AI-64310-CRFT

Thermo Scientific Poster Note

•

PN-64103-ASMS-EN-0614S

FIGURE 6. Versatility chart for Tran

quadrupole-Orbitrap MS platform.

tive™ hybrid quadrupole-

heated electrospray

ion used to quantitate results.

FIGURE 4. Upper trace: 384 sulfatide sample injections collected into a single

data file, 9 s data window. Lower trace: Scaling to 1 min (four injections).

E:\Q-ExactiveData\...\Sample1537

11/21/20133:17:55PM

Sample1537

N/C

RT:

0.00-107.25

SM:

itive (250-700

m/z

) and

,000. A 30 s MS data window,

as used.

ll scan negative ionization

100

nce

100

RelativeAbundance

RT:50.96

AA:110520

SN:363

RT:63.28

AA:108578

SN:348

RT:91.24

AA: 96646

SN: 329

RT:66.64

AA: 94929

SN: 325

RT:87.88

AA:93785

SN:324

RT:0.68

AA:102558

SN:319

RT:75.59

AA:93956

SN:318

RT:9.64

AA:96896

SN:314

RT:95.19

AA:92747

SN:311

RT:23.56

AA:91786

SN:306

RT:44.26

AA:92712

SN:306

RT:101.37

AA:90521

SN:305

RT:16.34

AA: 95527

SN: 302

RT:26.92

AA: 91004

SN:302

RT:37.58

AA:88315

SN:291

RT:50.97

AA:703842

SN:1949

RT:58.81

AA:674847

SN:1875

RT:63.29

AA:695572

SN:1870

RT:0.68

AA:665874

SN:1756

RT:87.89

AA:603052

SN:1741

RT: 75.60

AA:608291

SN:1697

RT:9.64

AA:622674

SN:1686

RT:16.35

AA:634708

SN:1674

RT:26.35

AA:583436

SN:1661

RT:93.48

AA:589245

SN:1651

RT:44.26

AA: 597256

SN: 1650

RT:101.37

AA: 576524

SN: 1640

RT: 31.40

AA:592022

SN:1619

NL:

9.95E4

m/z=

862.59973-

862.61699

MS ICIS

Sample1537

NL:

6.13E5

m/z=

888.61512-

888.63290

MS ICIS

6,000. Two acquisition styles

ed. The first style of

s data windows) involved a

tographic separation of the

ata not shown). A two-tiered

d i

l d fil

100 105

Time (min)

RelativeAbunda

Sample1537

RT:

53.13 - 54.13

SM:

100

RT:53.21

AA:96067

SN:311

RT:53.82

AA:78593

SN:259

RT:54.07

AA: 80310

NL:

8.52E4

m/z=

862.59973-

86261699

cte nto a s ng e ata e

hroughput tier demonstrated

mpounds based solely on

100

tiveAbundance

RelativeAbundan

SN:236

RT:53.52

AA:69825

SN:151

RT:53.16

AA:9375

SN: 31

RT:54.02

AA: 9340

SN: 28

RT:53.77

AA:9827

SN:28

RT:53.46

AA:7692

SN: 17

RT:53.21

AA:605017

SN:1626

RT:53.82

AA:494857

SN:1347

RT:54.07

AA:523323

SN:1297

RT:53.52

AA:448931

SN:829

MS ICIS

Sample1537

NL:

5.11E5

m/z=

888.61512-

888.63290

MS ICIS

Sample1537

Results

C St d #1

med using Thermo

, individually collected data

e version 3.0. Post-

53.2

53.3

53.4

53.5

53.6

53.7

53.8

53.9

54.0

54.1

Time (min)

Rela

ase u y :

Using a 100 s LC/MS research method with sample cleanup and chromatographic

resolution, antiretrovirals were acquired within a data window of 30 s using a

multiplexed (two-channel) LC system. Within the dynamic range investigated

all four

standard curve matrices gave linearity between R

= 0.96 and 0.99. Internal standard

cted into a single file was

rmo Scientific™ QuickCalc

ross four matrix preparations

Conclusion

%RSD across two channels was maintained at

or better than

16% for each curve.

Case Study #2:

The first workflow (1000 separate data files using 21 s data windows) resulted in the

acquisition of an average of 1.7 samples per minute with baseline chromatographic

.25

2978793

: 0.29

: 83570949

0.24

NL: 4.78E6

m/z=

267.12326-267.12594

F: FTMS + p ESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL: 1.74E7

m/z=

671.38819-671.39491

F: FTMS + p ESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL: 3.82E3

RT:

0.00 - 0.52

SM:

100

RelativeAbundance

100

RelativeAbundance

RT: 0.25

AA: 19856397

RT: 0.29

AA: 48388013

RT: 0.26

NL: 4.15E6

m/z=

267.12326-267.12594

F: FTMS + pESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL: 9.82E6

m/z=

671.38819-671.39491

F: FTMS + pESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL: 3.29E3



This work has effectively demon

with Q Exactive MS platform (Fi

online sample cleanup as neede

multiple injections into a single d

throughput, whether for researc

resolution of three analytes. The fourth analyte, co-eluting with another sulfatide, was

earmarked as the internal standard.

The second workflow (three sets of 384 injections collected into 3 single data files

using 21 s data windows) resulted in the acquisition of an average of 2.2 samples per

minute with peaks chromatographically resolved.

16730

RT: 0.36

AA: 3235427

T: 0.31

A: 51351138

T 034

m/z=

266.08815-266.09081

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004

NL: 6.82E5

m/z=

314.01854-314.02168

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004

NL: 1.19E7

m/z=

309.11059-309.11369

F: FTMS + p ESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL:316E7

100

RelativeAbundance

100

RelativeAbundance

100

RelativeAbundance

AA: 15227

RT: 0.36

AA: 3068603

RT: 0.31

AA: 64432405

RT 034

m/z=

266.08815-266.09081

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004

NL: 6.49E5

m/z=

314.01854-314.02168

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004

NL: 1.46E7

m/z=

309.11059-309.11369

F: FTMS + pESI Full

ms [250.00-700.00]

MS ICIS

StdA_250_004

NL:386E7

chromatography. The use of hig

in both the sensitivity and selecti

capabilities of the multi-channel

platform can be used to continu

compounds are eluting. QuickC

those multiple injections into a si

The final workflow (one set of 384 injections into a single file using 9 s data windows)

resulted in the acquisition of 4.0 samples per minute with resolution by accurate mass

in an ultra-high-throughput workflow. All workflows are summarized in Table 3.

Robustness of the assay was demonstrated with a combined %RSD of multiple

Crashed

asma

ACN-Crashed

Whole Blood

0.4

e (min)

: .

A: 175642230

m/z=

307.09604-307.09912

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004

0.0

0.2

0.4

Time (min)

100

RelativeAbundance

: .

AA: 215104504

RT: 0.13

AA: 98765

m/z=

307.09604-307.09912

F: FTMS -p ESI Full

ms [250.00-350.00]

MS ICIS

StdA_250_004



In conclusion, it is important to n

determinations must involve con

such as chromatographic separ

interference, and carryover ass

TABLE 3. Overview of acquisition styles for data collection of 384-sample batches

th CTC D l A DLW AS ith th f

h l T

d II LX 4 t

injections (1152# multiplexed) for the 21 s data window workflow of ≤5.8% and with

%RSDs of multiple injections (384# multiplexed) for the 9 s data window workflow of

≤2.8% .

ections collected into a

to 1.75 min (four injections).

autosampler. Each of these con

throughput. Overall, the greates

that it can bring to either clinical

us ng e

ua - rm

w e our-c anne ranscen - sys em

running cross-channel staggered injections to the Q Exactive HRAM MS.

Data

Window

LCMethod

Length

Injections

RT:155.08

AA:25839

SN:254

RT:162.44

AA:25516

SN:253

RT:132.99

AA:24979

SN:248

RT:147.72

AA:26397

SN:247

RT:143.23

AA:80268

SN:384

RT:163.47

AA:79306

SN:382

RT:150.59

AA:77699

SN:372

.14

RT:137.72

AA:75846

SN:357

NL: 3.54E4

m/z=

806.54173-

806.54979 MS

ICISSample001

NL: 9.64E4

m/z=

862.60405-

862.61267 MS

ICISSample001

NL:239E5

References

1. Chassaing, C.; Robinson, S.;

20-24.

AcquisitionStyle

Length

(s)

(min)

Chromatography

perminute

StandardLC-MS

w/384datafiles

N/A

1.6

Baselineseparationof3

analytes

0.63

130

140

150

160

170

RT:143.31

AA:222421

SN:356

RT:167.23

AA:219008

SN:341

RT:158.04

AA:220406

SN:341

RT:130.44

AA:217567

SN:337

RT:143.23

AA:515626

SN:1635

RT:176.36

AA:499585

SN:1620

RT:163.47

AA:503520

SN:1603

RT:137.72

AA:492410

SN:1567

m/z=

890.63521-

890.64411 MS

ICISSample001

NL: 6.05E5

m/z=

888.61957-

888.62845 MS

ICISSample001

TranscendIILX-4system

with384datafiles

1.6

Baselineseparationof3

analytes

1.7

TranscendIILX-4system

with1datafile

(DT Submit)

1.6

Baselineseparationof3

analytes

2.2

RT:104.45

AA:19941

SN:239

RT:104.59

AA:75250

SN:335

RT:104.54

AA:8014

SN:40

RT:104.68

AA:217095

SN:310

RT:104.62

AA:96459

SN 151

NL: 2.67E4

m/z=

806.54173-

806.54979 MS

ICISSample001

NL: 7.35E4

m/z=

862.60405-

862.61267 MS

ICISSample001

NL: 1.82E5

m/z=

890.63521-

890.64411 MS

CTC is a trademark of CTC Analytics AG, Zwing

Fisher Scientific and its subsidiaries.

TranscendIILX-4system

with1datafile

(DT_Submit)

1.0

Co-elutionof2analytes

4.0

104.3

104.4

104.5

104.6

104.7

104.8

RT:104.59

AA:493233

SN:1473

ICISSample001

NL: 4.62E5

m/z=

888.61957-

888.62845 MS

ICISSample001

For Research use only. Not for u

This information is not intended to encourage us

intellectual property rights of others.