Microfluidic CE-ESI Small Molecule CE-ESI-MS …908devices.com/wp-content/uploads/2016/02/ZipChip-CE-ESI...// Actionable Intelligence –HPMS & On-board Analytics / Slide 3 June 2016

June 2016// Actionable Intelligence – HPMS & On-board Analytics / Slide 1

ZipChipTM

Microfluidic CE-ESI

Small Molecule CE-ESI-MS Applications

Please visit us at

http://www.908devices.com/

// Actionable Intelligence – HPMS & On-board Analytics / Slide 2 June 2016

// ZipChip CE-ESI Amino Acids/Metabolites Overview

• Fast, high-resolution CE separations under 3 min for amino acids/metabolites in matrices of

varying complexity. ZipChip CE-ESI MS quantitative with LOQs comparable to LC/MS.

• Corresponding LC elution significantly longer and method development is more challenging.

Conventional CE systems have much longer CE migration times and lower resolution.

• Minimal sample preparation for complex matrices courtesy of on-chip desalting

• Low volume nl injections advantageous for precious samples. Negligible sample to sample

carryover in ZipChip CE-ESI. Optimized CE method available for metabolomics samples.

• Can increase run time and number of MS and MS/MS scans across peak by switching from the

ZipChip HS Chip (10 cm) to ZipChip HR (22 cm). Alternately, analysis time can be increased by

changing BGE composition or CE voltage differential.

• Extending analysis run time can also allow for higher resolution Orbitrap MS, MS/MS scanning.

• Autosampler capability near completion for high throughput SM assays. Anionic CE separation

capability to follow.


// ZipChip CE-ESI MS of Amino Acids

2.5 uM Promega Complete AA MixZipChip HS – Thermo Fisher Exactive Plus EMR Orbitrap MS

BGE: 50% methanol, 2% formic acid (pH 2.2)

1.8

min

Very Fast High Resolution AA Separations


// ZipChip CE-ESI versus Conventional CE for Amino Acids

ZipChip Microfluidic CE-ESI

1.8

min

Conventional Capilliary CE(Beckman CE-8000)

24

min

Much Slower CE Migration Times

Lower Resolution Separations


// ZipChip CE-ESI MS of Amino Acids

Isoleucine Leucine

FWHM = 0.55 secondsThermo Exactive Plus EMR Orbitrap MS

Resolution: 17,500

1 microscan

Max IT: 50 ms

70-1000 m/z

Many data points across each peak for

qualitative and quantitative MS and MS/MS


// ZipChip CE-ESI - Migration Time Reproducibility (AA Mix)

10 cm CEESI_PEG allover_10 uM AA mix_external trigger test

Time

0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 2.05

%

0

100

0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 2.05

%

0

100

0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 2.05

%

0

100

0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 2.05

%

0

100

0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 2.05

%

0

100

2015x07x21xx50 TOF MS ES+ BPI

7.68e41.04

175.119

0.99147.112

1.08156.078

1.78166.0871.68

150.0611.55132.104

1.51132.104

1.76116.071

1.82205.100 1.98

182.080

1.86148.063


7.68e41.02175.119

0.97147.112

1.06156.078

1.75166.0871.65

150.0611.49132.104

1.46118.088

1.52132.104 1.72

116.071

1.79205.100 1.94

182.080

1.83148.063


7.68e41.01175.119

0.96147.112

1.05156.078

1.72166.0871.62

150.0611.47

132.1041.43

118.088

1.50132.104

1.68116.071

1.76205.100 1.90

182.080

1.79148.063


7.68e41.01

175.119

0.96147.112

1.06156.078

1.73166.0871.63

150.0611.47132.104

1.44118.088

1.51132.104

1.78205.100 1.92

182.080

1.80148.063


7.68e41.00

175.119

0.95147.112

1.05156.078

1.71166.0871.61

150.0611.49132.104

1.46132.104 1.66

116.071

1.76205.100

1.90182.080

1.78148.063

lysine isoleucine leucine aspartic

acid

Run #10

Run #20

Run #30

Run #40

Run #50


// ZipChip CE-ESI Migration Time Reproducibility

0.7

0.9

1.1

1.3

1.5

1.7

1.9

2.1

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50

Mig

rati

on

Tim

e (m

inu

tes)

lysine

isoleucine

leucine

aspartic acid

1.1% RSD

1.2% RSD

1.2% RSD

1.6% RSD


// Quantitative ZipChip CE-ESI Analysis of Amino Acids

0 1 0 2 0 3 0 4 0

0

2 0

4 0

6 0

8 0

[m e ta b o lite ] (µ M )

Are

a R

ati

o (

me

tab

oli

te p

ea

k/i

nte

rna

l s

tan

da

rd)

M e ta b o lite S ta n d a rd C u rv e s ly s in e

o rn ith in e

a rg in in e

h is t id in e

g ly c in e

c r e a t in e

c a r n it in e

a la n in e

a c e ty lc a r n it in e

v a lin e

is o le u c in e

le u c in e

s e r in e

th re o n in e

p h e n y la la n in e

p ro lin e

g lu ta m in e

a c e ty ls e r in e

g lu ta m a te

ty r o s in e

b e ta in e

h y d ro x y p ro lin e

m e th io n in e

t r y p to p h a n

c it r u ll in e

c y s te in e

a s p a ra g in e

a s p a r ta te

c y s t in e

ZipChip HS, Analysis ~2 min

50% methanol 2% formic acid

Q Exactive MS, Full scan

Data Courtesy of Mike Pacold of the Whitehead Institute


// Quantitative ZipChip CE-ESI Analysis of Amino Acids

0 1 0 2 0 3 0 4 0

0

2 0

4 0

6 0

8 0


Are

a R

ati

o (

me

tab

oli

te p

ea

k/i

nte

rna

l s

tan

da

rd)


o rn ith in e

a rg in in e

h is t id in e

g ly c in e

c r e a t in e

c a r n it in e

a la n in e


v a lin e

is o le u c in e

le u c in e

s e r in e

th re o n in e


p ro lin e

g lu ta m in e


g lu ta m a te

ty r o s in e

b e ta in e


m e th io n in e

t r y p to p h a n

c it r u ll in e

c y s te in e

a s p a ra g in e

a s p a r ta te

c y s t in e

Data Courtesy of Mike Pacold of the Whitehead Institute0 .0 0 .5 1 .0 1 .5

0

1

2

3


Are

a R

ati

o (

me

tab

oli

te p

ea

k/i

nte

rna

l s

tan

da

rd)


o rn ith in e

a rg in in e

h is t id in e

g ly c in e

c r e a t in e

c a r n it in e

a la n in e


v a lin e

is o le u c in e

le u c in e

s e r in e

th re o n in e


p ro lin e

g lu ta m in e


g lu ta m a te

ty r o s in e

b e ta in e


m e th io n in e

t r y p to p h a n

c it r u ll in e

c y s te in e

a s p a ra g in e

a s p a r ta te

c y s t in e

ZipChip HS, Analysis Time ~2 min

BGE 50% methanol, 2% formic acid

Q Exactive MS, Full MS scan

LOD/LOQ consistent with LC/MS


// ZipChip CE-ESI MS of Cell Extract (Metabolomics)

MDA-MB-231 Breast Cancer Cells

Washed with 150 mM ammonium chloride

Lysed/extracted with 80:20 methanol:water

Filtered (0.45 mm nylon SpinX)

ZipChip HS (10 cm channel)

Full Scan MS (50-1000 m/z)

No Additional Sample Preparation

Less than 3 min analysis. Perfect for

the running of high-throughput metabolomics

assays.


// ZipChip CE-ESI-MS of Mouse Plasma

Mouse Plasma Sample

Extracted with 80:20 methanol:water

Diluted with BGE + 100 mM ammonium bicarbonate

(50x total dilution)

ZipChip HS (10 cm), Full Scan MS (50-1000 m/z)

No Additional Sample Preparation

Less than 2 min analysis time. Perfect for

high-throughput metabolomics assays


// ZipChip CE-ESI MS of Urine

12

Urine Sample

Diluted with BGE: 50% methanol, 2% formic acid (pH 2.2)

(10x total dilution)

Filtered (0.45 mm nylon SpinX)

ZipChip HS – Full Scan MS (50-1000 m/z)

No Additional Sample Preparation, Analysis time ~2 min

Some prominent peaks labeled

nL sample injections, volatile BGE, nanoflow ESI

- all aid robust MS performance


// ZipChip CE-ESI of Cell Culture Media – Metabolite Consumption

E. Coli (MG1655)

S. Thermophilus (BAA-250)

M17 Broth Before Growth

ZipChip HS – high speed analysis


Sample Prep: • 1:10 dilution in 100 mM Am.Acet.


• 0.22 µm filter

Growing application, ZipChip

provides excellent HT capabilities


// ZipChip Autosampler Overview

14

Autosampler

ZipChip Interface AS

transfer line

Thermo Xcalibur

Mass Spec

The autosampler is acting as an auto-pipetter,

delivering rinse BGE, or samples from well-

plates. Injections, separation control are

managed from the ZipChip Interface;

Thermo AS platform


// ZipChip Autosampler Overview

15

AutosamplerZipChip Interface AStransfer line

~ 50 uL internal volume

1 m @ 250 um ID

waste

ZC sample

well

BGE (rinse,

consumption

approx. 0.5

mL per

sample)

96 well plate

20 uL

minimum

(sample &

rinsate)

vacuumZCI contains

stock of BGE

internal for non-

sample well

supply


// ZipChip Autosampler - Anticipated Timing Diagram

16

Rinse (20 sec):

Evacuate sample in reservoir (2 sec)

500 uL rinse with BGE (15 sec)

CE-MS (2-3 min):

Deliver Sample (10 sec):

5 uL minimum in microwell plate


//

908 Devices is

headquartered

in Boston’s

Innovation

District.

Thank You

Documents

Microfluidic CE-ESI Small Molecule CE-ESI-MS …908devices.com/wp-content/uploads/2016/02/ZipChip-CE-ESI...// Actionable Intelligence –HPMS & On-board Analytics / Slide 3 June 2016