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What is the Hematocrit effect ?
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New Ht independent dried blood spot cards discussion Ynze Mengerink - - PowerPoint PPT Presentation
New Ht independent dried blood spot cards discussion Ynze Mengerink - - PowerPoint PPT Presentation
New Ht independent dried blood spot cards discussion Ynze Mengerink Barcelona, November 21 st 2013 What is the Hematocrit effect ? Page 1 DBS Card 2 drops 2 drops 2 drops 1 drops 2 drops 3 drops Ht 20 Ht 45 Ht 80 Ht 45
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DBS Card
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Punch center Each Punch contains same volume of blood Punch center 2 drops 2 drops 2 drops Ht 20 Ht 45 Ht 80 1 drops 2 drops 3 drops Ht 45 Ht 45 Ht 45 Each Punch contains not the same volume of blood
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Ht effect on commercial available DBS Cards
3 0,75 0,80 0,85 0,90 0,95 1,00 1,05 1,10 1,15 1,20 1,25 20 30 40 50 60 70 80 Normalized spot size (Ht45% =1) % Hematocrit
Influence of Hematocrit on Spot Size
903 DMPK B DMPK A Bond Elute DMS DMPK C
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What causes the Hematocrit effect ?
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Page 19 0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 1 2 3 4 5 6 7 8 diameter of spot (arbitrary units) time of spot formation (arbitrary units)
Bloodspot spreading
Whatman S pot size Whatman Core
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How does our new material work
- Woven polyester material, fiber and mesh size in µm range
– Large enough to transfer blood cells and small enough to retain dried blood
- Multi layers
– Typically eight layers, spot size relates to number of layers
- Coated with a very hydrophilic coating and UV cured
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After a few seconds
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After a few minutes
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Page 28 0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 1 2 3 4 5 6 7 8 diameter of spot (arbitrary units) time of spot formation (arbitrary units)
Bloodspot spreading
DS M card S pot size DS M card Core
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Page 29 0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 1 2 3 4 5 6 7 8 diameter of spot (arbitrary units) time of spot formation (arbitrary units)
Bloodspot spreading
DS M card S pot size DS M card Core Whatman S pot size Whatman Core
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Influence of Hematocrit
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So, this looks like a holy grail to reduce study animals ??
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4 layers no orientation of layers
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After a few seconds
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After a few minutes
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After an hour
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S ingle layer – aged blood
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0 graden
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3 Degrees angle
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10 graden
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15 graden
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45 graden
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Orientation of layers
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How the card is build
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Not orientated material
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Orientated material
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Results: Spot Homogeneity
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LCMSMS Card 1 Card 2 PUNCH SPOT 1 SPOT 2 SPOT 3 SPOT 4 SPOT 5 SPOT 6 AVG RSD MIDDLE
- 10. 1
- 9. 7
- 9. 8
- 9. 8
- 10. 3
- 10. 4
- 10. 0
3 LEFT
- 10. 4
- 9. 9
- 10. 1
- 10. 3
- 10. 2
- 9. 4
- 10. 1
3 RIGHT
- 10. 8
- 10. 7
- 10. 4
- 11. 7
- 11. 5
- 10. 7
- 11. 0
5 DOWN
- 11. 1
- 11. 0
- 11. 7
- 10. 1
- 9. 9
- 11. 1
- 10. 8
7 AVG
- 10. 6
- 10. 3
- 10. 5
- 10. 5
- 10. 4
- 10. 4
Average
- 10. 47
RSD 4 6 8 8 7 7 RSD (n=24) 6
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Results: Recovery from full spot
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Results: Dry time
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Results: Tunable spot size
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Results: External tests
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Test at GS K confirmed excellent spot size independency towards Hematocrit, but some preliminary results of tests between whole and punched spot show biased results and are under investigation.
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Conclusions
- Very interesting new material, shows spot sizes, which are
independent of hematocrit value of the blood sample
- Fixed spot size at exact volume, as a consequence, blood
volume on card can be determined on spot size.
- Two patents in place (card and coating)
- A startup company is being established, to commercialize
this product
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Acknowledgements
- DS
M: Jun Qiu (our 007), Ellen Gorissen, Nik de Bont and John Mommers
- Chemelot: Odile S
teij ger, Henri Theunissen and Bert Kip
- Ministry of Economic Affairs
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