Page 52 - LC-SFC_Pharma_Brochure

(×100,000)
3.0 Reserpine
10 µg/L
5 µg/L
0.5 µg/L ŶPeaks
2.0 1. Phosphatidyl Choline
1 2. Lyso Phosphatidyl Choline
2 3. Phosphatidyl Ethanolamine
1.0 4. Lyso Phosphatidyl Ethanolamine
No splitting 3 5. Phosphatidyl Glycerol
4 6. Phosphatidyl Inositol
Flow path split 5
0.0
(Split ratio, Vent:MS = 4:1)
6
0. 0 0 .5 1. 0 1 .5 min
0 2. 5 5 .0 7.5 10.0 1 2.5 min
Column : Shim-pack UC-Diol 4.6 mm I.D. × 150 mm L.
Modiÿer : MeOH Column : Shim-pack UC-Diol 4.6 mm I.D. × 150 mm L. 5 µm
Modiÿer conc. : 30 % Modiÿer : MeOH with 0.1 % w/v ammonium formate
Flow rate : 3.0 mL/min Modiÿer conc. : 10 % (0 min) ĺ 40 % (10–15 min)
Temperature : 40°C Flow rate : 3.0 mL/min
Back pressure : 10 MPa Temperature : 40°C
Injection volume : 1 µL Make-up solution : MeOH with 0.1 % w/v ammonium formate
Sample : Reserpine (0.5, 5, 10 °g/L) Make-up °ow rate : 0.1 mL/min
Detection : TQ mass spectrometer ESI(+) m/z 609.3 > 195.0 Back pressure : 10 MPa
Detection : TQ mass spectrometer (ESI)
Fig. 8 Sensitivity With and Without Flow Path Splitting
Fig. 9 Phospholipid Analysis
Table 2 MS Reproducibility With and Without Flow Path Splitting
Fig. 10 shows an example analysis of pesticides of a wide range of po-
Injection Retention time Area Height
volume larities—from hydrophobic to hydrophilic—using the Shim-pack
(µL) Ave. %RSD Ave. %RSD Ave. %RSD UCX-RP column. The Shim-pack UCX-RP column is unique in having
0.1 0.359 0.64 6,583 18.83 2,361 17.29
a stationary phase that combines octadecyl and polar functional
Flow path split 1 0.356 0.25 81,467 4.26 26,656 3.88 groups. This stationary phase is able to retain a wide range of com-
2 0.355 0.32 156,831 2.18 49,721 3.28 pounds, including both hydrophobic and hydrophilic compounds.
0.1 0.356 0.09 16,264 6.18 7,673 6.17 This column allows the simultaneous analysis of pesticides that were
No splitting 1 0.353 0.05 155,170 2.43 71,971 2.23 previously difÿcult to analyze without changing the analytical condi-
2 0.35 0.07 325,739 1.16 142,350 1.19 tions, thereby providing improved analytical efÿciency.
Column : Shim-pack UC-Diol 4.6 mm I.D. × 150 mm L. 5 µm
Modiÿer : MeOH with 0.1 % w/v ammonium formate 1
Modiÿer conc. : 30 % 3 2 4
Flow rate : 2.0 mL/min
Temperature : 40°C 5
Back pressure : 10 MPa Hydrophobic 6
Injection volume : 1 µL 7
Detection : TQ mass spectrometer ESI(−) m/z 351.20 > 271.20 (prostaglandin 100 µg/L) 8
9 10
4. Shim-pack UCX Series Columns for SFC 11
12
14
Because of the high diffusivity of the mobile phase used in SFC, the Hydrophilic 13
separation behavior substantially changes based on the column sta-
tionary phase and modiÿers used. The Shim-pack UCX series columns
are designed for SFC and encompass eight different stationary 0. 0 5 .0 10.0 min
phases, as shown in Table 3. This allows the columns to accommo-
date the separation of a wide variety of compounds. No. Compound log P
1 Carbofuran 7.4
Table 3 Shim-pack UCX Series Columns 2 Ethofenprox 6.9
Functional group Pore Surface Carbon 3 Fenpropathrin 6.0
size area content
Shim-pack UC-RP Octadecyl group + polar functional group — 9% 4 Pyriproxyfen 5.0
Shim-pack UC-GIS II Octadecyl group 11% 5 Pyraclostrobin 4.0
Shim-pack UC-Diol Diol group 20% 6 Linuron 3.0
Shim-pack UC-Sil — — 7 Aminocarb 1.9
10 nm
Shim-pack UC-Amide Carbamoyl group 450 m 2 /g 18% 8 Ethoxysulfuron 1.0
Shim-pack UC-NH2 Aminopropyl group 8% 9 Halosulfuron methyl 0.0
Shim-pack UC-Phenyl Phenethyl group 9.5% 10 Bentazone −0.5
Shim-pack UC-CN Cyanopropyl group 14% 11 Chlorsulfuron −1.0
12 Rimsulfuron −1.5
Fig. 9 shows an example analysis of phospholipids using the Shim-pack −1.8
UCX-Diol column. This column allows separation of phospholipids by 13 Nicosulfuron
class, as with normal phase LC. Phospholipids can also be separated 14 Vamidothion −4.2
by molecular species using the same modiÿer conditions paired with Column : Shim-pack UC-RP 4.6 mm I.D. × 150 mm L. 5 µm
: MeOH with 0.1 % w/v ammonium formate
Modiÿer
a different column, such as the Shim-pack UCX-GIS II, which has an Modiÿer conc. : 0 % (0 min) ĺ 10 % (11 min) ĺ 30 % (14 min) ĺ 40 % (14.01–17 min)
octadecyl group stationary phase. Using different stationary phases Flow rate : 3.0 mL/min
: 40°C
Temperature
but the same mobile phase, SFC can be used to recreate the retention Make-up solution : MeOH with 0.1 % w/v ammonium formate
behaviors observed with normal phase and reverse phase HPLC, pro- Make-up °ow rate : 0.1 mL/min
: 15 MPa
Back pressure
viding a variety of other separation behaviors. This is of substantial Detection : TQ mass spectrometer (ESI)
beneÿt for the analysis of complex samples. Fig. 10 Pesticide Analysis
3