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.
// Actionable Intelligence – HPMS & On-board Analytics / Slide 3 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 4 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 5 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 6 June 2016
// 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
2015x07x21xx40 TOF MS ES+ BPI
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
2015x07x21xx30 TOF MS ES+ BPI
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
2015x07x21xx20 TOF MS ES+ BPI
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
2015x07x21xx10 TOF MS ES+ BPI
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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 7 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 8 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 9 June 2016
// 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
Data Courtesy of Mike Pacold of the Whitehead Institute0 .0 0 .5 1 .0 1 .5
0
1
2
3
[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 Time ~2 min
BGE 50% methanol, 2% formic acid
Q Exactive MS, Full MS scan
LOD/LOQ consistent with LC/MS
// Actionable Intelligence – HPMS & On-board Analytics / Slide 10 June 2016
// 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.
// Actionable Intelligence – HPMS & On-board Analytics / Slide 11 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 12 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 13 June 2016
// 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
50% methanol 2% formic acid
Sample Prep: • 1:10 dilution in 100 mM Am.Acet.
50% methanol 2% formic acid
• 0.22 µm filter
Growing application, ZipChip
provides excellent HT capabilities
// Actionable Intelligence – HPMS & On-board Analytics / Slide 14 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 15 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 16 June 2016
// 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
// Actionable Intelligence – HPMS & On-board Analytics / Slide 17 June 2016
//
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in Boston’s
Innovation
District.
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