEIL SPOONER - ISNS 2016 6
![Page 7: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/7.jpg)
Mitra volumetric sampling performance
NEIL SPOONER - ISNS 2016 7
Denniff & Spooner (2014) Anal. Chem. 86, 8489-8495, Denniff et al (2015) J. Pharm. Biomed. Anal. 108, 61-69,
Spooner et al (2015) Bioanalysis 7(6), 653-659
• Human blood at different HCTs was spiked with 14C caffeine
• Tip oxidised to 14CO2
• Radioactivity determined by scintillation counting
![Page 8: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/8.jpg)
hemaPEN™ Workflow
NEIL SPOONER - ISNS 2016 8
For Research Purposes only - this technology has not been approved by the regulatory authorities
![Page 9: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/9.jpg)
hemaPEN™ Construction
NEIL SPOONER - ISNS 2016 9
For Research Purposes only - this technology has not been approved by the regulatory authorities
![Page 10: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/10.jpg)
hemaPEN™ Volumetric Performance
NEIL SPOONER - ISNS 2016 10
For Research Purposes only - this technology has not been approved by the regulatory authorities
Human blood at different
HCTs spiked with caffeine
(5mg/mL)
3 µL blood spotted onto 226
substrate using;
o Air displacement pipette
o Positive displacement
pipette
o Glass capillary
Aqueous extracts analysed by
HPLC UV/Vis (276 nm)
![Page 11: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/11.jpg)
NEIL SPOONER - ISNS 2016 11
Leuthold et al. (2015), Anal.
Chem. 87, 2068-2071
![Page 12: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/12.jpg)
CapiTainer AB
NEIL SPOONER - ISNS 2016 12
Activation
Channel filling
Inlet dissolving
Outlet dissolving
Outlet activation
Punch out
Lenk et al (2015)
Bioanalysis 7(16),
2085-2094.
![Page 13: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/13.jpg)
…..and there’s more
NEIL SPOONER - ISNS 2016 13
![Page 14: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/14.jpg)
Approaches for Normalising Quantitative Data Obtained by Conventional DBS SamplingVariety of approaches aimed at correcting the measured concentration
NEIL SPOONER - ISNS 2016 14
![Page 15: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/15.jpg)
Add internal standard to DBS sample prior to extraction
NEIL SPOONER - ISNS 2016 15
Abu-Rabie et al. (2015) Anal. Chem. 87, 4996-5003
![Page 16: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/16.jpg)
Normalise data to hematocrit K+ determination in aqueous extract of 3-mm DBS punch using a routine clinical chemistry analyzer
•[K+] intracellularly >> [K+] plasma and > 99% of cells are RBCs
•[K+] are tightly controlled � no large interindividual variability
•K+ is universal
•K+ proved to be stable in DBS
•K+ easily measurable in DBS
NEIL SPOONER - ISNS 2016. 16
Capiau et al. (2013) Anal. Chem. 85, 404-410; De Kesel et al. (2014), Anal. Bioanal. Chem. 406, 6749
Raw Data
Corrected for HCT
![Page 17: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/17.jpg)
Normalise data to hematocrit
NEIL SPOONER - ISNS 2016 17
Capiau et al. (2016) Anal. Chem. 88, 6538-6546
Non-contact diffuse reflectance spectroscopy to
determine different hemoglobin forms
![Page 18: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/18.jpg)
Normalise data to an endogenous component
NEIL SPOONER - ISNS 2016 18
![Page 19: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/19.jpg)
Determination of blood volume by conductance
Electrical conductivity measured by ring-disk electrode in 100 µL aqueous extracts of spotted blood
NEIL SPOONER - ISNS 2016 19
![Page 20: Approaches and technologies offering potential solutions to the … Spooner... · 2016-09-22 · Approaches and technologies offering potential solutions to the problems associated](https://reader035.fdocuments.in/reader035/viewer/2022081611/5f0a12017e708231d429df8c/html5/thumbnails/20.jpg)
Points to Consider Quality/accuracy of analytical data is dependant on the quality of the sample collected
◦ Blood hematocrit introduces a bias into quantitative data based upon lack of control of blood volume and recovery with conventional DBS sample collection approaches
Consider use of alternative blood collection approaches if;
◦ Determined data is to be compared between labs, or across time
◦ The differences between measured values for a positive and negative result are marginal
Can alternative technologies and workflows be introduced to further enable automation? Consider potential for;
◦ Increased data quality
◦ Cost savings
NEIL SPOONER - ISNS 2016 20