NREL is a national laboratory of the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.

High Throughput Pretreatment and Enzyme 

Hydrolysis of Biomass: Screening Recalcitrance in 

Large Sample Populations 

Presented by

Stephen R. Decker

1st International Conference on Lignocellulosic Ethanol

Copenhagen, Denmark

October 13-15, 2010

NREL/PR-2700-49599
BioEnergy Science Center

Oak Ridge 

National Laboratory

University of Georgia

University of Tennessee

Dartmouth College

Georgia Institute of Technology

West Virginia University

ArborGen, LLC

Ceres, Incorporated 

Mascoma Corporation

Verenium Corporation

Samuel Roberts Noble Foundation

National Renewable

Energy Laboratory

Brookhaven National

Laboratory

University of California–

Riverside 

Cornell University 

Washington State University

University of Minnesota 

North Carolina State University

Virginia Polytechnic Institute

University of California–Los Angeles

A multi-institutional, DOE-funded, Bioenergy Research Center 

dedicated to understanding and modifying Biomass Recalcitrance

322 people 

in 20 institutions
Somerville et al

Biomass Recalcitrance

• Plant cell walls are complex, heterogenous structures

– Three major polymers

• Cellulose, hemicellulose (multiple types), and lignin

• Components closely associated and differentially reactive

– Cell wall polysaccharides are structural

• More recalcitrant than storage carbohydrates such as starch

– Steps to overcome recalcitrance

• Pretreatment- enables efficient enzymatic hydrolysis

• Enzymatic hydrolysis is less damaging than pretreatment

• Recalcitrance- genetic and environmental factors

– Must screen large populations to find and correlate 

underlying principles and effects

– Affects pretreatability and enzyme digestibility

Genetics

Process

Conventional

enzyme fermentation

Ethanol

Butanol 

Alcohols

Fermentation

Recalcitrance

Consolidated

bioprocessing

Chemical

catalysis

Hydrocarbons

Synthetic

biology

Hydrocarbons

Environment
Screening Large Populations for 

Recalcitrance Variants

10s of 

samples

grams of 

material

Weeks of 

analysis

100s/1000s 

of samples

milligrams 

of material

Hours of 

analysis

How is Recalcitrance Affected by:

•Environmental variables

•Natural diversity

•Genetic manipulation

•Enzyme composition

•Pretreatment chemistry

•Pretreatment severity

Repeat many times

Fast- many challenges

Slow- huge effort required
Barriers to High Throughput Screening 

in Biomass Conversion

• Defined by large sample sets

– Manual labor in collecting, milling, extracting, starch 

removal

– Demands small-scale, automated processing

• Biomass is insoluble, heterogeneous

– Accurate dispensing, distribution is difficult

– Static is major problem

• Severe pretreatment conditions

– High temperatures (150-200oC), pressures, low pH 

• Solids/Liquids separation

– Small scale losses large impact on analysis

• Enzyme systems are complex, slow

– Long conversion times (days, not minutes)

• Sugar product detection 

– HPLC is slow (one plate = 2 days of HPLC time) 

• Chemical analysis/composition

– Standard techniques too slow

• Systematically addressed/constrained the 

barriers

100 mi

200 km

Skagit (Sedro Woolley)

Skykomish (Monroe)

Puyallup (Orting) Columbia 

(Longview)

Existing collections (N = 500; 1–2 trees/site)

New collections (N = 580; 140–160 trees/site)
Biomass Preparation and Dispensing

• Collect and mill by hand

• De-starch/extract in “teabags” 

– Wash extensively, extract in Soxhlet

• Solids- Symyx MTM Powdernium (2)

– Dispense/track quantities

• Automated, 0.1 mg precision

– Multiple substrate capacity 

• 144 or 200 deck capacity

• 15 plate capacity

• Liquid- Beckman-Coulter Biomek FX

– Catalyst, enzymes, sugar assays
96-Well Pretreatment Reactor Plate 

7

• Single piece reactor plate

– Sequential pretreatment/enzymatic saccharification

– No Solid-Liquid separation- neutralize only

• Outer dimensions of standard 96-well plate

– Robot friendly

• Larger well diameter increases volume capacity

• 117 steam/cooling water channels per plate

– 4 channels/well 

– even heating and cooling

• Ni/Au plated wells add corrosion resistance

– Hastalloy, other coatings option for dilute acid

• Stackable to enable multi-plate pretreatments

• Custom designed and manufactured

– Available from Aspen Machining, Lakewood, CO, USA

Steam/

cooling water

Parr Reactor

Head Plate

Base Plate

Reactor plates

Teflon gaskets

Al foil seal

Nuts/bevel 

washers

Tightening studs
HTP Plate Clamping System Design Maximum 20 Plates Top Plate Bottom Plate

• Stacked system

– 20 plates, 1920 pretreatments

• Aluminum head and base plates 

– Fit snug in Parr reactor 

– designed for rigidity

• Plate sealing

– 0.020” glass-reinforced PFTE 

gaskets 

– Aluminum foil seals on plates

• Steam/cooling water enters 

through bottom clamping plate

• Even heating well-to-well and 

vertically

0

20

40

60

80

100

120

140

160

180

200

0 1 2 3

Temperature (o

C)

Minutes

Reactor Plate Heating Rate

Well G2

Well D6

Well C11

Closing up Parr Reactor

Steam start

vent closed
y = 0.8382x - 0.0789

R2 = 0.9958

0

0.5

1

1.5

2

2.5

3

0.0 1.0 2.0 3.0 4.0

Xylose by HPLC (mg/mL)

Xylose by XDH (mg/mL)

Comparison of Xylose Quantitation Methods

y = 1.0198x + 0.0584

R2 = 0.9796

0

0.5

1

1.5

2

2.5

3

0.0 0.5 1.0 1.5 2.0 2.5 3.0

Glucose by HPLC (mg/mL)

Glucose by GOPOD (mg/mL)

Comparison of Glucose Detection Methods

Evaporation Control-Sugar Quantitation • 3 days at 40oC

• Clear adhesive seal was inconsistent 

– Edge effects

• Used magnets in steam ports 

– Press fit into aluminum base plate

– Thin magnetic stainless steel top plate

Glucose - glucose oxidase/peroxidase

D-Glucose + O2 + H2O  D-gluconate + H2O2

H2O2 + p-HBA + 4-aminoantipyrine  quinoneimine dye (A510)

• GOPOD cross-reactivity

– Xylose, arabinose, 

xylobiose- none

– Galactose, mannosetrace

Xylose - ß-D-xylose dehydrogenase

ß-D-Xylose + NAD+  D-xylonic acid + NADH (A340) + H+

• XDH cross-reactivity

– Xylose, arabinose, 

mannose- none

– Galactose, trace

– Xylobiose- moderate

0

20

40

60

80

100

120

% Water Remaining in Plate

0 50 100 150

Time (h)

Water Loss Throughout Incubation

Teflon-coated plates, 40oC, static

1. Plate + seal only

2. Plate + Seal + 13 magnets and steel plates
Parallel Plate System Overview

10

Stack Plates

Pretreat

Load Plates

Add Catalyst and 

Seal

Disassemble, add 

enzymes, buffer

Seal,

Incubate

Enzyme Mediated 

Sugar Detection

Robotics Systems

Enable High Volume

Custom Plates 

and Reactor
Analysis of Cell Wall Chemistry

• MBMS- rapid estimate of cell wall chemistry

– High information content

– Complements wet chemistry

• PCA of MS from ~1500 greenhouse poplar. 

– Black = low nitrogen

– Purple = high nitrogen. 

• The two clusters indicate differing cell wall 

chemistry 

– Different lignin composition

11

-8

-6

-4

-2

0

2

4

6

-8 -6 -4 -2 0 2 4 6 8 10 12

 RESULT1, X-expl: 62%,13% 

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UF325-2-CHEM UF325-2-CHEM

UF325-5-CHEM UF325-5-CHEM

UF325-6-CHEM UF325-6-CHEM

UF326-1-CHEM

UF326-1-CHEM

UF326-2-CHEM UF326-1-CHEM

UF326-2-CHEM

UF326-5-CHEM

UF326-5-CHEM

UF326-6-CHEM

UF327-1-CHEM

UF327-1-CHEM

UF327-2-CHEM

UF327-2-CHEM

UF327-5-CHEM

UF327-5-CHEM

UF327-6-CHEM

UF327-6-CHEM

UF328-1-CHEM

UF328-1-CHEM

UF328-2-CHEM

UF328-2-CHEM

UF328-5-CHEM

UF328-5-CHEM UF328-6-CHEM UF328-6-CHEM UF337-1-CHEM

UF337-1-CHEM

UF337-2-CHEM UF337-2-CHEM

UF337-5-CHEM

UF337-6-CHEM

UF337-6-CHEM

UF343-1-CHEM

UF343-1-CHEM

UF343-2-CHEM

UF343-2-CHEM

UF343-5-CHEM

UF343-5-CHEM

UF343-6-CHEM

UF343-6-CHEM

UF344-1-CHEM

UF344-1-CHEM

UF344-2-CHEM

UF344-2-CHEM

UF344-5-CHEM

UF344-5-CHEM

UF344-6-CHEM

UF344-6-CHEM

UF345-1-CHEM

UF345-1-CHEM

UF345-2-CHEM

UF345-2-CHEM

UF345-5-CHEM

UF345-5-CHEM

UF35-1-CHEM

UF35-1-CHEM

UF35-2-CHEM

UF35-5-CHEM UF35-5-CHEM

UF35-6-CHEM UF35-6-CHEM

UF352-1-CHEM

UF352-1-CHEM

UF352-2-CHEM

UF352-2-CHEM

UF352-5-CHEM

UF352-5-CHEM

UF352-6-CHEM

UF352-6-CHEM

UF359-1-CHEM

UF359-2-CHEM UF359-1-CHEM

UF359-2-CHEM

UF359-5-CHEM

UF359-5-CHEM

UF359-6-CHEM

UF359-6-CHEM

UF36-1-CHEM

UF36-1-CHEM

UF36-2-CHEM UF36-2-CHEM

UF36-5-CHEM

UF36-5-CHEM

UF36-6-CHEM

UF36-6-CHEM

UF361-1-CHEM

UF361-2-CHEM UF361-1-CHEM

UF361-2-CHEM

UF361-5-CHEM

UF361-5-CHEM

UF361-6-CHEM

UF361-6-CHEM

UF366-1-CHEM

UF366-1-CHEM

UF366-1-CHEM

UF366-2-CHEM

UF366-2-CHEM

UF366-5-CHEM

UF366-5-CHEM UF366-6-CHEM

UF366-6-CHEM

UF368-1-CHEM

UF368-1-CHEM

UF368-1-CHEM

UF368-2-CHEM

UF368-2-CHEM

UF368-5-CHEM

UF368-5-CHEM

UF368-6-CHEM

UF368-6-CHEM

UF370-1-CHEM

UF370-1-CHEM

UF370-2-CHEM

UF370-2-CHEM

UF370-5-CHEM

UF370-5-CHEM

UF370-6-CHEM

UF370-6-CHEM

UF372-1-CHEM

UF372-1-CHEM

UF372-2-CHEM

UF372-2-CHEM

UF372-5-CHEM

UF372-5-CHEM

UF372-6-CHEM

UF372-6-CHEM UF38-2-CHEM UF38-2-CHEM

UF38-5-CHEM

UF38-5-CHEM

UF38-6-CHEM

UF38-6-CHEM

UF380-1-CHEM

UF380-1-CHEM

UF380-2-CHEM UF380-2-CHEM

UF380-5-CHEM

UF380-5-CHEM

UF380-6-CHEM

UF380-6-CHEM

UF381-1-CHEM

UF381-1-CHEM

UF381-2-CHEM UF381-2-CHEM

UF381-5-CHEM

UF381-5-CHEM

UF381-6-CHEM

UF381-6-CHEM

UF382-1-CHEM

UF382-1-CHEM

UF382-2-CHEM

UF382-2-CHEM

UF382-5-CHEM

UF382-5-CHEM

UF382-6-CHEM UF382-6-CHEM

UF383-1-CHEM

UF383-1-CHEM

UF383-2-CHEM

UF383-2-CHEM

UF383-5-CHEM

UF383-6-CHEM UF383-5-CHEM

UF383-6-CHEM

UF385-1-CHEM

UF385-1-CHEM

UF385-2-CHEM UF385-2-CHEM

UF385-5-CHEM

UF385-5-CHEM

UF385-6-CHEM

UF385-6-CHEM

UF389-1-CHEM

UF389-1-CHEM

UF389-1-CHEM

UF389-2-CHEM UF389-2-CHEM

UF389-5-CHEM

UF389-5-CHEM

UF389-6-CHEM

UF389-6-CHEM

UF39-1-CHEM

UF39-1-CHEM

UF39-2-CHEM

UF39-5-CHEM

UF39-6-CHEM UF39-5-CHEM UF39-6-CHEM

UF390-1-CHEM

UF390-1-CHEM

UF390-2-CHEM

UF390-2-CHEM

UF392-1-CHEM UF392-1-CHEM

UF392-2-CHEM UF392-2-CHEM

UF392-5-CHEM

UF392-5-CHEM UF392-6-CHEM

UF394-1-CHEM

UF394-1-CHEM

UF394-2-CHEM

UF394-2-CHEM

UF394-5-CHEM

UF394-5-CHEM

UF394-6-CHEM

UF394-6-CHEM

UF395-1-CHEM

UF395-1-CHEM

UF395-5-CHEM

UF395-5-CHEM

UF395-6-CHEM

UF396-1-CHEM

UF396-1-CHEM

UF396-2-CHEM

UF396-2-CHEM

UF396-5-CHEM

UF396-5-CHEM

UF396-6-CHEM UF396-6-CHEM

UF397-1-CHEM

UF397-1-CHEM

UF397-1-CHEM

UF397-2-CHEM

UF397-2-CHEM

UF397-5-CHEM

UF397-5-CHEM

UF397-6-CHEM

UF397-6-CHEM

UF398-1-CHEM

UF398-1-CHEM

UF398-2-CHEM

UF398-2-CHEM

UF398-5-CHEM

UF398-6-CHEM UF398-5-CHEM

UF398-6-CHEM

UF40-1-CHEM

UF40-1-CHEM

UF40-2-CHEM UF40-2-CHEM

UF40-5-CHEM

UF40-5-CHEM

UF40-6-CHEM

UF40-6-CHEM

UF401-1-CHEM

UF401-1-CHEM

UF401-2-CHEM

UF401-2-CHEM

UF401-5-CHEM

UF401-5-CHEM

UF402-1-CHEM UF402-1-CHEM

UF402-2-CHEM UF402-2-CHEM

UF402-5-CHEM

UF402-5-CHEM

UF402-6-CHEM

UF402-6-CHEM

UF404-1-CHEM

UF404-1-CHEM UF404-2-CHEM

UF404-2-CHEM

UF404-5-CHEM

UF404-5-CHEM

UF404-6-CHEM UF404-6-CHEM

UF405-1-CHEM

UF405-1-CHEM

UF405-1-CHEM

UF405-2-CHEM UF405-2-CHEM

UF405-5-CHEM

UF405-5-CHEM

UF405-6-CHEM UF405-6-CHEM

UF406-1-CHEM

UF406-1-CHEM

UF406-2-CHEM

UF406-2-CHEM

UF406-5-CHEM

UF406-5-CHEM UF406-6-CHEM UF406-6-CHEM

UF407-1-CHEM

UF407-1-CHEM UF407-2-CHEM

UF407-2-CHEM

UF407-5-CHEM

UF407-5-CHEM

UF407-6-CHEM

UF407-6-CHEM

UF408-1-CHEM

UF408-1-CHEM

UF408-2-CHEM

UF408-2-CHEM

UF408-5-CHEM

UF408-5-CHEM

UF408-6-CHEM

UF408-6-CHEM

UF409-1-CHEM

UF409-1-CHEM

UF409-2-CHEM

UF409-2-CHEM

UF409-5-CHEM

UF409-5-CHEM

UF409-6-CHEM

UF409-6-CHEM

UF417-1-CHEM

UF417-1-CHEM

UF417-2-CHEM

UF417-2-CHEM

UF417-5-CHEM

UF417-5-CHEM

UF417-6-CHEM

UF417-6-CHEM

UF418-1-CHEM

UF418-1-CHEM UF418-1-CHEM

UF418-2-CHEM

UF418-2-CHEM

UF418-5-CHEM

UF418-5-CHEM

UF418-6-CHEM

UF418-6-CHEM

UF419-1-CHEM

UF419-1-CHEM

UF419-2-CHEM UF419-2-CHEM

UF419-5-CHEM

UF419-5-CHEM

UF419-6-CHEM

UF419-6-CHEM

UF42-1-CHEM

UF42-1-CHEM

UF42-2-CHEM UF42-2-CHEM

UF42-5-CHEM

UF42-5-CHEM UF42-6-CHEM

UF42-6-CHEM

UF421-5-CHEM

UF421-6-CHEM UF421-5-CHEM UF421-6-CHEM

UF435-1-CHEM

UF435-1-CHEM UF435-2-CHEM UF435-1-CHEM

UF435-5-CHEM

UF435-6-CHEM UF435-5-CHEM

UF435-6-CHEM

UF436-1-CHEM

UF436-1-CHEM

UF436-2-CHEM

UF436-2-CHEM

UF436-5-CHEM

UF436-5-CHEM

UF436-6-CHEM UF436-6-CHEM

UF437-2-CHEM

UF437-2-CHEM

UF437-5-CHEM

UF437-5-CHEM

UF437-6-CHEM

UF440-1-CHEM

UF440-1-CHEM

UF440-2-CHEM UF440-2-CHEM

UF440-5-CHEM

UF440-6-CHEM UF440-5-CHEM

UF441-1-CHEM

UF441-1-CHEM

UF441-2-CHEM

UF441-2-CHEM

UF441-5-CHEM

UF441-5-CHEM

UF441-6-CHEM

UF441-6-CHEM UF445-1-CHEM

UF445-1-CHEM

UF445-1-CHEM

UF445-2-CHEM

UF445-2-CHEM

UF445-5-CHEM

UF445-5-CHEM

UF445-6-CHEM

UF445-6-CHEM

UF451-1-CHEM

UF451-2-CHEM UF451-1-CHEM UF451-2-CHEM

UF451-5-CHEM

UF451-5-CHEM

UF451-6-CHEM

UF452-1-CHEM

UF452-1-CHEM

UF452-2-CHEM

UF452-2-CHEM

UF452-5-CHEM

UF452-5-CHEM

UF452-6-CHEM UF452-6-CHEM

UF453-1-CHEM

UF453-1-CHEM UF453-2-CHEM UF453-2-CHEM

UF453-5-CHEM

UF453-5-CHEM

UF453-6-CHEM

UF453-6-CHEM

UF456-1-CHEM

UF456-1-CHEM UF456-2-CHEM

UF456-2-CHEM

UF456-5-CHEM

UF456-5-CHEM

UF456-6-CHEM

UF457-1-CHEM UF457-1-CHEM

UF457-2-CHEM

UF457-2-CHEM

UF457-5-CHEM

UF457-5-CHEM

UF457-6-CHEM

UF46-1-CHEM

UF46-1-CHEM

UF46-2-CHEM

UF46-5-CHEM

UF46-5-CHEM

UF46-6-CHEM

UF46-6-CHEM

UF460-2-CHEM

UF460-2-CHEM

UF460-5-CHEM

UF460-5-CHEM

UF460-6-CHEM

UF460-6-CHEM

UF461-1-CHEM

UF461-1-CHEM

UF461-2-CHEM

UF461-2-CHEM

UF461-5-CHEM

UF461-5-CHEM UF461-6-CHEM

UF461-6-CHEM

UF464-1-CHEM

UF464-1-CHEM

UF464-2-CHEM

UF464-2-CHEM

UF464-5-CHEM

UF464-5-CHEM

UF464-6-CHEM

UF464-6-CHEM

UF465-1-CHEM

UF465-1-CHEM

UF465-2-CHEM UF465-2-CHEM

UF465-5-CHEM

UF465-5-CHEM

UF465-6-CHEM UF465-6-CHEM

UF467-1-CHEM

UF467-1-CHEM

UF467-2-CHEM

UF467-2-CHEM

UF467-5-CHEM

UF467-5-CHEM

UF467-6-CHEM

UF467-6-CHEM

UF468-1-CHEM

UF468-1-CHEM

UF468-2-CHEM

UF468-2-CHEM

UF468-5-CHEM

UF468-5-CHEM

UF468-6-CHEM

UF468-6-CHEM

UF47-1-CHEM

UF47-1-CHEM

UF47-1-CHEM

UF47-2-CHEM

UF47-2-CHEM

UF473-2-CHEM

UF473-2-CHEM

UF473-5-CHEM

UF473-5-CHEM

UF473-6-CHEM UF473-6-CHEM

UF476-5-CHEM UF476-5-CHEM

UF476-6-CHEM UF476-6-CHEM

UF477-1-CHEM

UF477-1-CHEM

UF477-2-CHEM

UF477-2-CHEM

UF477-5-CHEM

UF477-5-CHEM

UF477-6-CHEM

UF477-6-CHEM

UF479-2-CHEM UF479-2-CHEM

UF479-5-CHEM

UF479-5-CHEM

UF479-6-CHEM

UF479-6-CHEM

UF481-1-CHEM

UF481-1-CHEM

UF481-2-CHEM UF481-2-CHEM

UF481-5-CHEM

UF481-5-CHEM UF481-6-CHEM

UF481-6-CHEM

UF488-1-CHEM

UF488-2-CHEM UF488-1-CHEM

UF488-5-CHEM

UF488-5-CHEM

UF488-6-CHEM

UF488-6-CHEM

UF489-1-CHEM

UF489-1-CHEM UF489-2-CHEM

UF489-2-CHEM

UF489-6-CHEM UF489-6-CHEM

UF49-1-CHEM

UF49-1-CHEM

UF49-2-CHEM UF49-2-CHEM

UF49-5-CHEM

UF49-5-CHEM

UF49-6-CHEM UF49-6-CHEM

UF490-1-CHEM

UF490-1-CHEM

UF490-2-CHEM

UF490-2-CHEM

UF490-5-CHEM

UF490-5-CHEM

UF490-6-CHEM

UF490-6-CHEM

UF491-1-CHEM

UF491-1-CHEM

UF491-2-CHEM UF491-2-CHEM

UF491-5-CHEM

UF491-5-CHEM

UF491-6-CHEM

UF491-6-CHEM

UF492-1-CHEM

UF492-1-CHEM

UF492-2-CHEM

UF492-2-CHEM

UF492-5-CHEM

UF492-5-CHEM

UF492-6-CHEM

UF492-6-CHEM

UF493-1-CHEM

UF493-1-CHEM

UF493-2-CHEM

UF493-2-CHEM

UF493-5-CHEM

UF493-5-CHEM

UF493-6-CHEM

UF493-6-CHEM

UF494-1-CHEM

UF494-1-CHEM

UF494-1-CHEM

UF494-2-CHEM UF494-2-CHEM

UF494-5-CHEM

UF494-5-CHEM

UF494-6-CHEM

UF494-6-CHEM

UF497-1-CHEM

UF497-1-CHEM

UF497-2-CHEM UF497-2-CHEM

UF497-5-CHEM

UF497-5-CHEM

UF497-6-CHEM

UF497-6-CHEM

UF498-1-CHEM

UF498-1-CHEM

UF498-1-CHEM

UF498-2-CHEM

UF498-2-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-6-CHEM UF498-6-CHEM

UF499-1-CHEM

UF499-1-CHEM

UF499-2-CHEM UF499-2-CHEM

UF499-5-CHEM

UF499-5-CHEM UF499-6-CHEM

UF499-6-CHEM

UF500-1-CHEM

UF500-1-CHEM

UF500-2-CHEM UF500-2-CHEM

UF500-5-CHEM

UF500-5-CHEM

UF500-6-CHEM

UF500-6-CHEM

UF503-1-CHEM

UF503-1-CHEM

UF503-2-CHEM

UF503-2-CHEM

UF503-6-CHEM UF503-6-CHEM

UF505-1-CHEM

UF505-1-CHEM

UF505-2-CHEM UF505-2-CHEM

UF505-5-CHEM

UF505-5-CHEM

UF505-6-CHEM

UF505-6-CHEM

UF512-1-CHEM

UF512-1-CHEM

UF512-5-CHEM

UF512-6-CHEM UF512-5-CHEM UF512-6-CHEM

UF513-1-CHEM

UF513-1-CHEM

UF513-2-CHEM

UF513-2-CHEM

UF513-5-CHEM

UF513-5-CHEM

UF513-6-CHEM UF513-6-CHEM

UF515-1-CHEM

UF515-1-CHEM

UF515-5-CHEM

UF515-5-CHEM

UF515-6-CHEM UF515-6-CHEM

UF516-1-CHEM

UF516-1-CHEM

UF516-2-CHEM

UF516-2-CHEM

UF516-5-CHEM

UF516-5-CHEM

UF516-6-CHEM UF516-6-CHEM

UF518-1-CHEM

UF518-1-CHEM

UF518-2-CHEM

UF518-2-CHEM

UF518-5-CHEM

UF518-6-CHEM UF518-5-CHEM

UF518-6-CHEM

UF525-1-CHEM

UF525-1-CHEM

UF525-1-CHEM

UF525-2-CHEM

UF525-2-CHEM

UF526-1-CHEM

UF526-1-CHEM

UF526-2-CHEM

UF526-2-CHEM

UF526-5-CHEM

UF526-5-CHEM

UF526-6-CHEM

UF526-6-CHEM

UF529-1-CHEM

UF529-1-CHEM

UF529-2-CHEM

UF529-2-CHEM

UF529-5-CHEM

UF529-5-CHEM

UF529-6-CHEM UF529-6-CHEM

UF53-1-CHEM

UF53-1-CHEM

UF53-2-CHEM UF53-2-CHEM

UF53-5-CHEM

UF53-5-CHEM

UF53-6-CHEM UF53-6-CHEM

UF532-1-CHEM

UF532-1-CHEM UF532-2-CHEM

UF532-5-CHEM

UF532-6-CHEMUF532-5-CHEM

UF532-6-CHEM

UF533-1-CHEM UF533-1-CHEM

UF533-2-CHEM

UF533-2-CHEM

UF533-5-CHEM

UF533-5-CHEM

UF533-6-CHEM UF533-6-CHEM

UF534-1-CHEM

UF534-1-CHEM

UF534-2-CHEM

UF534-2-CHEM

UF534-5-CHEM

UF534-5-CHEM UF534-6-CHEM

UF535-1-CHEM

UF535-1-CHEM

UF535-2-CHEM

UF535-2-CHEM

UF535-5-CHEM

UF535-5-CHEM

UF535-6-CHEM UF535-6-CHEM

UF537-1-CHEM

UF537-1-CHEM

UF537-2-CHEM UF537-2-CHEM

UF537-5-CHEM

UF537-5-CHEM

UF537-6-CHEM

UF537-6-CHEM

UF538-1-CHEM

UF538-1-CHEM UF538-2-CHEM

UF538-2-CHEM

UF538-5-CHEM

UF538-5-CHEM

UF538-6-CHEM

UF539-1-CHEM

UF539-1-CHEM

UF539-2-CHEM

UF539-2-CHEM

UF539-5-CHEM

UF539-5-CHEM

UF539-6-CHEM

UF54-1-CHEM

UF54-1-CHEM

UF54-2-CHEM

UF54-5-CHEM

UF54-5-CHEM UF54-6-CHEM UF54-6-CHEM

UF541-1-CHEM

UF541-1-CHEM

UF541-2-CHEM

UF541-2-CHEM

UF541-5-CHEM

UF541-5-CHEM

UF541-6-CHEM UF541-6-CHEM

UF542-1-CHEM UF542-1-CHEM

UF542-2-CHEM

UF542-5-CHEM

UF542-5-CHEM

UF542-6-CHEM

UF542-6-CHEM

UF543-2-CHEM

UF543-2-CHEM

UF544-1-CHEM

UF544-1-CHEM

UF544-2-CHEM UF544-2-CHEM

UF544-5-CHEM

UF544-5-CHEM

UF544-6-CHEM

UF544-6-CHEM

UF546-1-CHEM

UF546-1-CHEM

UF546-2-CHEM UF546-2-CHEM

UF546-5-CHEM

UF546-5-CHEM

UF546-6-CHEM

UF546-6-CHEM

UF547-1-CHEM

UF547-1-CHEM

UF547-2-CHEM

UF547-5-CHEM

UF547-5-CHEM

UF547-6-CHEM

UF552-1-CHEM UF552-1-CHEM

UF552-2-CHEM

UF552-2-CHEM

UF552-5-CHEM UF552-5-CHEM

UF552-6-CHEM

UF552-6-CHEM

UF554-1-CHEM

UF554-1-CHEM

UF554-2-CHEM

UF554-2-CHEM

UF554-5-CHEM

UF554-5-CHEM

UF554-6-CHEM

UF554-6-CHEM

UF56-1-CHEM

UF56-1-CHEM

UF56-2-CHEM

UF56-2-CHEM

UF56-5-CHEM

UF56-5-CHEM

UF56-6-CHEM UF56-6-CHEM

UF561-1-CHEM

UF561-1-CHEM

UF561-2-CHEM UF561-2-CHEM

UF561-5-CHEM UF561-5-CHEM

UF561-6-CHEM UF561-6-CHEM UF562-1-CHEM

UF562-1-CHEM

UF562-2-CHEM UF562-2-CHEM

UF562-5-CHEM

UF562-5-CHEM

UF562-6-CHEM

UF562-6-CHEM

UF565-1-CHEM UF565-1-CHEM

UF565-2-CHEM

UF565-2-CHEM

UF565-5-CHEM

UF565-5-CHEM

UF565-6-CHEM

UF565-6-CHEM

UF567-1-CHEM

UF567-1-CHEM

UF567-2-CHEM UF567-2-CHEM

UF567-5-CHEM

UF567-5-CHEM

UF567-6-CHEM UF567-6-CHEM

UF569-2-CHEM UF569-2-CHEM

UF569-5-CHEM

UF569-5-CHEM

UF569-6-CHEM UF570-1-CHEM

UF570-1-CHEM

UF570-2-CHEM

UF570-2-CHEM

UF570-5-CHEM

UF570-5-CHEM

UF570-6-CHEM

UF570-6-CHEM

UF571-1-CHEM

UF571-1-CHEM

UF571-1-CHEM

UF571-2-CHEM

UF571-2-CHEM

UF572-2-CHEM

UF572-2-CHEM

UF572-5-CHEM UF572-5-CHEM

UF572-6-CHEM UF572-6-CHEM

UF576-1-CHEM

UF576-1-CHEM

UF576-2-CHEM

UF576-2-CHEM

UF576-5-CHEM

UF576-5-CHEM

UF576-6-CHEM

UF576-6-CHEM

UF578-5-CHEM

UF578-5-CHEM

UF578-6-CHEM UF578-6-CHEM UF579-1-CHEM

UF579-1-CHEM

UF579-2-CHEM

UF579-2-CHEM

UF579-5-CHEM

UF579-5-CHEM

UF58-1-CHEM UF58-1-CHEM

UF58-2-CHEM

UF58-2-CHEM

UF58-5-CHEM

UF58-5-CHEM

UF58-6-CHEM

UF58-6-CHEM

UF581-1-CHEM

UF581-1-CHEM

UF581-2-CHEM

UF581-2-CHEM

UF581-5-CHEM

UF581-5-CHEM

UF581-6-CHEM

UF581-6-CHEM

UF582-1-CHEM

UF582-1-CHEM

UF582-2-CHEM

UF582-2-CHEM

UF582-5-CHEM

UF582-5-CHEM

UF582-6-CHEM

UF582-6-CHEM

UF584-1-CHEM

UF584-1-CHEM

UF584-2-CHEM

UF584-2-CHEM

UF584-5-CHEM

UF584-5-CHEM UF584-6-CHEM

UF585-1-CHEM

UF585-1-CHEM

UF585-2-CHEM

UF585-2-CHEM

UF585-5-CHEM

UF585-5-CHEM UF585-6-CHEM UF585-6-CHEM

UF586-1-CHEM

UF586-1-CHEM

UF586-2-CHEM

UF586-2-CHEM

UF586-5-CHEM

UF586-5-CHEM

UF586-6-CHEM

UF586-6-CHEM

UF587-1-CHEM

UF587-1-CHEM

UF587-2-CHEM

UF587-2-CHEM

UF587-5-CHEM

UF587-5-CHEM

UF587-6-CHEM

UF587-6-CHEM

UF588-1-CHEM

UF588-1-CHEM

UF588-2-CHEM UF588-2-CHEM

UF588-5-CHEM

UF588-5-CHEM

UF588-6-CHEM UF588-6-CHEM

UF589-1-CHEM

UF589-1-CHEM

UF589-2-CHEM

UF589-2-CHEM

UF589-5-CHEM

UF589-5-CHEM

UF589-6-CHEM

UF59-1-CHEM

UF59-1-CHEM UF59-2-CHEM

UF59-2-CHEM

UF59-5-CHEM

UF59-5-CHEM

UF59-6-CHEM

UF59-6-CHEM

UF590-1-CHEM

UF590-1-CHEM

UF590-2-CHEM

UF590-2-CHEM

UF590-5-CHEM

UF590-5-CHEM

UF590-6-CHEM UF590-6-CHEM

UF596-2-CHEM

UF596-2-CHEM

UF596-5-CHEM

UF596-5-CHEM

UF596-6-CHEM

UF596-6-CHEM

UF600-1-CHEM

UF600-1-CHEM

UF600-2-CHEM UF600-1-CHEM

UF600-2-CHEM

UF600-5-CHEM

UF600-5-CHEM

UF600-6-CHEM UF600-6-CHEM

UF61-1-CHEM

UF61-1-CHEM

UF61-2-CHEM

UF61-2-CHEM

UF61-5-CHEM

UF61-5-CHEM

UF61-6-CHEM

UF61-6-CHEM

UF62-1-CHEM

UF62-1-CHEM

UF62-2-CHEM UF62-2-CHEM

UF62-5-CHEM

UF62-5-CHEM

UF62-6-CHEM UF62-6-CHEM

UF65-1-CHEM

UF65-1-CHEM

UF65-2-CHEM

UF65-2-CHEM

UF66-1-CHEM

UF66-1-CHEM

UF66-2-CHEM UF66-2-CHEM

UF66-5-CHEM

UF66-5-CHEM

UF66-6-CHEM

UF66-6-CHEM

UF68-1-CHEM UF68-1-CHEM

UF68-2-CHEM

UF68-2-CHEM

UF68-5-CHEM

UF68-5-CHEM

UF68-6-CHEM

UF68-6-CHEM

UF69-1-CHEM

UF69-1-CHEM

UF69-2-CHEM

UF69-2-CHEM

UF69-5-CHEM

UF69-5-CHEM

UF69-6-CHEM UF69-6-CHEM

UF7-1-CHEM

UF7-1-CHEM UF7-2-CHEM

UF7-2-CHEM

UF7-5-CHEM

UF7-5-CHEM

UF7-6-CHEM

UF7-6-CHEM

UF70-1-CHEM UF70-1-CHEM

UF70-2-CHEMUF70-2-CHEM

UF70-5-CHEM

UF70-5-CHEM

UF70-6-CHEM

UF70-6-CHEM

UF700-1-CHEM

UF700-1-CHEM

UF700-2-CHEM

UF700-2-CHEM

UF700-5-CHEM

UF700-5-CHEM UF700-6-CHEM

UF700-6-CHEM

UF701-1-CHEM UF701-1-CHEM

UF701-2-CHEM UF701-2-CHEM

UF701-5-CHEM

UF701-5-CHEM

UF701-6-CHEM

UF703-1-CHEM

UF703-1-CHEM

UF703-2-CHEM

UF703-2-CHEM

UF703-5-CHEM

UF703-5-CHEM

UF703-6-CHEM UF703-6-CHEM

UF705-1-CHEM

UF705-1-CHEM UF705-1-CHEM UF705-2-CHEM

UF705-2-CHEM

UF705-5-CHEM

UF705-5-CHEM

UF705-6-CHEM

UF705-6-CHEM

UF706-1-CHEM

UF706-1-CHEM

UF706-2-CHEM

UF706-2-CHEM

UF706-5-CHEM

UF706-5-CHEM

UF706-6-CHEM UF706-6-CHEM

UF707-1-CHEM

UF707-1-CHEM

UF707-2-CHEM UF707-2-CHEM

UF707-5-CHEM

UF707-5-CHEM

UF707-6-CHEM

UF707-6-CHEM

UF709-1-CHEM

UF709-1-CHEM

UF709-2-CHEM

UF709-2-CHEM

UF709-5-CHEM

UF709-5-CHEM

UF709-6-CHEM

UF709-6-CHEM

UF710-1-CHEM

UF710-1-CHEM

UF710-1-CHEM

UF710-2-CHEM UF710-2-CHEM

UF710-5-CHEM

UF710-5-CHEM

UF710-6-CHEM

UF710-6-CHEM

UF714-1-CHEM

UF714-1-CHEM

UF714-1-CHEM

UF714-2-CHEM

UF714-2-CHEM

UF714-5-CHEM

UF714-5-CHEM UF714-6-CHEM

UF714-6-CHEM

UF715-1-CHEM

UF715-1-CHEM

UF715-1-CHEM

UF715-2-CHEM UF715-2-CHEM

UF715-5-CHEM

UF715-5-CHEM

UF715-6-CHEM

UF715-6-CHEM

UF716-1-CHEM

UF716-1-CHEM

UF716-2-CHEM UF716-2-CHEM

UF716-5-CHEM

UF716-5-CHEM

UF716-6-CHEM UF716-6-CHEM

UF717-1-CHEM

UF717-1-CHEM

UF717-2-CHEM UF717-2-CHEM

UF717-5-CHEM

UF717-5-CHEM

UF717-6-CHEM

UF717-6-CHEM

UF718-1-CHEM

UF718-1-CHEM

UF718-2-CHEM

UF718-2-CHEM

UF718-5-CHEM

UF718-5-CHEM

UF718-6-CHEM

UF718-6-CHEM

UF720-1-CHEM

UF720-1-CHEM

UF720-2-CHEM

UF720-2-CHEM

UF720-5-CHEM

UF720-5-CHEM

UF720-6-CHEM

UF721-1-CHEM

UF721-1-CHEM

UF721-2-CHEM

UF721-2-CHEM

UF722-1-CHEM

UF722-1-CHEM

UF722-2-CHEM

UF722-2-CHEM

UF722-5-CHEM

UF722-5-CHEM

UF722-6-CHEM

UF722-6-CHEM

UF723-1-CHEM

UF723-1-CHEM

UF723-2-CHEM

UF723-2-CHEM

UF723-5-CHEM

UF723-5-CHEM

UF723-6-CHEM UF723-6-CHEM

UF724-1-CHEM

UF724-2-CHEM UF724-1-CHEM

UF724-2-CHEM

UF724-5-CHEM

UF724-5-CHEM

UF724-6-CHEM

UF727-1-CHEM

UF727-1-CHEM

UF727-2-CHEM

UF727-5-CHEM

UF727-5-CHEM

UF727-6-CHEM

UF727-6-CHEM

UF728-5-CHEM

UF728-5-CHEM

UF728-6-CHEM

UF729-1-CHEM

UF729-1-CHEM

UF729-2-CHEM

UF729-2-CHEM

UF729-5-CHEM

UF729-5-CHEM

UF729-6-CHEM

UF729-6-CHEM

UF73-1-CHEM

UF73-1-CHEM

UF73-2-CHEM

UF73-2-CHEM UF73-5-CHEM

UF73-6-CHEM UF73-5-CHEM

UF73-6-CHEM

UF731-1-CHEM

UF731-1-CHEM

UF731-2-CHEM

UF731-2-CHEM

UF731-5-CHEM

UF731-6-CHEM UF731-5-CHEM

UF731-6-CHEM

UF734-1-CHEM

UF734-2-CHEM UF734-1-CHEM

UF734-2-CHEM

UF734-5-CHEM

UF734-5-CHEM

UF734-6-CHEM

UF734-6-CHEM

UF735-1-CHEM

UF735-1-CHEM

UF735-2-CHEM UF735-2-CHEM

UF735-5-CHEM

UF735-5-CHEM

UF735-6-CHEM

UF735-6-CHEM

UF74-1-CHEM

UF74-1-CHEM

UF74-2-CHEM

UF74-2-CHEM

UF74-5-CHEM

UF74-5-CHEM

UF74-6-CHEM

UF74-6-CHEM UF741-1-CHEM

UF741-1-CHEM

UF741-2-CHEM

UF741-2-CHEM

UF743-1-CHEM

UF743-1-CHEM

UF743-2-CHEM

UF743-2-CHEM

UF743-5-CHEM

UF743-5-CHEM

UF743-6-CHEM

UF743-6-CHEM

UF745-1-CHEM

UF745-2-CHEM UF745-2-CHEM UF745-1-CHEM

UF745-5-CHEM

UF745-5-CHEM

UF745-6-CHEM UF745-6-CHEM

UF748-1-CHEM

UF748-1-CHEM

UF748-2-CHEM

UF748-2-CHEM

UF748-5-CHEM

UF748-5-CHEM

UF748-6-CHEM

UF748-6-CHEM

UF751-1-CHEM

UF751-1-CHEM

UF751-2-CHEM UF751-2-CHEM UF751-5-CHEM UF751-5-CHEM

UF751-6-CHEM UF751-6-CHEM

UF754-1-CHEM

UF754-1-CHEM

UF754-2-CHEM

UF754-2-CHEM

UF754-5-CHEM

UF754-5-CHEM

UF754-6-CHEM

UF754-6-CHEM

UF755-1-CHEM

UF755-1-CHEM

UF755-2-CHEM UF755-2-CHEM

UF755-5-CHEM UF755-5-CHEM

UF755-6-CHEM

UF78-1-CHEM

UF78-1-CHEM

UF78-2-CHEM

UF78-2-CHEM

UF78-5-CHEM

UF78-5-CHEM UF78-6-CHEM

UF78-6-CHEM

UF79-1-CHEM

UF79-1-CHEM

UF79-2-CHEM

UF79-2-CHEM

UF79-5-CHEM

UF79-6-CHEM UF79-5-CHEM

UF79-6-CHEM

UF800-1-CHEM UF800-1-CHEM

UF800-2-CHEM

UF800-2-CHEM

UF800-5-CHEM

UF800-5-CHEM

UF800-6-CHEM

UF800-6-CHEM

UF801-1-CHEM

UF801-2-CHEM UF801-1-CHEM

UF801-2-CHEM

UF801-5-CHEM

UF801-5-CHEM

UF801-6-CHEM UF801-6-CHEM

UF802-1-CHEM

UF802-1-CHEM

UF802-2-CHEM UF802-2-CHEM

UF802-5-CHEM

UF802-5-CHEM

UF802-6-CHEM

UF802-6-CHEM

UF803-1-CHEM

UF803-1-CHEM

UF803-2-CHEM

UF803-2-CHEM

UF803-5-CHEM

UF803-5-CHEM

UF803-6-CHEM

UF803-6-CHEM

UF804-1-CHEM

UF804-1-CHEM

UF804-2-CHEM

UF804-2-CHEM

UF804-5-CHEM

UF804-5-CHEM

UF804-6-CHEM

UF804-6-CHEM

UF805-1-CHEM

UF805-1-CHEM

UF805-2-CHEM

UF805-2-CHEM

UF805-5-CHEM

UF805-5-CHEM

UF805-6-CHEM

UF805-6-CHEM

UF806-1-CHEM

UF806-2-CHEM UF806-1-CHEM

UF806-5-CHEM

UF806-5-CHEM

UF806-6-CHEM

UF806-6-CHEM

UF807-1-CHEM

UF807-1-CHEM

UF807-2-CHEM UF807-2-CHEM

UF807-5-CHEM

UF807-5-CHEM

UF807-6-CHEM UF807-6-CHEM

UF808-1-CHEM

UF808-1-CHEM

UF808-1-CHEM

UF808-2-CHEM UF808-2-CHEM UF808-5-CHEM

UF808-5-CHEM

UF808-6-CHEM

UF808-6-CHEM

UF809-1-CHEM

UF809-1-CHEM

UF809-2-CHEM

UF809-5-CHEM

UF809-5-CHEM

UF809-6-CHEM

UF809-6-CHEM

UF810-1-CHEM

UF810-1-CHEM

UF810-5-CHEM

UF810-5-CHEM

UF810-6-CHEM UF810-6-CHEM

UF811-1-CHEM

UF811-1-CHEM

UF811-2-CHEM UF811-2-CHEM UF812-1-CHEM

UF812-1-CHEM

UF812-2-CHEM

UF812-2-CHEM

UF812-5-CHEM

UF812-5-CHEM

UF812-6-CHEM

UF813-1-CHEM

UF813-1-CHEM UF813-2-CHEM

UF813-2-CHEM

UF813-5-CHEM

UF813-5-CHEM

UF813-6-CHEM UF813-6-CHEM

UF814-1-CHEM

UF814-1-CHEM UF814-2-CHEM

UF814-2-CHEM

UF814-5-CHEM

UF814-5-CHEM

UF814-6-CHEM

UF814-6-CHEM

UF815-1-CHEM

UF815-1-CHEM

UF815-2-CHEM

UF815-2-CHEM

UF815-5-CHEM

UF815-5-CHEM

UF815-6-CHEM UF815-6-CHEM

UF816-1-CHEM

UF816-1-CHEM

UF816-2-CHEM

UF816-2-CHEM

UF816-5-CHEM

UF816-5-CHEM

UF816-6-CHEM

UF816-6-CHEM

UF817-5-CHEM

UF817-5-CHEM

UF817-6-CHEM

UF817-6-CHEM

UF818-1-CHEM

UF818-1-CHEM

UF818-2-CHEM

UF818-2-CHEM

UF818-5-CHEM

UF818-5-CHEM

UF818-6-CHEM

UF818-6-CHEM

UF819-1-CHEM

UF819-1-CHEM

UF819-2-CHEM UF819-2-CHEM

UF819-5-CHEM

UF819-5-CHEM

UF819-6-CHEM

UF819-6-CHEM

UF820-1-CHEM

UF820-1-CHEM

UF820-2-CHEM

UF820-2-CHEM

UF820-5-CHEM

UF820-5-CHEM

UF820-6-CHEM UF820-6-CHEM

UF821-2-CHEM

UF821-2-CHEM

UF821-5-CHEM

UF821-5-CHEM

UF821-6-CHEM UF821-6-CHEM

UF822-1-CHEM

UF822-1-CHEM

UF822-2-CHEM UF822-2-CHEM

UF822-5-CHEM

UF822-5-CHEM

UF822-6-CHEM

UF822-6-CHEM

UF823-1-CHEM

UF823-1-CHEM UF823-1-CHEM UF823-2-CHEM UF823-2-CHEM

UF823-5-CHEM

UF823-5-CHEM

UF823-6-CHEM

UF823-6-CHEM

UF824-1-CHEM

UF824-1-CHEM

UF824-2-CHEM

UF824-2-CHEM

UF824-5-CHEM

UF824-5-CHEM

UF824-6-CHEM UF824-6-CHEM

UF825-1-CHEM

UF825-1-CHEM

UF825-2-CHEM UF825-2-CHEM

UF825-5-CHEM

UF825-5-CHEM UF825-6-CHEM

UF825-6-CHEM

UF826-1-CHEM

UF826-1-CHEM

UF826-2-CHEM UF826-2-CHEM

UF826-6-CHEM

UF826-6-CHEM

UF827-1-CHEM

UF827-1-CHEM

UF827-2-CHEM

UF827-2-CHEM

UF827-5-CHEM

UF827-5-CHEM

UF827-6-CHEM UF827-6-CHEM

UF828-1-CHEM

UF828-1-CHEM

UF828-2-CHEM

UF828-2-CHEM

UF829-1-CHEM

UF829-1-CHEM

UF829-2-CHEM

UF829-2-CHEM

UF829-5-CHEM

UF829-5-CHEM

UF829-6-CHEM

UF829-6-CHEM

UF830-5-CHEM

UF830-5-CHEM

UF830-6-CHEM UF830-6-CHEM

UF831-1-CHEM

UF831-1-CHEM

UF831-2-CHEM

UF831-2-CHEM

UF831-5-CHEM

UF831-5-CHEM

UF831-6-CHEM

UF831-6-CHEM

UF832-1-CHEM

UF832-1-CHEM

UF832-2-CHEM

UF832-2-CHEM

UF832-5-CHEM

UF832-5-CHEM

UF832-6-CHEM UF832-6-CHEM

UF833-1-CHEM

UF833-1-CHEM

UF833-2-CHEM UF833-2-CHEM

UF833-5-CHEM

UF833-5-CHEM

UF833-6-CHEM UF833-6-CHEM

UF834-2-CHEM UF834-2-CHEM

UF834-5-CHEM

UF834-5-CHEM

UF834-6-CHEM

UF834-6-CHEM

UF835-1-CHEM

UF835-1-CHEM

UF835-2-CHEM UF835-2-CHEM

UF835-5-CHEM

UF835-5-CHEM

UF835-6-CHEM UF835-6-CHEM

UF836-1-CHEM

UF836-1-CHEM

UF836-1-CHEM

UF836-2-CHEM

UF836-2-CHEM

UF836-5-CHEM

UF836-5-CHEM

UF836-6-CHEM

UF836-6-CHEM

UF837-1-CHEM UF837-1-CHEM

UF837-2-CHEM

UF837-2-CHEM

UF837-5-CHEM

UF837-5-CHEM

UF837-6-CHEM UF837-6-CHEM

UF838-1-CHEM

UF838-1-CHEM

UF838-2-CHEM UF838-2-CHEM

UF838-5-CHEM

UF838-5-CHEM

UF838-6-CHEM UF838-6-CHEM

UF839-1-CHEM

UF839-1-CHEM

UF839-2-CHEM

UF839-2-CHEM

UF839-5-CHEM

UF839-5-CHEM

UF839-6-CHEM

UF839-6-CHEM UF840-1-CHEM

UF840-1-CHEM

UF840-2-CHEM

UF840-2-CHEM UF840-5-CHEM

UF840-5-CHEM

UF840-6-CHEM UF840-6-CHEM

UF841-1-CHEM

UF841-1-CHEM

UF841-2-CHEM UF841-2-CHEM

UF841-5-CHEM

UF841-5-CHEM

UF841-6-CHEM

UF841-6-CHEM

UF842-1-CHEM

UF842-2-CHEM UF842-1-CHEM UF842-1-CHEM

UF842-5-CHEM

UF842-5-CHEM

UF842-6-CHEM

UF842-6-CHEM

UF844-1-CHEM

UF844-1-CHEM UF844-2-CHEM UF844-2-CHEM

UF844-5-CHEM

UF844-5-CHEM

UF844-6-CHEM

UF844-6-CHEM

UF845-1-CHEM

UF845-1-CHEM

UF845-2-CHEM

UF845-2-CHEM

UF845-5-CHEM

UF845-5-CHEM

UF845-6-CHEM

UF845-6-CHEM

UF846-1-CHEM UF846-1-CHEM

UF846-1-CHEM

UF846-2-CHEM

UF846-2-CHEM

UF846-5-CHEM

UF846-5-CHEM

UF846-6-CHEM

UF846-6-CHEM

UF847-1-CHEM

UF847-1-CHEM

UF847-2-CHEM

UF847-2-CHEM

UF847-5-CHEM

UF847-5-CHEM

UF848-1-CHEM

UF848-1-CHEM

UF848-2-CHEM UF848-2-CHEM

UF848-5-CHEM

UF848-5-CHEM

UF848-6-CHEM

UF848-6-CHEM

UF849-1-CHEM

UF849-1-CHEM

UF849-2-CHEM UF849-2-CHEM

UF849-5-CHEM

UF849-5-CHEM

UF849-6-CHEM UF849-6-CHEM

UF850-1-CHEM

UF850-1-CHEM

UF850-2-CHEM

UF850-2-CHEM

UF850-5-CHEM

UF850-5-CHEM

UF850-6-CHEM

UF850-6-CHEM

UF852-1-CHEM

UF852-1-CHEM

UF852-2-CHEM

UF852-2-CHEM

UF852-5-CHEM

UF852-5-CHEM

UF852-6-CHEM

UF852-6-CHEM

UF853-1-CHEM

UF853-1-CHEM

UF853-2-CHEM

UF853-2-CHEM

UF853-5-CHEM

UF853-5-CHEM

UF853-6-CHEM UF853-6-CHEM

UF854-1-CHEM

UF854-1-CHEM

UF854-2-CHEM

UF854-2-CHEM

UF854-5-CHEM

UF854-5-CHEM

UF854-6-CHEM

UF854-6-CHEM

UF855-1-CHEM

UF855-1-CHEM

UF855-2-CHEM

UF855-2-CHEM

UF855-5-CHEM

UF855-5-CHEM

UF855-6-CHEM UF855-6-CHEM

UF856-1-CHEM

UF856-1-CHEM

UF856-2-CHEM

UF856-2-CHEM

UF856-5-CHEM

UF856-5-CHEM

UF856-6-CHEM

UF856-6-CHEM UF857-1-CHEM

UF857-1-CHEM

UF857-2-CHEM

UF857-2-CHEM

UF857-5-CHEM

UF857-5-CHEM

UF857-6-CHEM

UF857-6-CHEM

UF858-1-CHEM

UF858-2-CHEM UF858-1-CHEM

UF858-2-CHEM

UF858-5-CHEM

UF858-6-CHEM UF858-5-CHEM

UF859-1-CHEM

UF859-1-CHEM

UF859-2-CHEM UF859-2-CHEM

UF859-5-CHEM

UF859-5-CHEM

UF859-6-CHEM

UF860-1-CHEM

UF860-1-CHEM

UF860-2-CHEM UF860-2-CHEM

UF860-5-CHEM

UF860-5-CHEM

UF860-6-CHEM

UF860-6-CHEM

UF861-1-CHEM

UF861-1-CHEM

UF861-2-CHEM

UF861-2-CHEM

UF861-5-CHEM

UF861-5-CHEM

UF861-6-CHEM UF861-6-CHEM

UF862-1-CHEM

UF862-1-CHEM

UF862-2-CHEM

UF862-2-CHEM

UF862-5-CHEM

UF862-5-CHEM

UF862-6-CHEM

UF862-6-CHEM

UF863-1-CHEM

UF863-1-CHEM

UF863-2-CHEM UF863-2-CHEM

UF863-5-CHEM

UF863-5-CHEM UF863-6-CHEM

UF863-6-CHEM

UF864-1-CHEM

UF864-1-CHEM

UF864-2-CHEM

UF864-2-CHEM

UF864-5-CHEM

UF864-5-CHEM UF864-6-CHEM

UF864-6-CHEM

UF865-1-CHEM

UF865-1-CHEM

UF865-2-CHEM

UF865-2-CHEM

UF865-5-CHEM

UF865-5-CHEM

UF865-6-CHEM UF865-6-CHEM

UF866-1-CHEM

UF866-1-CHEM

UF866-2-CHEM

UF866-2-CHEM

UF867-1-CHEM

UF867-1-CHEM

UF867-1-CHEM

UF867-2-CHEM

UF867-2-CHEM

UF867-5-CHEM

UF867-5-CHEM UF867-6-CHEM UF867-6-CHEM

UF868-1-CHEM

UF868-1-CHEM

UF868-2-CHEM

UF868-2-CHEM

UF868-5-CHEM

UF868-5-CHEM UF868-6-CHEM

UF868-6-CHEM

UF869-1-CHEM

UF869-1-CHEM

UF869-2-CHEM

UF869-2-CHEM

UF869-5-CHEM

UF869-5-CHEM

UF869-6-CHEM

UF869-6-CHEM UF870-1-CHEM

UF870-2-CHEM UF870-1-CHEM UF870-2-CHEM UF870-5-CHEM

UF870-6-CHEM UF870-6-CHEM UF870-5-CHEM

UF871-1-CHEM

UF871-1-CHEM

UF871-2-CHEM

UF871-2-CHEM

UF871-5-CHEM

UF871-5-CHEM

UF871-6-CHEM

UF871-6-CHEM

UF872-1-CHEM

UF872-1-CHEM

UF872-2-CHEM

UF872-2-CHEM

UF872-5-CHEM

UF872-5-CHEM

UF872-6-CHEM UF872-6-CHEM

UF873-1-CHEM

UF873-1-CHEM

UF873-1-CHEM UF873-2-CHEM UF873-2-CHEM

UF873-5-CHEM

UF873-5-CHEM

UF873-6-CHEM

UF873-6-CHEM

UF874-1-CHEM

UF874-1-CHEM

UF874-2-CHEM UF874-2-CHEM

UF874-5-CHEM

UF874-5-CHEM

UF874-6-CHEM

UF874-6-CHEM

UF875-1-CHEM

UF875-1-CHEM

UF875-2-CHEM

UF875-2-CHEM

UF875-5-CHEM

UF875-5-CHEM

UF875-6-CHEM

UF875-6-CHEM

UF876-1-CHEM

UF876-1-CHEM

UF876-1-CHEM

UF876-2-CHEM UF876-2-CHEM

UF876-5-CHEM

UF876-5-CHEM

UF876-6-CHEM

UF876-6-CHEM

UF877-1-CHEM

UF877-1-CHEM

UF877-1-CHEM

UF877-2-CHEM

UF877-2-CHEM

UF877-5-CHEM

UF877-5-CHEM

UF877-6-CHEM UF877-6-CHEM

UF878-1-CHEM

UF878-1-CHEM

UF878-2-CHEM

UF878-2-CHEM

UF878-5-CHEM

UF878-5-CHEM

UF878-6-CHEM UF878-6-CHEM

UF879-1-CHEM

UF879-1-CHEM

UF879-2-CHEM

UF879-2-CHEM

UF879-5-CHEM

UF879-5-CHEM

UF879-6-CHEM

UF879-6-CHEM

UF880-1-CHEM

UF880-1-CHEM

UF880-2-CHEM

UF880-2-CHEM

UF880-5-CHEM

UF880-5-CHEM

UF880-6-CHEM

UF880-6-CHEM

UF881-1-CHEM

UF881-1-CHEM

UF881-2-CHEM UF881-2-CHEM

UF881-5-CHEM

UF881-5-CHEM

UF881-6-CHEM

UF881-6-CHEM

UF882-1-CHEM

UF882-2-CHEM UF882-1-CHEM UF882-2-CHEM

UF882-5-CHEM

UF882-5-CHEM

UF882-6-CHEM

UF882-6-CHEM

UF883-1-CHEM

UF883-1-CHEM

UF883-2-CHEM

UF883-5-CHEM

UF883-5-CHEM

UF883-6-CHEM

UF883-6-CHEM

UF884-1-CHEM

UF884-1-CHEM

UF884-2-CHEM

UF884-2-CHEM

UF884-5-CHEM

UF884-5-CHEM

UF884-6-CHEM UF884-6-CHEM

UF885-1-CHEM

UF885-1-CHEM

UF885-2-CHEM

UF885-2-CHEM

UF885-5-CHEM

UF885-5-CHEM

UF885-6-CHEM

UF885-6-CHEM

UF886-1-CHEM

UF886-1-CHEM

UF886-2-CHEM

UF886-2-CHEM

UF886-5-CHEM

UF886-5-CHEM

UF886-6-CHEM UF886-6-CHEM

UF887-1-CHEM

UF887-1-CHEM

UF887-2-CHEM UF887-2-CHEM

UF887-5-CHEM

UF887-5-CHEM

UF887-6-CHEM

UF887-6-CHEM

UF888-1-CHEM UF888-1-CHEM

UF888-2-CHEM

UF888-2-CHEM

UF888-5-CHEM

UF888-5-CHEM

UF888-6-CHEM

UF888-6-CHEM

UF889-1-CHEM

UF889-1-CHEM

UF889-2-CHEM

UF889-2-CHEM

UF889-5-CHEM

UF889-5-CHEM

UF889-6-CHEM UF889-6-CHEM

UF890-1-CHEM

UF890-1-CHEM

UF890-2-CHEM

UF890-5-CHEM

UF890-5-CHEM

UF890-6-CHEM

UF891-1-CHEM

UF891-1-CHEM

UF891-1-CHEM

UF891-2-CHEM

UF891-2-CHEM

UF891-5-CHEM

UF891-5-CHEM

UF891-6-CHEM

UF891-6-CHEM

UF892-1-CHEM

UF892-1-CHEM

UF892-2-CHEM UF892-2-CHEM

UF892-5-CHEM

UF892-5-CHEM

UF892-6-CHEM UF892-6-CHEM

UF893-1-CHEM

UF893-1-CHEM

UF893-2-CHEM UF893-2-CHEM

UF893-5-CHEM

UF893-5-CHEM

UF893-6-CHEM

UF893-6-CHEM

UF894-1-CHEM UF894-1-CHEM

UF894-2-CHEM UF894-2-CHEM

UF894-5-CHEM

UF894-5-CHEM

UF894-6-CHEM

UF894-6-CHEM

UF895-1-CHEM

UF895-1-CHEM

UF895-2-CHEM UF895-2-CHEM

UF895-5-CHEM

UF895-5-CHEM

UF895-6-CHEM

UF895-6-CHEM

UF896-1-CHEM

UF896-1-CHEM

UF896-2-CHEM

UF896-2-CHEM

UF896-5-CHEM

UF896-5-CHEM

UF896-6-CHEM

UF896-6-CHEM

UF897-1-CHEM

UF897-1-CHEM

UF897-2-CHEM UF897-2-CHEM

UF897-5-CHEM

UF897-6-CHEM UF897-5-CHEM

UF897-6-CHEM UF898-1-CHEM

UF898-1-CHEM

UF898-2-CHEM

UF898-2-CHEM

UF898-5-CHEM

UF898-5-CHEM

UF898-6-CHEM UF898-6-CHEM

UF899-1-CHEM

UF899-1-CHEM

UF899-2-CHEM UF899-2-CHEM

UF899-5-CHEM

UF899-5-CHEM

UF899-6-CHEM UF899-6-CHEM

UF900-1-CHEM

UF900-1-CHEM

UF900-2-CHEM UF900-2-CHEM

UF900-5-CHEM

UF900-5-CHEM

UF900-6-CHEM UF900-6-CHEM

UF901-2-CHEM

UF901-2-CHEM

UF901-5-CHEM

UF901-5-CHEM

UF901-6-CHEM

UF901-6-CHEM

UF902-1-CHEM

UF902-1-CHEM

UF902-2-CHEM

UF902-2-CHEM

UF902-5-CHEM

UF902-6-CHEM UF902-5-CHEM

UF902-6-CHEM

UF904-1-CHEM

UF904-1-CHEM

UF904-2-CHEM

UF904-2-CHEM

UF904-5-CHEM

UF904-5-CHEM

UF904-6-CHEM

UF904-6-CHEM

UF905-1-CHEM

UF905-1-CHEM

UF905-2-CHEM UF905-2-CHEM

UF905-5-CHEM

UF905-5-CHEM

UF905-6-CHEM

UF905-6-CHEM

UF906-1-CHEM

UF906-1-CHEM

UF906-2-CHEM UF906-2-CHEM

UF906-5-CHEM

UF906-5-CHEM

UF906-6-CHEM UF906-6-CHEM

UF907-1-CHEM

UF907-1-CHEM

UF907-2-CHEM

UF907-2-CHEM

UF907-5-CHEM

UF907-5-CHEM

UF907-6-CHEM UF907-6-CHEM

UF908-1-CHEM

UF908-1-CHEM

UF908-2-CHEM

UF908-2-CHEM

UF908-5-CHEM

UF908-5-CHEM

UF908-6-CHEM UF908-6-CHEM

UF909-1-CHEM

UF909-1-CHEM

UF909-2-CHEM

UF909-2-CHEM

UF909-5-CHEM

UF909-5-CHEM

UF909-6-CHEM

UF909-6-CHEM

UF91-1-CHEM

UF91-1-CHEM

UF91-2-CHEM

UF91-2-CHEM

UF91-5-CHEM

UF91-5-CHEM

UF91-6-CHEM

UF91-6-CHEM

UF910-1-CHEM

UF910-1-CHEM

UF910-2-CHEM UF910-2-CHEM

UF910-5-CHEM

UF910-5-CHEM

UF910-6-CHEM

UF910-6-CHEM

UF911-1-CHEM

UF911-1-CHEM

UF911-2-CHEM

UF911-2-CHEM

UF911-5-CHEM

UF911-5-CHEM

UF911-6-CHEM

UF911-6-CHEM

UF912-1-CHEM UF912-1-CHEM

UF912-2-CHEM

UF912-2-CHEM

UF912-5-CHEM

UF912-5-CHEM

UF912-6-CHEM

UF912-6-CHEM

UF913-1-CHEM

UF913-1-CHEM

UF913-2-CHEM

UF913-2-CHEM

UF913-5-CHEM

UF913-5-CHEM

UF913-6-CHEM

UF913-6-CHEM

UF914-1-CHEM UF914-1-CHEM

UF914-2-CHEM UF914-2-CHEM

UF914-5-CHEM UF914-5-CHEM

UF914-6-CHEM

UF915-1-CHEM

UF915-1-CHEM

UF915-2-CHEM UF915-2-CHEM

UF915-5-CHEM

UF915-5-CHEM

UF915-6-CHEM

UF915-6-CHEM

UF916-1-CHEM

UF916-1-CHEM

UF916-2-CHEM

UF916-2-CHEM

UF916-5-CHEM

UF916-5-CHEM

UF916-6-CHEM UF916-6-CHEM

UF917-1-CHEM UF917-1-CHEM

UF917-2-CHEM

UF917-2-CHEM UF917-5-CHEM

UF917-5-CHEM

UF917-6-CHEM

UF917-6-CHEM

UF918-5-CHEM

UF918-5-CHEM

UF918-6-CHEM

UF918-6-CHEM UF919-1-CHEM UF919-1-CHEM

UF919-2-CHEM

UF919-2-CHEM

UF919-5-CHEM

UF919-5-CHEM

UF919-6-CHEM

UF919-6-CHEM

UF92-2-CHEM

UF92-2-CHEM

UF92-5-CHEM

UF92-5-CHEM

UF92-6-CHEM

UF92-6-CHEM

UF920-1-CHEM

UF920-1-CHEM

UF920-2-CHEM

UF920-2-CHEM

UF920-5-CHEM

UF920-5-CHEM

UF920-6-CHEM

UF920-6-CHEM

UF921-1-CHEM

UF921-1-CHEM

UF921-2-CHEM UF921-2-CHEM

UF921-5-CHEM

UF921-5-CHEM UF921-6-CHEM UF921-6-CHEM

UF922-1-CHEM

UF922-2-CHEM UF922-2-CHEM UF922-1-CHEM

UF922-5-CHEM

UF922-5-CHEM UF922-6-CHEM

UF922-6-CHEM

UF923-1-CHEM

UF923-1-CHEM

UF923-2-CHEM

UF923-5-CHEM

UF923-6-CHEM UF923-5-CHEM

UF923-6-CHEM

UF924-1-CHEM

UF924-1-CHEM UF924-1-CHEM UF924-2-CHEM

UF924-2-CHEM

UF924-5-CHEM

UF924-5-CHEM

UF924-6-CHEM

UF924-6-CHEM

UF925-1-CHEM

UF925-1-CHEM

UF925-2-CHEM UF925-2-CHEM

UF925-5-CHEM

UF925-5-CHEM

UF925-6-CHEM

UF925-6-CHEM

UF926-1-CHEM

UF926-1-CHEM

UF926-2-CHEM

UF926-2-CHEM

UF926-5-CHEM

UF926-5-CHEM

UF926-6-CHEM

UF926-6-CHEM

UF927-1-CHEM

UF927-1-CHEM

UF927-2-CHEM

UF927-2-CHEM UF927-5-CHEM

UF927-5-CHEM

UF927-6-CHEM UF927-6-CHEM

UF928-1-CHEM

UF928-1-CHEM

UF928-5-CHEM

UF928-5-CHEM

UF928-6-CHEM

UF928-6-CHEM

UF929-1-CHEM

UF929-1-CHEM

UF929-2-CHEM UF929-2-CHEM

UF929-5-CHEM

UF929-5-CHEM

UF929-6-CHEM UF929-6-CHEM

UF931-1-CHEM

UF931-1-CHEM

UF931-2-CHEM UF931-2-CHEM

UF931-5-CHEM

UF931-6-CHEM UF931-5-CHEM

UF931-6-CHEM

UF932-2-CHEM

UF932-2-CHEM

UF932-5-CHEM

UF932-5-CHEM

UF932-6-CHEM

UF932-6-CHEM

UF933-2-CHEM

UF933-2-CHEM

UF933-5-CHEM

UF933-5-CHEM

UF933-6-CHEM

UF934-1-CHEM

UF934-1-CHEM

UF934-2-CHEM

UF934-2-CHEM

UF934-5-CHEM

UF934-5-CHEM

UF934-6-CHEM UF934-6-CHEM

UF935-1-CHEM

UF935-1-CHEM

UF935-2-CHEM

UF935-2-CHEM

UF935-5-CHEM

UF935-5-CHEM

UF935-6-CHEM

UF935-6-CHEM UF936-1-CHEM

UF936-1-CHEM

UF936-2-CHEM

UF936-2-CHEM

UF936-5-CHEM

UF936-5-CHEM

UF936-6-CHEM

UF936-6-CHEM

UF937-1-CHEM

UF937-1-CHEM

UF937-2-CHEM

UF937-2-CHEM

UF937-5-CHEM

UF937-5-CHEM

UF937-6-CHEM UF937-6-CHEM

UF938-2-CHEM UF938-2-CHEM

UF938-5-CHEM

UF938-5-CHEM

UF938-6-CHEM UF938-6-CHEM

UF939-1-CHEM

UF939-1-CHEM

UF939-1-CHEM UF939-2-CHEM

UF939-2-CHEM

UF939-5-CHEM

UF939-5-CHEM

UF939-6-CHEM

UF939-6-CHEM

UF940-1-CHEM

UF940-1-CHEM

UF940-1-CHEM

UF940-2-CHEM UF940-2-CHEM

UF940-5-CHEM

UF940-5-CHEM

UF940-6-CHEM

UF940-6-CHEM

UF941-1-CHEM

UF941-1-CHEM

UF941-2-CHEM

UF941-2-CHEM

UF941-5-CHEM

UF941-5-CHEM

UF941-6-CHEM

UF941-6-CHEM

PC1

PC2 Scores

Sykes et al, Novaes et al

• HTP compositional analysis

– Glucan and xylan content

– Correlates with wet chemistry

• 24 samples in 4 hours- single reactor plate

– 2-3 days by wet chemistry

– Expandable to multiple plates

• Native or pretreated biomass

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

0.00 20.00 40.00 60.00 80.00

% Carbohydrate-HTP

% Carbohydrate-Wet Chemistry

prtx glucan prtx xylan

std glucan std xylan
Poplar Association Study-1st 2 Sets

• Error bars represent standard deviation of triplicate runs 

– reps equally distributed throughout plate; 24 samples per plate; 

– red bars are samples repeated in multiple plates

• All plate normalized to BESC poplar standard

– 8 standard runs per plate

• Data didn’t look very encouraging at first......
0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

16.0 18.0 20.0 22.0 24.0 26.0 28.0

sugar release - g/g biomass

% Lignin

Total Sugar

Glucose

Xylose

Poplar Association Study-755

•No apparent impact of lignin content on sugar release after 

pretreatment/enzyme hydrolysis

Selig et al
0.0

0.1

0.1

0.2

0.2

0.3

0.3

0.4

0.4

0.5

0.5

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5

sugar release - g/g biomass

S/G Lignins

Glucose

Xylose

Poplar Association Study- 755

•Glucan and xylan conversions show trend with S/G

•Xylan conversion is somewhat less scattered

Selig et al
0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

BES

...

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Combined Sugar Release (g/g biomass)

run #1

run #2

Poplar 4-CL Study- 100

• System consistently identifies outliers within small & large sample sets

Voelker et al
Danish Wheatstraw- 1130

• 1100 sample set – 25 separate experimental sets; strains, harvest 

conditions...etc

• Collaboration with Claus Felby, U. Copenhagen

• Consistent overlap between plates 

• Variation between sets and not plates

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0 240 480 720 960

Combined Sugar Release (g/g biomass)
Noble Foundation Switchgrass-58

• 58 samples included controls 

and replicates

• SSF carried out at ORNL

• Jonathon Mielenz lab

• First demonstration of 

fermentation/HTP pipeline 

correlation

0.000

0.100

0.200

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0.600

mg/mL sugar

Sugar Release - Pretreatment + Saccharification 

glucose xylose

200

220

240

260

280

300

320

340

360

0.150 0.200 0.250 0.300 0.350 0.400

mg ethanol/g dry biomass

recalcitrance screening glucose release in mg/g dry biomass

SSF vs. Recalcitrance Screening
Summary

• First high-throughput thermochemical pretreatment/enzyme digestion pipeline

– We have run dozens of campaigns 

– ~10,000 samples analyzed

• Solved numerous obstacles

– Biomass processing and dispensing

– Extractives and starch variability

– Pressure and evaporation control

– Rapid sugar analysis 

– HTP chemical analysis

• Severe pretreatment conditions possible

– High temperatures (150-200oC), pressures, low pH 

• HTP Analytical chemistry for lignin/carbohydrate content

– MBMS for total lignin, S/G ratios

– HTP compositional analysis for cellulose and xylan content

– Starch analysis is possible

• Data is comparative within sample sets

– Determine correlations of recalcitrance with environmental factors

– Measure effects of genetic variation

– Find outliers in large sample populations

• Capacity of 100s of samples/day

– Current bottleneck is sample preparation
Acknowledgments

• Stephen Decker

• Melvin Tucker

• Mike Himmel

• Rob Sykes

• Kristen Gracom

• Crissa Doeppke

• Melissa Carlile

• Cody Law

• Geoffrey Turner

• Mark Davis

• Charley Wyman

• Michael Studer

• Jaclyn DeMartini

• Heather McKenzie

This work was supported by the U.S. Department of Energy under Contract No. DE-AC36-08-GO28308 

with the National Renewable Energy Laboratory.

The BioEnergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the 

Office of Biological and Environmental Research in the DOE Office of Science.

• Martin Keller

• Brian Davison

• Gerry Tuskan

• Jonathon Mielenz

• Rick Nelson

• Fang Chen • William Everitt

• http://aspenmachining.com

• Claus Felby
References

• Somerville, C., S. Bauer, G. Brininstool, M. Facette, T. Hamann, J. Milne, E. Osborne, A. Paredez, S.

Persson, T. Raab, S. Vorwerk, H. Youngs, Toward a systems approach to understanding plant cell walls.

Science 306, 2206-2211 (2004).

• Sykes, R., Yung, M., Novaes, E., Kirst M., Peter,G., Davis, M., “High Throughput Screening of Plant Cell

Wall Composition Using Pyrolysis Molecular Beam Mass Spectroscopy” in Biofuels: Methods and

Protocols, Methods in Molecular Biology, 581, Mielenz, J.R.,(ed ) Humana Press pp 169-183 (2009).

• Novaes,E., Osorio,L., Drost, D.R., Miles, B.L., Boaventura-Novaes C.R.D.,Benedict,C., Dervinis, C., Yu, Q., 

Sykes R., Davis, M., Martin, T.A., Peter, G.F., Kirst, M., “Quantitative genetic analysis of biomass and 

wood chemistry of Populus under different nitrogen levels”, New Phytologist , 182 (4), 878-

890 (2009).

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A. Tuskan, L. Gunter, S. R. Decker, M. J. Selig, R. Sykes, M. E. Himmel, P. Kitin, O. Shevchenko, and S. H. 

Strauss. 2010 (in press). “Antisense down-regulation of 4CL expression alters lignification, tree growth 

and saccharification potential of field-grown poplar”. Plant Physiol.

• Selig, M. J., M. P. Tucker, R. W. Sykes, K. L. Reichel, R. Brunecky, M. E. Himmel, M. F. Davis, and S. R. 

Decker. (2010). “Biomass Recalcitrance Screening by Integrated High Throughput Hydrothermal 

Pretreatment and Enzymatic Saccharification”. Ind. Biotechnol. 6:104-111

• Decker, S. R., R. Brunecky, M. P. Tucker, M. E. Himmel, and M. J. Selig. (2009). “High Throughput 

Screening Techniques for Biomass Conversion”. Bioenergy Research. 2:79-192.