EasySampler Tips, Tricks and Case Studies · PDF filePfizer Confidential │ 2 20 mL . Case Studies Guide Pfizer Confidential │ 3 Sampling Difficulty Scale Considerations: ... Case

EasySampler

Tips, Tricks and Case Studies

David Place

What is EasySampler?

• All-in-one sample collector AND

diluter/sample preparer

– Enables unattended pull sampling for offline

analysis (great for stability studies)

– Enables Quench-in-place operations

– Enables robust and precise sampling

from reaction Slurries

• Technology – Mechanical Actuation

– Sliding pocket inside a tube

– Carefully machined teflon tip/seal

Pfizer Confidential │ 2

20 mL

Case Studies Guide


Sampling Difficulty Scale Considerations:

• Reaction conditions (i.e. Temp, Pressure)

• Reaction texture

• Reaction mixture accessibility

Case Study Stats

Rxn Conc = 33 mg/mL

Temperature = 102 oC

EasySampler settings:

Commercial System

Q,D = 1:1 Acetonitrile/water

R = t-amyl alcohol

df = 160

Reactor

Reaction

SolventDiluentQuench Waste 10 mL

Sample Vial

ID = 0.8 mm

L = 0.15 m

V = 0.075 mL

Piston

Pump

200 uL Stroke

ID = 0.8 mm

L = 0.15+1.8 m

V = 1.0 mL

Needle

ID = 0.8 mm

L =0.1 m

V = 0.05 mL

ID = 0.8 mm

L = 0.8+1.8 m

V = 1.3 mL

Q D R

Dilution factor

Rule of Thumb for new chemistry:

For most Aromatic containing Materials monitored at 210-254 nm,

The following equation will give HPLC/UPLC results on scale

df = [Limiting reagent (mg)/ Total Volume (mL)]/ 0.3 mg/mL


Simple Dilution:

“I put one drop of the reaction mixture in 2 mL of diluent

then inject that solution”

Differentiation: Manual vs. EasySampler

Challenge: Try to sample a

solution manually 10 x. Can

you get the same result each

time?

Double Challenge: Now try to

do the same from a slurry.

What do you need to do to

ensure precise sampling?

Now which method do you feel

most confident in using if you

only get to sample once?


TRACER Material Sampling

Instrument

Sampling

Reproducibility

Naphthalenefull tip VWR

pippettor4%

Naphthalene EasySampler 5%

Naphthalene plastic pipette 6%

Naphthalene glass pipette 10%

Naphthalene syringe 46%

Acetaminophenfull tip VWR

pippettor22%

Acetaminophencut tip VWR

pippettor19%

Acetaminophen EasySampler 4%

Acetaminophen plastic pipette 52%

Acetaminophen glass pipette 61%

Acetaminophen syringe 37%

Common Sampling (Homogeneous)

Slurry Sampling (heterogeneous)

What does your reaction look like?

• Successfully sampled for kinetics in all of these cases w/ EasySampler to

provide modeling quality data sets


Thick Slurry Viscous w/ solids Undissolved

Inorganics

Tri-phasic

mixtures Just plain Ugly

(Cuprate salts)

O2

Oxygen sensitive

EasySampler = Representative Sampling

Representative sampling

• Sample containing all species,

solid/liquid/gas, contained

within a mixture which

represents the distributions of

those species in the bulk

media

• Examples:

– In Process Controls (IPC)

– Quality Control

– Kinetic Analysis

Selective Sampling

• Sample containing only a

portion of analyzable species

in a mixture due to physical

partitioning of the bulk media

• Examples:

– Mother Liquor analysis

– Solubility testing

– Phase splits

Slurries: Probe orientation is important

Test System in isopropanol:

• Dissolved tracer = Diphenyl ether

• Undissolved tracer = Caffeine

Best Tradeoff is pocket facing 180o

AWAY from agitator


Agi

90o

180o

270o

Reaction Mass

Pocket

0o

Case Ullman: Dealing with clogs (Kristin Wiglesworth)


Easy Hard *

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

100.00

0.0 5.0 10.0 15.0 20.0 25.0 30.0

UP

LC A

% a

t 2

20

nm

Time, h

Conversion: A% relative to Toluene (Int Std)

ArBr

ArOH

Product

Rxn Conc = ~125 mg/mL

Temperature = 85 oC


Alpha Prototype

Q,D = 1 v/v% water in DMSO

R = Toluene

df = 500

Reaction Matrix thickens

Preventing Clogs: Dissolution Rate important

• Lesson Learned: Choose the quench (Q) and diluent (D) solution wisely

• Use Quench operation in your EasySampler Method

– Quench oscillates the flow back and forth through the pocket allowing better mixing

and more time for solids to dissolve


0

2

4

6

8

10

12

14

16

0 2 4 6 8

Con

cent

rati

on

, mg/

mL

Time, minutes

Conc, mg/mL (50 % IPA)



Thin

ing

of S

lurr

y O

bser

ved

Dis

solu

tion

Ach

ieve

d

kLA = 2.2 ± 1.9% sec-1

Test system: Caffeine in IPA/water Mixtures

Empirical observation:

If at least 90% of the solid

Does not dissolve by 4 min

Clogging or inconsistent

Sampling will result.

Not recommended to use

Dilution factors less than 160

In commercial systems

Case Imidazolide : Quench-In-Place (Shu Yu)


Easy Hard *

Rxn Conc = 100 mg/mL

Temperature = 22 – 50 oC


Beta Prototype

Q,D = 1:1 DMSO/MeOH

R = THF

df = 330

Dete

rmin

e A

ctivation E

nerg

ies

Slurries – Things to Watch for

Sampling from Slurry depends

on Agitation rate and

Probe Position

Stratification

Solid

crust

EasySampler Facilitates Workflow

• EasySampler is an enabling technology that facilitates integration

between hardware and software assets at Pfizer. Integration leads to

stronger fundamental and dynamic models of chemical processes.


Without a technology like this

All other steps become

More tedious and difficult

Case C-H Activation: Trending and IPC specification

(Brian Vanderplas)

Rxn Conc = 33 mg/mL

Temperature = 102 oC


Commercial System


R = t-amyl alcohol

df = 160


7 h

IPC Target 5 A% Substrate

Easy Hard *

Challenge: Calling reaction endpoint

Benefit: Further mechanistic insights

Case De-Boc: Dynochem Modeling (Marshall Greenwood, Brian Vanderplas)


Easy Hard *

Rxn Conc = 250190 mg/mL

Temperature = 60 oC


Commercial System


R = Methanol

df = 350

Base-lining kinetics sets stage for future

mechanistic understanding

Challenge: catalytic reaction showing

Dependence on catalyst why?

k1= 14 x 10-4 1/s, C.I. 11%

k2 = 9.6 x 10-4 1/s, C.I. 11%

Case Amination: Virtual Design Space (Jeff Sperry)

Best Trade-off:

T = 27 oC, NH4OH = 5.9 mol equiv.


Easy Hard *

Case Pressure Reactors (Teresa Makowski, Jason Mustakis)

• Mock System using JM

Carbon support (~1 wt% vs.

reaction mass)

• Slow leaks observed when

probe is actuated

• Consistent Sampling

observed up to 75 psig

• Heterogeneous catalyst

actually benefit stopping

leaks


Pressure, psig Carbon TRACER %RSD Sample Weight %RSD Accuracy, % deviation

0.10 no 0.68% 0.62% +5.4%

75 no 0.70% 0.76% +181%

75 yes 2.05% 0.99% +6.1%

Disclaimer: This is Out-of-spec use of the EasySampler but…shows promise

• Homogeneous Catalyst system

• Seeing solvent dependent

degradation of chiral center

• EasySampler provided a

method to sample without

breaking pressure

Case Pd Catalysed Carbonylation (Teresa Makowski)


Easy Hard *

Rxn Conc ~ 116 mg/mL

Pressure, Temperature = 50 psi, 80 oC


Commercial System

Q,D = Acetonitrile; R = Methanol

df = 160

Case Heterogenous Catalysis (Teresa Makowski)


Easy Hard *

Rxn Conc ~ 50 mg/mL

Pressure, Temperature = 70 psi, 70 oC


Commercial System

Q,D = Methanol; R = Methanol

df = 160

Rxn Conc ~ 50 mg/mL

Pressure, Temperature = 25-50 psi, 70 oC


Commercial System

Q,D = Methanol; R = Methanol

df = 160

Seeing evidence of pump/clog issues

PUMP ERROR: Pressure testing Reactors with

EasySampler installed

• Most probe technologies allow pressure testing with an empty reactor

• Not recommended for EasySampler, fill with solvent to avoid EasySampler

Pump errors Gas leaks cause complete evacuation of lines over time


Take-Away #1

• Two key features of the reaction mixture should be

present in order to acquire Representative samples:

1.Reaction mixture is flowable/suspenable

2.Reaction mixture is dissolvable

• When Mixture doesn’t have these properties, selective

sampling is achieved and results obtained will be skewed

to favor observation of certain reaction components over

others


Take Away #2

THINGS TO CONSIDER BEFORE SAMPLING for UPLC/HPLC

These concepts are important regardless of the equipment used:

1.Can I dissolve the mixture (and with what)?

2.What dilution factor should I use?

3.Where should I sample the mixture?

4.Can I repeat and get the same result?

5.Easysampler: is the probe pocket oriented 180o away from the agitator

For most Aromatic containing Materials monitored at 210-254 nm,

The following equation will give HPLC/UPLC results on scale

df = [Limiting reagent (mg)/ Total Volume (mL)]/ 0.3 mg/mL


Acknowledgements

• Pfizer

Joel Hawkins

Stephanie Dolph

George Reid

Hahdi Perfect

Barb Sitter

Daniel Price

• Mettler Toledo

Peter Blacklin

Vaso Vlachos

Reni Mueller

Cis Vandereycken

Roel Ferwerda

Erich Schellinger

Nilesh Shah

• Development Partners

Luc Moens (JNJ)

Tom Maes (JNJ)

Chris Nichols (JNJ)

Antonio Ramirez (BMS)

Selin Aytar (BMS)

Jon Jurica (Merck)

Charles Orella (Merck)

Mats Ridemark (AZ)

Christian Bernlind (AZ)


Back-up Slides


Slurry vs. Temp

Control

@ 25 °C

60 °C to

reflux

10 °C +

1 day

Slurry

thins Recryst

Solid

crust

5/11/2015

Site Install Performance Metrics


Leak Detection Metrics Sampling Performance Metrics

SN# Pump SN# Probe Back Pressure, barg Tube ID, mm DYE TEST Pump performance Sampling %RSD Pocket Volume, uL Real Stroke, mL

Criteria For Success --> 1.7 +/- 0.3 0.80 +/- 0.16 < 1% < 5% 20 +/- 5 0.2000 +/- 0.0060

B432897873 B438042793 1.64 0.81 PASS 0.13% 5.3% 29.95 0.1983

B443190018 B443190028 1.63 0.81 PASS 0.39% 3.3% 20.60 0.1977

B432897872 B438042794 1.52 0.83 PASS 0.50% 3.2% 20.2 0.1983

B435975955 B443190040 1.64 0.81 PASS 0.49% 5.0% 22.2 0.2000

B435975954 B441110195 1.63 0.81 PASS 0.31% 2.4% 22.0 0.1974

B435975956 B441110190 1.50 0.83 PASS 0.52% 1.5% 21.8 0.1973

B435975962 B441110192 1.73 0.80 PASS 0.14% 4.2% 21.3 0.1992

B443190019 B443190029 1.76 0.80 PASS 0.09% 4.3% 20.93 0.1992

Ismatec MODEL

78017

Isopropanol

Solvent Resrevoir

P

Alpha Pfastrack

Pressure Tranducer

1/8" PTFE

1/8" PTFE

PTFE

ID=0.8 mm

L= 1.8 m

PTFE

ID=0.8 mm

L= 1.8 m

Displayed Text Description Manufacturer Material ModelIsmatec MODEL

78017 Peristaltic pump Ismatec

Isopropanol

Solvent ResrevoirVented Schott bottle

Equipment List Toluene tracer in IPA

Leak Detection Metrics

Test # Notes SN# Probe Back Pressure, barg Tube ID, mm DYE TEST

1 Install B438042794 2.99 0.70 PASS

2 Changed Tubing B438042794 3.20 0.69 PASS

3 Changed Tip B438042794 2.15 0.76 PASS

4 Changed Head B438042794 1.52 0.83 PASS

Hagen-Poisieulle used to Calculate Tube ID

What to expect when the Teflon Tip dies

Checklist:

1. Was the sample tip completely

immersed?

2. For slurries, Probe optimally

oriented in the flow?

3. Does diluent dissolve solids?

4. UPLC/HLC integrator correctly

identifying peaks in each

sample?


Prim

ary

Convers

ion, A

%

Impurities, A

%

Change Teflon Tip out with new one

Time, h

Observed Teflon Tip lifetimes (so far)

• Running at 102 oC and/or 50 psi + for 12-24 h = 400-450 actuations ( ~ 14 expts )

• HOWEVER, one system up to 770 actuations and still functioning optimally

Cross Contamination

Scenario: Q = Water, D= MeCN, R = THF; 1 g scale

• Assume 100 ppm water contaminates reaction solvent ( R ) in the 20

uL pocket.

• How many actuations are required to add 1% of the water required to

completely quench reaction mixture?


610

Tracer Study (Zero Series)

Tracer = Diphenyl ether

(DPE)

• Sample Reaction

mixture 10 x

• Collect Samples at

max df = 450

• Sample reaction pot

manually


Reactor

Reaction

SolventDiluentQuench Waste 10 mL

Sample Vial

ID = 0.8 mm

L = 0.15 m

V = 0.075 mL

Piston

Pump

200 uL Stroke

ID = 0.8 mm

L = 0.15+1.8 m

V = 1.0 mL

Needle

ID = 0.8 mm

L =0.1 m

V = 0.05 mL

ID = 0.8 mm

L = 0.8+1.8 m

V = 1.3 mL

IPA D IPA

25.4 wt% DPE

In IPA 8.9 mg/g Toluene

In IPA

(4 mL)

Result:

DPE in Reaction pot = 71 ppm

Diluent Contamination = 279 ppm

Documents

EasySampler Tips, Tricks and Case Studies · PDF filePfizer Confidential │ 2 20 mL . Case Studies Guide Pfizer Confidential │ 3 Sampling Difficulty Scale Considerations: ... Case