Page 10 - Shimadzu LCMS-8060

High-Sensitivity Peptide Detection

High sensitivity and fast scanning capabilities has opened up new workflows for quantitative proteomics.

In the detection of the tryptic peptide AFVFPK (from C-reactive protein, CRP) as a biomarker for inflammation 3 mg/L is reported as an
average level of CRP in plasma. The LCMS-8060 delivers a highly sensitive detection of AFVFPK with the lowest calibration curve point at
0.008 mg/L.
Expand your capabilities in quantitation 200 AFVFPK (isotope labeled peptide)
0.00805–322 mg/L in plasma
Ultra sensitive peptide analysis 100 (%) 150 Y = 0.49579X + 0.00519
r 2 = 0.998
5 attomol on column Synthetic peptide area ratio (heavy / light) 100 15 levels
AFVFPK
and lipid profiling 50 in neat solution 50
blank solvent
0
Advanced design, fast workflows and added business value 0 0 100 200 300 400 mg/L
2.00 2.25 2.50 2.75 3.00 3.25 3.50 min
help the LCMS-8060 deliver on a broad range of applications. (%) Spiked conc. Calculated conc. Accuracy RSD
100 (mg/L, in plasma) (mg/L, in plasma) (%) (%, n=5)
labeled ISTD (3 mg/L CRP) Tryptic peptide from CRP 0.00805 0.00929 115 7.1
1.4 mg/L CRP in plasma in pooled human plasma 0.0161 0.0181 112 8.2
The challenge in peptide analysis and lipid profiling is to 50 RSD 0.69% 0.0322 0.0356 111 5.2
0.322 0.320 99 1.2
generate high data quality in complex samples. 3.22 3.01 93 0.7
0 32.2 29.8 93 1.0
For research use only. Not for use in diagnostic procedures. 2.50 2.75 3.00 3.25 min 322 337 105 0.9
Chromatogram of trypic digested peptide derived Quantitative results of AFVFPR (isotope labeled peptide)
from CRP using conventional flow rate spiked into trypic digested plasma

Lipid Mediator Profiling

Lipid mediators represent a class of bioactive lipids that are produced locally through 100 CV%
specific biosynthetic pathways in response to extracellular stimuli. Lipid mediators are
involved in many physiological processes, and their dysregulations have been often linked
to various diseases such as inflammation, infertility, atherosclerosis, ischemia, metabolic
syndrome, and cancer. To help further our understanding of lipid mediators in the disease
process we have created a method package designed to detect lipid mediators derived
from arachidonic acid cascade. 60

Using the lipid mediator method package the LCMS-8060 detected arachidonic acids Number of detected lipid mediators 80
metabolites in human serum over a wide dynamic range from sub nM to µM 40
concentrations. 5-HETE has shown the highest concentration which was 1 µM and the
lowest was 12-HHT (0.5 nM). In case of 8-iso-PGF 2α , its concentration was 0.1 nM.
20
5-HETE 12-HHT 8-iso-PGF2α
×10 7 ×10 3 ×10 4
319 > 115 279 > 179 353 > 193 10–20%
1.00
2.0 CV 8.0% (n=5) 0 0–10%
CV 0.9% (n=5) CV 5.4% (n=5) 1.5 LCMS-8050 LCMS-8060
0.75
1.5
1.0 Comparison of No. of detected lipid
0.50
1.0 mediators in human serum
0.5
0.25 0.5
0.00 0.0 0.0
17.0 18.0 min 15.0 min 9.0 10.0 min
MRM chromatograms of 5-HETE, 12-HHT and 8-iso-PGF2α

10 Liquid Chromatograph Mass Spectrometer LCMS-8060 11