Page 84 - Application Notebook - Solution for Food Safety

Application No.C141
News

Table 1 Analytical Conditions

System : Nexera X2 System : LCMS-8060
Column : Shim-pack XR-ODS ᶘ Ionization : Heated ESI
(50 mm L. × 2 mm I.D., 1.6 μm) Probe Voltage : +1 kV (positive ionization)
Column Temperature : 40 °C Temperature : Interface: 250 °C
Mobile Phases : A: Water + 0.1 % formic acid Desolvation Line: 150 °C
B: Acetonitrile Heater Block: 200 °C
Flowrate : 500 μL/min Gas Flow : Nebulizing Gas: 3 L/min
Gradient : 2 %B (0.00 min) > 25 %B (7.00 min) > Heating Gas: 20 L/min
95 %B (7.10-8.00 min) > 2 %B (8.10-10.00 min) Drying Gas: 5 L/min
Injection Volume : 10 μL

Table 2 MS/MS Acquisition Parameters
MRM Transitions Name Polarity Quan Qual1 Qual2
EGEQEWGTPGSEVR + 780.85 > 802.40 780.85 > 644.35 780.85 > 316.10
NNPFYFPSR + 571.25 > 669.35 571.25 > 506.25 571.25 > 229.10
IPSGFISYILNR + 690.40 > 765.45 690.40 > 211.15 690.40 > 502.25
SSDNEGVIVK + 524.25 > 515.35 524.25 > 359.25 524.25 > 175.05
GSEEEDITNPINLR + 793.90 > 726.45 793.90 > 612.40 793.90 > 402.25
GTGNLELVAVR + 564.80 > 686.40 564.80 > 557.40 564.80 > 444.30
EGEQEWGTPGSHVR + 784.85 > 652.35 784.85 > 555.30 784.85 > 316.10
SSENNEGVIVK + 588.30 > 515.35 588.30 > 359.25 588.30 > 246.20
GSEEEGDITNPINLR + 822.40 > 726.45 822.40 > 612.40 822.40 > 402.25

Dwell Time : 41 to 130 msec depending upon the number of concomitant transitions to ensure to have at
least 15 points per peak (max total loop time 400 msec).
Pause Time : 3 msec
CID Pressure : 300 kPa
Quadrupole Resolution : Q1: Unit Q3: Unit

Q Interface Optimization Q Effect of Surfactant During Digestion
Ionization parameters optimization was performed A higher intensity of peptides by addition of a
using companion software ISSS (Interface Setting surfactant during tryptic digestion was expected due to
Support Software, Shimadzu Corp.). As a result, improved digestion efficiency. However, the intensity of
sensitivity was improved more than twofold compared peptides were relatively worse by adding surfactant.
to default values. Thus, no surfactant was used for tryptic digestion.

Intensity of GTG... 500000 Intensity of GTG... 300000 0 1.0 (×1,000,000) 1.0 (×1,000,000)
400000
200000
300000
200000
100000
100000
60.7 %
0
250
200
4
0
2
Interface voltage (KV) 6 100 150 DL temp (degC) 300 350
0.5 0.5
250000
250000
Intensity of GTG... 200000 0 Intensity of GTG... 200000 0 0.0 0.0
150000
150000
100000
100000
50000
50000
450
350
250
20
15
0
5
10
Drying gas flow (L/min) 25 200000 150 CID gas pressure (kPa) 550 4.0 w/o surfactant 4.4 4.0 w/ surfactant 4.4
300000
Intensity of GTG... 200000 0 Intensity of GTG... 150000 0 Fig. 5 Difference of the Chromatograms of Peptide GTG…
100000
100000
50000
by Addition of Surfactant
0
3
2
20
10
15
1
5
Heating gas flow (L/min) 25 0 Nebulizing gas flow (L/min) 4
Fig. 4 Interface Optimization Results