Using DoE in R&D Projects - statcon.de

Using DoE in R&D ProjectsA Practical Case Study

Ron Stites

Stites & Associates, LLC

Ron Stites

• Former Director of Research for Range Fuels

• Three US Patents in Alternative Fuels

• Analytical Chemist -- BS Chemistry KU

• MBA Finance and Accounting

• Independent Consultant in Research Management – Consulting Since 1996

Email: [email protected]: Ron StitesBlog: http://tek-dev.typepad.com/technology-development/Main Lab: Brighton, CO, USA

Context

• Industrial R&D– Questions are Fairly Well Defined

– Results Can be Translated into Financial Benefit

– An “Application” NOT “Pure Science”

– Cost Well Monitored

– Cost versus Financial Benefit a Frequent Discussion

– Generally in the Area of “Sustaining Technology” and not “Disruptive Technology”

• Evolutionary or

• Revolutionary

For Society – Success of Industrial R&D as Critical as Academic and Government Sponsored R&D

The Unique Nature of R&D Projects

• In General – Many Unknowns

– Key Factors

– Key Results

– Causal Relationships

– Value of Solution

– Real Budget

– Real Time Line

– Real Goals

Technical

Programmatic

It is often the Case that there is LESS Time and Money than

Officially Budgeted

Where to Start?

• Problem/Opportunity Definition– What is the Purpose of the Project?

– What does a Successful Project Look Like?

– What Factors are Important?

– What Results Need to be Measured?

– What is the Necessary Time-Line?

– What has been Done in the Past?• Internal

• External

– Can We Put into Writing Something that Seems “Do-able?”• Research Plan

• Project Charter/Plan

This is an Intuitive Process that is Messy but MUST NOT be Skipped

What Next?

• Evaluate the Factors and Results Desired– Do Measurement Techniques Exist?– Do We Know the Ranges of Interest?– Do We Know the Required Accuracy and Precision?

• Two Separate Questions

• Can We State Measureable Hypotheses About the Relationships Between Factors and Results?

• Are We Ready for a DoE?– What are the Gaps?

In Many Cases the Answer is NO!Often New (or Modified) Measurement Techniques are Needed

These Must be Developed and Tested – Again in a more Intuitive Fashion

Guidance for Starting from Scratch

• Keep it Simple• Keep it Cheap

– Avoid Going Directly to the Lab – Go to the Library First– Avoid Buying the “Quantum Electromechanical Nuclear Pile” to

Measure pH – Use pH Paper to Start– Don’t Spend More than about 25% of Budget on First Try

• Keep it Moving Forward– Gets Bits of Knowledge at a Time– Keep Trying Things and Taking Good Notes

• Keep Evaluating– What are the Gaps?– Be Honest – Even Cynical about what You “KNOW”

Case Study

The Situation:• Industrial Equipment Supplier• Selling an Existing Devise into a “New” Market

– Corn Ethanol– Not Much Known About Corn Ethanol Technology

• Devise is Known to Mix and Heat Slurries Well• Anecdotal Evidence that the Devise “Improves Ethanol

Yield”– Some Inconsistency in Data – “Art”– Expensive to Test at Plant

• Hope to Learn How the Devise Works to Improve Consistency of Performance

Preliminary Work

Learn About Corn Ethanol Technology

• Starch to Glucose to Ethanol– NOT Sugar Directly to Ethanol

• Key Opportunity is in Breakdown of Starch

• Key Gap is Measuring the Breakdown of Starch to Glucose– Stepwise

• Starch to Maltodextrins

• Maltodextrins to Glucose

OH

OH

O

H

H

H

H

H

HO

CH2OH

~O

6

54

3 2 1

Many 1-4 α Glucoside Bonds

OH

OH

O

H

H

H

H

H

HO

CH2OH

HO

6

54

3 2 1

OH

O

O

H

H

H

H

H

HO

CH2OH

~O

6

54

3 2 1

OH

OH

O

H

H

H

H

H

HO

CH2OH6

54

3 2 1

- H2O (Dehydration)

Forming Long Chains Results in Starch

Amylose in Particular (DP 300 to 3000)

Only 1 “Reducing End” per Chain

OH

OH

O

H

H

H

H

H

HO

CH2OH

~O

6

54

3 2 1

Additional 1-6 α Bond Branching

OH

O

O

H

H

H

H

H

HO

CH2OH

~O

6

54

3 2 1

OH

O~

O

H

H

H

H

H

HO

CH2OH6

54

3 2 1

- H2O (Dehydration)

This Branching Results in Amylopectin another form

of Starch(DP 2,000 to 200,000)

Yeast Cannot Digest These Macromolecules

They Must be Broken Down at Least to Maltose but Much

Preferred is Glucose

Basic Process

Grind

React with Amylase and

GlucoamylaseEnzymes

(~85C)

Cook(~70C)

Ferment(~32C)

Distill

• Process More Complex Than Sugarcane• Initial Cook is Called “Slurrying”• Holding at Temperature is “Liquefaction”• Enzymes Present Throughout

What is the Opportunity?

• Speedup the Hydrolysis• Use Less Enzymes• Minimize “Burning” of the Starch

– Shorter Chain Sugars Can React Together and with Proteins to Form Polymers that Yeast Cannot Digest – Some are Even Toxic to Yeast

• Monitored by “DP” Analysis of the Maltodextrins• But Measurement of Maltodextrins is Not Common – “New”

Methods are Needed– Specialized Techniques are NOT Available in the Plant – Samples Must

be Sent to a Lab

• BUT WORSE – Samples are NOT Stable – Hydrolysis Continues

This Careful Thinking Resulted in a Very Specific R&D ProjectHow to Preserve Samples for Maltodextrin Analysis

This Took Several Months to Accomplish

Clear Problem Definition

Is There a Practical Way to Treat Samples to Stop the Hydrolysis Reaction so that Samples Can be Shipped to a Lab for “DP” Analysis?

– What Methods are “Practical?”

– What Methods do not “Change” the “DP Profile?”

– What Lab Methods are Available and Practical?• Internal?

• External?

“Library” Work

• Reviewed Published Literature on DP Analysis of Starch– Considerable Work Done in Corn Ethanol Industry

– Much More Done in Starch Industry

• Contacted Plants

• Contacted Engineering Firms

• Contacted Analytical Suppliers

• Contacted Academic Researchers

• Contacted Enzyme Suppliers

Found Clear Guidance on DP Analysis – HPLC AnalysisFound A Number of Good Hypotheses to Chase – pH & Temp

Did NOT Find a Complete Testing ProtocolTwo Parallel Paths – Develop HPLC & Experimental Process

HPLC

min0 10 20 30 40 50

Norm.

0

100000

200000

300000

400000

500000

600000

RID1 A, Refractive Index Signal (20120222\DP000002.D)

10.

492

15.

226

- D

P12

+

20.

304

- D

P8

21.

601

- D

P7

22.

978

- D

P6

25.

014

- D

P5

27.

516

- D

P4

30.

634

- D

P3

34.

921

- D

P2

41.

920

- D

P1

44.

772

58.

582

min0 10 20 30 40 50

Norm.

0

50000

100000

150000

200000

250000

300000

350000


10.

768

11.

647

15.

027

- D

P12

+

17.

434

- D

P11

18.

232

- D

P10

19.

126

- D

P9

20.

309

- D

P8

21.

707

- D

P7

23.

076

- D

P6

25.

111

- D

P5

27.

580

- D

P4

30.

688

- D

P3

34.

966

- D

P2

41.

948

- D

P1

44.

916

47.

699

- G

lyce

rol

57.

428

Corn Mash “Slurried”“0 Hrs”

Corn Mash Liquified“2.5 Hrs”

Lab Liquefaction Difficult to Repeat

Result Depends on:• Time• Temperature• Mixing• Water Content

Needed a Consistent Starting Material to Measure Changes (Actually Lack of Change) Reliably

Gave Up on Corn – Too Difficult to Reproduce

Finally Worked Out a Surrogate

Purchased Maltodextrin Looked Much Like Corn SlurryContained Large DP’s and Some Small DP’s

Easy to Use and Control – Took a Week to Work Out

min0 10 20 30 40 50

Norm.

0

20000

40000

60000

80000

100000

120000

140000


12.

537

14.

616

- D

P12

+

18.

359

- D

P10

19.

252

- D

P9

20.

439

21.

730

- D

P7

23.

097

- D

P6

25.

138

- D

P5

27.

670

- D

P4

30.

751

- D

P3

35.

057

- D

P2

39.

244

42.

010

- D

P1

5% Maltodextrin Solution

Protocol Development

• Make Up Consistent Maltodextrin Solution (10%)

• Add Controlled Amount of Enzyme

• Monitor %DP<9 as a Measure of Reaction

• Treat Some with 26% Sulfuric Acid (pH’s from 1 to 3)

• Treat Some with Cooling (0 and 25 C)

• Treat Some with Both

• Hold at a Typical Shipping Times (24 to 70 hr)

Several Types of Tests were Run Over 2 WeeksSort of “Played Around” to See What Made Sense

The Pay-Off

• After About 4 Weeks it Became Clear– pH Was the Most Important

• pH of 2 Appeared to Work and Not Create Degradation of Sugars

– Temperature Might be Important• Not as Clear Cut – Not Sure About Time versus Temp• Thought that holding at a higher temp for a few hours might help

– Maltodextrin Appeared to be a Useful Surrogate• Reacted Rapidly with Enzyme• Easy to Dissolve in Water at a Consistent Concentration (by Mass)

– The HPLC Method Easily Detected Changes • Changes in Smaller Sugars Concentrations (%</DP9) Easily

Measured at Times and Temperatures of Interest

We Seemed to Have Closed the Gaps Preventing a Useful DoE

The Actual DoE

After About 1 Week of “Playing Around” a Useful DoE was Actually Fairly Simple

Factor 1 Factor 2 Factor 3 Response 1

Std Run A:pH B:Incubate at 60C

C:Store

Temperature % <DP9

Units pH C %

3 1 2 No 3 29.22%

6 2 5.5 Yes 25 77.17%

8 3 5.5 No 25 77.45%

5 4 2 Yes 25 24.49%

1 5 2 Yes 3 25.20%

7 6 2 No 25 25.04%

4 7 5.5 No 3 64.99%

2 8 5.5 Yes 3 74.31%

Results Pretty Convincing

Most Important Factor is A = pH

Incubation Slightly Worse

We Chose the lnTransformation because it

Worked Well for Other Reaction Rate

Experiments – Slight Improvement Here

Design-Expert® SoftwareLn(% <DP9)

Shapiro-Wilk testW-value = 0.943p-value = 0.686A: pHB: Incubate at 60CC: Store Temperature

Positive Effects Negative Effects

Half-Normal Plot

Ha

lf-N

orm

al %

Pro

ba

bili

ty

|Standardized Effect|

0.00 0.10 0.21 0.31 0.42 0.52 0.62 0.73 0.83 0.94 1.04

0

10

20

30

50

70

80

90

95

A

FIGURE XI – HALF NORMAL PLOT OF lnTRANSFORMED DATA

Additional Evaluation

Adjusting pH Has Profound Effect – As Expected


A: pHB: Incubate at 60CC: Store Temperature

Positive Effects Negative Effects

Pareto Chart

t-V

alu

e o

f |E

ffect

|

Rank

0.00

3.00

6.01

9.01

12.02

15.02

18.03

1 2 3 4 5 6 7

Bonferroni Limit 3.99706

t-Value Limit 2.44691

A

FIGURE XII – PARETO CHART OF EFFECTS

Does It “Preserve”?

Only pH Adjustment and Storage at 3C has Change Less Than the Standard Deviation of the HPLC Method (~1%)

Since These are Practical Things to Do in the Field, This was the Method Selected – Took About 6 Weeks

Sample %<DP9 %Delta Notes

SX 20-0 29.14% Mother Liquor

SX 20-1 29.22% 0.28% pH 2, No Incubation, Store 3C

SX 20-4 24.49% -15.94% pH 2, Incubate, Store 25C

SX 20-5 25.20% -13.51% pH 2, Incubate, Store 3C

SX 20-6 25.04% -14.06% pH 2, No Incubation, Store 25C

Epilog

• Method Implemented• Seemed to Work for Slurries and Liquefacts• Some “Strange” Results with Fermentation Samples – Most

of the Carbohydrates Gone• Discovered that the SO4(-2) was Interfering with the DP12+

Peak Throwing Off Calculations• Method Modified to Remove SO4(-2) by Treatment with

Ba(OH)2 and Filtering• Now Seems to Work Well with Ferms Also

Always Remain Skeptical and Vigilent – Especially When Applying to Other Conditions and Samples

Questions?

Stites Awarded Patents

• US Patent 7,884,253 B2, 2/8/2011; Methods And Apparatus For Selectively Producing Ethanol From Synthesis Gas; Stites and Hohman.

• US Patent 7,919,070 B2, Apr. 5, 2011; Multi-Zone Reforming Methods and Apparatus for Conversion of Devolatilized Biomass to Syngas; Stites, Biehle, Klepper and Ridley.

• US Patent 8,142,530 B2, Mar. 27, 2012; Methods and Apparatus for Producing Syngas and Alcohols; Klepper, Geerstsema, Tirmizi, Robota and Stites.

Additional Patents in Various Stages of Application/Approval

HPLC Trace versus Time

0

1

2

3

4

5

6

7

DP12+ DP11 DP10 DP9 DP8 DP7 DP6 DP5 DP4 DP3 DP2 DP1

Malto-Dextrins (SX19)32C pH 5.5

Time 0

Time 0.5

Time 1.5

Time 2.0

Time 4.0

Time 22

Time 48

Results Pretty Convincing

We Chose the lnTransformation because it

Worked Well for Other Reaction Rate

Experiments – Not Really Needed Here FIGURE XI – HALF NORMAL PLOT OF ln

TRANSFORMED DATA


LambdaCurrent = 0Best = -0.02Low C.I. = -1.32High C.I. = 1.27

Recommend transform:None (Lambda = 1)

Lambda

Ln(R

esi

dual

SS

)

Box-Cox Plot for Power Transforms

-5.00

-4.50

-4.00

-3.50

-3.00

-2.50

-3 -2 -1 0 1 2 3

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Using DoE in R&D Projects - statcon.de