Page 11 - LifeScience Solution for Lipid and Lipidome

Analysis of Glycerophospholipids

Data Subcutaneous Tissue Mass Imaging of
Phospholipidosis is an excessive accumulation of phospholipids in the lysosomes
of cells that is induced by pathological conditions or medications. It is
In the first dimension, phosphatidylserine (PS) was separated from other phospholipids by normal-phase separation. After dilution characterized by lamellar bodies observed inside lysosomes. Many drugs with a
and trapping, components were further separated by reverse-phase separation based on acyl group chain length and then cationic amphiphilic structure (cationic amphiphilic drugs) are known to cause Analysis of
detected using LC-MS. This allowed analysis of the molecular species of phosphatidylserine (PS) while inhibiting effects from other drug-induced phospholipidosis. The clinical problem with drug-induced Comprehensive
phospholipids (Fig. 1). phospholipidosis is that the drugs that induce the condition can cause adverse Glycerophospholipids
side-effects in humans, such as disorder of the liver, lungs, spleen, or other
organs. Consequently, drug-induced phospholipidosis is attracting considerable
(×100,000) attention with respect to drug safety. From a clinical perspective, it is important
10.0
PC 1 D chromatogram to evaluate the organs for any functional disabilities and search for biomarkers.
st
8.0 However, there is still much that is unknown about the induction mechanism of
6.0 drug-induced phospholipidosis. From a basic research perspective, the major Technology
4.0 PG PE issue is determining the induction mechanism. Supercritical Fluid Lipid Analysis Using
2.0 PI
PS
0
0 5 10 15 20 25 30 35 min
(×1,000) (×10,000)
6.0
16.0 1 D chromatogram 1 D chromatogram 2 D chromatogram of Co-Sense Technology Analysis of
st st
nd
12.0 4.0 PS fraction
8.0 Co-Sense series systems only inject a portion of the solution Pump unit for first Glycerophospholipids
dimension separation
4.0 2.0 separated by the first-dimensional column into the First dimension
Autosampler
0 0 second-dimensional column. Compared to comprehensive separation column
5 10 15 20 25 30 min 0 5 10 15 20 25 30 min 2D-LC systems, this allows use of a longer second-dimension
column that is able to separate components more efficiently.
Fig. 1 Analysis of Phosphatidylserine (PS) by Co-Sense for Impurities Concentration column
Therefore, it is especially useful for analyzing the molecular
species of target phospholipids in more detail, such as Pump unit for on-line Blood Serum
phosphatidylserine (PS). concentration Mediators in Human Analysis of Lipid
Results from the quantitative analysis of phosphatidylserine (PS) molecular species obtained using the LCMS-8050 are shown in Table The Co-Sense for Impurities system can be used to separate
1. If only reverse-phase separation is used, ion suppression and enhancement are found (shown in red in the table), but using the phospholipids by normal-phase LC in the first dimension and Second dimension
Co-Sense for Impurities system allows elimination of coexisting substances and achieves more accurate quantitative results. further separate them according to the carbon chain length of Detector separation column
fatty acids by reverse-phase LC in the second dimension. This Pump unit for second
dimension separation
makes it possible to fully automate the previously manual
process of TLC separation, recovery from TLC plates, and then
Table 1 Quantitative Analysis Results for Phosphatidylserine (PS) Molecular Species
separation by reverse-phase LC. of Glycolipids
Area Ratio (%) Structural Analysis
# Compounds Conventional RP LC 2D -LC
Sample 1 Sample 2 Sample 3 Sample 1 Sample 2 Sample 3
1 16:0-16:0 PS 93.82 87.94 66.11 96.36 99.88 100.02
2 18:0-18:0 PS 82.99 139.4 113.7 104.89 97.69 91.26
3 16:0-20:4 PS 101.17 81.72 69.00 115.78 101.28 89.71
4 18:0-20:4 PS 95.07 110.4 83.55 112.45 104.91 91.67
5 18:0-22:6 PS 114.68 117.3 54.75 103.30 116.60 101.30 Analysis of Fatty Acid
6 18:1-18:1 PS 100.77 139.4 105.4 96.83 84.69 96.30 Content of Human ES Cells Composition in Overall Lipid
7 16:0-16:0 PG 99.20 89.06 70.86 89.02 77.90 82.29
8 16:0-18:1 PG 106.30 90.63 88.89 103.40 89.22 86.91
9 16:0-20:4 PG 106.60 93.12 108.31 92.95 83.80 95.06
10 16:0 -18:2 PG 111.36 93.16 100.80 110.94 93.70 100.13
11 18:0 -18:0 PG 109.47 91.78 80.70 92.50 83.22 83.19
12 18:0 -20:4 PG 107.14 91.22 79.98 112.72 98.66 83.67
13 18:0 -22:6 PG 95.24 85.02 77.39 86.13 77.35 84.73
14 22:6 -22:6 PG 112.99 94.39 102.40 94.40 80.37 84.88 High-Performance Liquid Chromatograph High-Throughput Liquid Chromatograph Mass Spectrometer Esters by GC Acid Methyl
Co-Sense for Impurities LCMS-8060 Analysis of Fatty

Reference: Co-Sense Series brochure (C196-E089), LCMS-8060 brochure (C146-E286)

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