SDifLSpray Drying for Long py yg g -TSt fRdBldCllTerm ... · objective is to explore spray drying...
1
S D i f L T St f R d Bl d C ll Spray Drying for Long Term Storage of Red Blood Cells Spray Drying for Long-Term Storage of Red Blood Cells Spray Drying for Long-Term Storage of Red Blood Cells Spray Drying for Long Term Storage of Red Blood Cells Meehe Kim Mary McLean John Simeles Dr Adam Higgins (Sponsor) Meehe Kim, Mary McLean, John Simeles, Dr. Adam Higgins (Sponsor) Meehe Kim, Mary McLean, John Simeles, Dr. Adam Higgins (Sponsor) School of Chemical Biological and Environmental Engineering School of Chemical Biological and Environmental Engineering http://en.wikipedia.org/wiki/File:Erythrocyte_deoxy.jpg1 School of Chemical, Biological, and Environmental Engineering M th d S D T tR lt Obj ti Methods Spray Dry Test Results Objective Methods Spray Dry Test Results Objective R d bl d ll d tdf t f i i ft 30 d i th fi t T t tk t diff t iti th t d i t ti A 80 °C Red blood cells donated for transfusion expire after 30 days in the refrigerator. R d Bl d C ll S i Temperature was taken at different points in the system during testing. An 80 °C Storing and transporting refrigerated blood is costly and inconvenient The Red Blood Cell Suspension l t tt t di t d f t t b it k t Storing and transporting refrigerated blood is costly and inconvenient. The nozzle output temperature was used instead of room temperature because it kept objective is to explore spray drying red blood cells as a means of storing cells at ll i t t hil till i i th t f D i ffi i t tdb objective is to explore spray drying red blood cells as a means of storing cells at cells intact, while still increasing the mass transfer. Drying efficiency was tested by room temperature to improve the logistics of storage i fl t d li t ti (Fi 1) R dbl d ll room temperature to improve the logistics of storage. Slide Test varying gas flow rates and saline concentrations (Figure 1). Red blood cells were Slide Test d th h th l ith h idifi d it t diff ti t th d th d sprayed through the nozzle with humidified nitrogen to differentiate the death due t i f th d th d t d i H l bi i l d h d bl d B k d to spraying from the death due to drying. Hemoglobin is released when red blood Background ll l d b tifi d b t ht t (Fi 2) Th b b Background cells lyse and can be quantified by spectrophotometry (Figure 2). The absorbance Red blood cells are contained by fragile phospholipid membranes and can lyse or fold in Ob i bl d Dilute in phosphate C t if t f td dd tdh l bi i di t d th t 430 i th Red blood cells are contained by fragile phospholipid membranes and can lyse or fold in di i h bl Obtain blood buffered saline Centrifuge spectrum of oxygenated and deoxygenated hemoglobin indicated that 430 nm is the conditions shown below. buffered saline Extract b t l th t tk tf b th C lld th t it fl t f Extract best wavelength to take measurements for both. Cell death at nitrogen flow rates of 10 L/ i i ifi tl l th t 20 L/ i dl f bl d fl t Spray Test 10 L/min are significantly lower than at 20 L/min regardless of blood flow rate. E lt E t Spray Test Excess salt Excess water Temperature < 20 °C Temperature > 80 °C Red Blood Cell Viability Testing Slow drying time Fast drying time Red Blood Cell Viability Testing Slow drying time Fast drying time http://www.chrisrepetsky.com/?m=200910 An external gas atomizer nozzle uses a high-velocity nitrogen gas source to atomize liquid Sprayed RBCs An external gas atomizer nozzle uses a high velocity nitrogen gas source to atomize liquid media Spraying increases the surface to volume ratio Droplets approximately 10 μm in Sprayed RBCs media. Spraying increases the surface to volume ratio. Droplets, approximately 10 μm in diameter experience mass transfer from the liquid phase into the gas phase resulting in dried diameter, experience mass transfer from the liquid phase into the gas phase, resulting in dried bl dDid d ti t tdf th h t blood. Dried product is extracted from the exhaust gas. N l Nozzle Spectrophotometer Spectrophotometer Microcentrifuge Microcentrifuge Modeling Ai Fl Rt (LPM) St d d Modeling Air Flow Rate (LPM) Standard Spray Dry Testing Spray Dry Testing Terminal Velocity via Stokes’Law Schmidt Number Terminal Velocity via Stokes Law Schmidt Number Figure 1: Percent drying efficiency was derived from mass of Figure 2: The change in absorbance of RBC solution collected sprayed saline both immediately and twelve hours after after spraying as compared to the same solution prior to 2 gd sprayed saline both immediately and twelve hours after i E b t 95% fid after spraying as compared to the same solution prior to i E b t 95% fid p w air gd v air Sc spraying. Error bars represent 95% confidence. spraying. Error bars represent 95% confidence. 18 v Sc D 18 air air AB D Reynolds Number Fröessling Equation A l i dC l i Reynolds Number Fröessling Equation Analysis and Conclusions Heat Exchanger Analysis and Conclusions k d d v Heat Exchanger Spray Nozzle 1 1 3 2 2 0 522 g p k d Re Sc p o air d v Re Funnel Spray Nozzle Nitrogen Source More than 50% of red blood cells lysed in phosphate buffered saline during 2 0.522 Re Sc D Re Syringe and Tube More than 50% of red blood cells lysed in phosphate buffered saline during AB D air S i Syringe and Tube i i Powdered microscope slide drying An additional 10% lysed during rehydration The current Syringe Drying Container RBCs microscope slide drying. An additional 10% lysed during rehydration. The current Diffusion Coefficient Relationship Change in Diameter with Respect to time Pump RBCs t d t 80% f t f lt l ti L i f d bl d ll Diffusion Coefficient Relationship Change in Diameter with Respect to time Pump system removed up to 80% of water from salt solutions. Lysing of red blood cells Fluid Source d d ih d li id fl f 10 L/ i d10 L/ i 3 2 T 2 dd D Fluid Source are reduced with a gas and liquid flow rates of 10 L/min and10 mL/min, T D D 1 1 3 2 2 2 0 522 p AB dd D Re Sc y are reduced with a gas and liquid flow rates of 10 L/min and10 mL/min, 1 AB AB T D D T 2 0.522 w Re Sc y dt d respectively The viscosity and density of the red blood cell solutions sprayed are 1 T w p dt d respectively. The viscosity and density of the red blood cell solutions sprayed are similar to those of water which suggests water can be used to model spraying Red Blood Cell Drying Tests Results The change in particle diameter with similar to those of water, which suggests water can be used to model spraying Red Blood Cell Drying Tests Results The change in particle diameter with properties Red Blood Cell Drying Tests Results respect to time was calculated Using an properties. Red blood cells in phosphate b ffered saline ere placed on standard microscope respect to time was calculated. Using an Red blood cells in phosphate buffered saline were placed on standard microscope iterative method stepwise changes were slides to allo e aporation to occ r Half of the cells l sed d ring dr ing d e to the iterative method, stepwise changes were slides to allow evaporation to occur. Half of the cells lysed during drying due to the made to examine the diameter of the increase in salt concentration After reh dration 40% of original cells remained made to examine the diameter of the L ki F d increase in salt concentration. After rehydration, 40% of original cells remained particle in any instance in time These Looking Forward intact particle in any instance in time. These Looking Forward intact. results are presented in Figure 3 which results are presented in Figure 3, which Further testing should be conducted with human rather than equine red shows that the particles reach steady Further testing should be conducted with human rather than equine red shows that the particles reach steady bl d ll b th ll i i i ifi tl D i d t state diameter of 0 8 μm within 0 3 ms blood cells because the cell size varies significantly. Drying does not state diameter of 0.8 μm within 0.3 ms h d li id dA dd i h ld b after traveling about 6 nm This occurr at the gas and liquid rates tested A second drying stream should be after traveling about 6 nm. This occurr at the gas and liquid rates tested. A second drying stream should be Gas Source suggests an opportunity to increase the investigated Gas Source suggests an opportunity to increase the investigated. ratio of air flow rate to liquid flow rate ratio of air flow rate to liquid flow rate. The model also allows for predictions The model also allows for predictions Figure 3: A graph of the theoretical change in diameter and of how humidity impacts the system distance traveled with respect to time for a liquid flow rate of of how humidity impacts the system, Thanks to Andy Brickman Manfred Dittrich Allyson K Fry Dr Phil H Harding 2 mL/min and an air flow rate of 20 L/min at 27 °C for a 10 which can be further examined Thanks to Andy Brickman, Manfred Dittrich, Allyson K. Fry, Dr. Phil H. Harding, Red blood cells in saline Cells lysing while drying Medium crystallization Rehydration with water 2 mL/min and an air flow rate of 20 L/min at 27 C for a 10 μm water particle which can be further examined. Dr Jill E Parker Marsha C Lampi and Bend Research Red blood cells in saline Cells lysing while drying Medium crystallization Rehydration with water μm water particle Dr. Jill E. Parker, Marsha C. Lampi, and Bend Research.
Transcript of SDifLSpray Drying for Long py yg g -TSt fRdBldCllTerm ... · objective is to explore spray drying...
S D i f L T St f R d Bl d C llSpray Drying for Long Term Storage of Red Blood CellsSpray Drying for Long-Term Storage of Red Blood CellsSpray Drying for Long-Term Storage of Red Blood CellsSpray Drying for Long Term Storage of Red Blood Cellsp y y g g gMeehe Kim Mary McLean John Simeles Dr Adam Higgins (Sponsor)Meehe Kim, Mary McLean, John Simeles, Dr. Adam Higgins (Sponsor)Meehe Kim, Mary McLean, John Simeles, Dr. Adam Higgins (Sponsor), y , , gg ( p )
School of Chemical Biological and Environmental EngineeringSchool of Chemical Biological and Environmental Engineering http://en.wikipedia.org/wiki/File:Erythrocyte_deoxy.jpg1School of Chemical, Biological, and Environmental Engineering, g , g g
M th d S D T t R ltObj ti Methods Spray Dry Test ResultsObjective Methods Spray Dry Test ResultsObjective p y yR d bl d ll d t d f t f i i ft 30 d i th f i t
T t t k t diff t i t i th t d i t ti A 80 °CRed blood cells donated for transfusion expire after 30 days in the refrigerator.
R d Bl d C ll S i Temperature was taken at different points in the system during testing. An 80 °C p y g
Storing and transporting refrigerated blood is costly and inconvenient The Red Blood Cell Suspension p p y g gl t t t t d i t d f t t b it k t
Storing and transporting refrigerated blood is costly and inconvenient. The p nozzle output temperature was used instead of room temperature because it kept objective is to explore spray drying red blood cells as a means of storing cells at p p p pll i t t hil till i i th t f D i ffi i t t d b
objective is to explore spray drying red blood cells as a means of storing cells at cells intact, while still increasing the mass transfer. Drying efficiency was tested by room temperature to improve the logistics of storage , g y g y y
i fl t d li t ti (Fi 1) R d bl d llroom temperature to improve the logistics of storage.
Slide Test varying gas flow rates and saline concentrations (Figure 1). Red blood cells were Slide Test y g g ( g )d th h th l ith h idifi d it t diff ti t th d th dsprayed through the nozzle with humidified nitrogen to differentiate the death due p y g g
t i f th d th d t d i H l bi i l d h d bl dB k d to spraying from the death due to drying. Hemoglobin is released when red blood Background p y g y g gll l d b tifi d b t h t t (Fi 2) Th b bBackground cells lyse and can be quantified by spectrophotometry (Figure 2). The absorbance
Red blood cells are contained by fragile phospholipid membranes and can lyse or fold in Ob i bl dDilute in phosphate
C t ify q y p p y ( g )
t f t d d d t d h l bi i di t d th t 430 i thRed blood cells are contained by fragile phospholipid membranes and can lyse or fold in di i h b l
Obtain bloodp p
buffered salineCentrifuge spectrum of oxygenated and deoxygenated hemoglobin indicated that 430 nm is the conditions shown below. buffered saline
Extractp yg yg g
b t l th t t k t f b th C ll d th t it fl t fExtract best wavelength to take measurements for both. Cell death at nitrogen flow rates of g g10 L/ i i ifi tl l th t 20 L/ i dl f bl d fl tSpray Test 10 L/min are significantly lower than at 20 L/min regardless of blood flow rate.
E lt E tSpray Test g y g
Excess salt Excess waterTemperature < 20 °C Temperature > 80 °Cp p
Red Blood Cell Viability TestingSlow drying time Fast drying time Red Blood Cell Viability TestingSlow drying time Fast drying time
http://www.chrisrepetsky.com/?m=200910
An external gas atomizer nozzle uses a high-velocity nitrogen gas source to atomize liquid Sprayed RBCsAn external gas atomizer nozzle uses a high velocity nitrogen gas source to atomize liquid media Spraying increases the surface to volume ratio Droplets approximately 10 μm in
Sprayed RBCsmedia. Spraying increases the surface to volume ratio. Droplets, approximately 10 μm in diameter experience mass transfer from the liquid phase into the gas phase resulting in drieddiameter, experience mass transfer from the liquid phase into the gas phase, resulting in dried bl d D i d d t i t t d f th h tblood. Dried product is extracted from the exhaust gas. p g
N lNozzleSpectrophotometerSpectrophotometer
MicrocentrifugeMicrocentrifuge
Modeling Ai Fl R t (LPM)St d dModeling Air Flow Rate (LPM)Standard
Spray Dry Testingg ( )
Spray Dry TestingTerminal Velocity via Stokes’ Law Schmidt NumberTerminal Velocity via Stokes Law Schmidt Number Figure 1: Percent drying efficiency was derived from mass of Figure 2: The change in absorbance of RBC solution collectedg y g ysprayed saline both immediately and twelve hours after
g gafter spraying as compared to the same solution prior to
2g d sprayed saline both immediately and twelve hours after
i E b t 95% fidafter spraying as compared to the same solution prior to
i E b t 95% fid p w airg dv
airSc
spraying. Error bars represent 95% confidence. spraying. Error bars represent 95% confidence.
18v
ScD 18 air air ABD
Reynolds Number Fröessling EquationA l i d C l i
Reynolds Number Fröessling EquationAnalysis and ConclusionsHeat Exchanger Analysis and Conclusionsk dd v
Heat ExchangerSpray Nozzle y
11322 0 522g pk d
Re Sc p o aird vRe
Funnel
Spray NozzleNitrogen Source
More than 50% of red blood cells lysed in phosphate buffered saline during2 0.522 Re Sc
D Re
Syringeu e
and Tubeoge Sou ce
More than 50% of red blood cells lysed in phosphate buffered saline during ABDairS i
Syringe and Tubei i
Powdered microscope slide drying An additional 10% lysed during rehydration The currentSyringe Drying Container RBCs microscope slide drying. An additional 10% lysed during rehydration. The current Diffusion Coefficient Relationship Change in Diameter with Respect to time Pump
RBCs p y g y g yt d t 80% f t f lt l ti L i f d bl d ll
Diffusion Coefficient Relationship Change in Diameter with Respect to time Pumpsystem removed up to 80% of water from salt solutions. Lysing of red blood cells Fluid Source y p y g
d d i h d li id fl f 10 L/ i d10 L/ i3
2T 2dd DFluid Source
are reduced with a gas and liquid flow rates of 10 L/min and10 mL/min,TD D
1132
2 2 0 522p ABdd D
Re Sc y are reduced with a gas and liquid flow rates of 10 L/min and10 mL/min, 1AB AB TD D
T
2 0.522 wRe Sc y
dt d
respectively The viscosity and density of the red blood cell solutions sprayed are1T w pdt drespectively. The viscosity and density of the red blood cell solutions sprayed are similar to those of water which suggests water can be used to model sprayingRed Blood Cell Drying Tests ResultsThe change in particle diameter with similar to those of water, which suggests water can be used to model spraying Red Blood Cell Drying Tests ResultsThe change in particle diameter with properties
Red Blood Cell Drying Tests Resultsrespect to time was calculated Using an properties.
Red blood cells in phosphate b ffered saline ere placed on standard microscoperespect to time was calculated. Using an p p
Red blood cells in phosphate buffered saline were placed on standard microscope iterative method stepwise changes were p p p pslides to allo e aporation to occ r Half of the cells l sed d ring dr ing d e to the
iterative method, stepwise changes were slides to allow evaporation to occur. Half of the cells lysed during drying due to the made to examine the diameter of the p y g y gincrease in salt concentration After reh dration 40% of original cells remained
made to examine the diameter of the
L ki F dincrease in salt concentration. After rehydration, 40% of original cells remained particle in any instance in time These Looking Forward
y , gintact
particle in any instance in time. These Looking Forwardintact.results are presented in Figure 3 which gresults are presented in Figure 3, which Further testing should be conducted with human rather than equine redshows that the particles reach steady Further testing should be conducted with human rather than equine red shows that the particles reach steady g qbl d ll b th ll i i i ifi tl D i d tstate diameter of 0 8 µm within 0 3 ms blood cells because the cell size varies significantly. Drying does not state diameter of 0.8 µm within 0.3 ms g y y g
h d li id d A d d i h ld bafter traveling about 6 nm This occurr at the gas and liquid rates tested A second drying stream should beafter traveling about 6 nm. This occurr at the gas and liquid rates tested. A second drying stream should be Gas Sourcesuggests an opportunity to increase the investigatedGas Sourcesuggests an opportunity to increase the investigated.ratio of air flow rate to liquid flow rateratio of air flow rate to liquid flow rate.
The model also allows for predictionsThe model also allows for predictions Figure 3: A graph of the theoretical change in diameter andof how humidity impacts the system g g p gdistance traveled with respect to time for a liquid flow rate ofof how humidity impacts the system,
Thanks to Andy Brickman Manfred Dittrich Allyson K Fry Dr Phil H Hardingp q2 mL/min and an air flow rate of 20 L/min at 27 °C for a 10which can be further examined Thanks to Andy Brickman, Manfred Dittrich, Allyson K. Fry, Dr. Phil H. Harding,
Red blood cells in saline Cells lysing while drying Medium crystallization Rehydration with water2 mL/min and an air flow rate of 20 L/min at 27 C for a 10μm water particle
which can be further examined.Dr Jill E Parker Marsha C Lampi and Bend ResearchRed blood cells in saline Cells lysing while drying Medium crystallization Rehydration with waterμm water particle Dr. Jill E. Parker, Marsha C. Lampi, and Bend Research.
