Page 90 - Application Handbook - Liquid Chromatography

Application No.L499
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n Online SFE-SFC Analysis of a Swab Containing
Alkylbenzenesulfonate uV (× 1,000,000)
We investigated column selection by the scouting 3.0
system, chose the Shim-pack UCX-SIL analytical column, 100 ng of alkylbenzenesulfonate
optimized each analytical condition for online SFE-SFC 2.5
analysis, then performed analysis using the conditions 2.0
shown in Table 2 below.
1.5 1st
Table 2 Analytical Conditions for Online SFE-SFC
1.0 2nd
[Sample Preparation] 0.5 3rd
A total of 10 to 500 μg standard samples in methanol were dropped 4th
onto swabs.
The swabs were enclosed into an extraction vessel and set to the SFE unit. 0.0
[Static Extraction] 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 min
Extraction Time : 3 min
Mobile Phase : A: CO2; B: 0.1 % (w/v) Ammonium Formate in Methanol
B Conc. : 10 % Fig. 3 Confirmation of Online SFE Extraction Efficiency
Flowrate : 3.0 mL/min
Back Pressure : 15 MPa
[Dynamic Extraction]
Extraction Time : 3 min
Mobile Phase : A: CO2; B: Methanol mAU
B Conc. : 10 % Static Dynamic SFC
Flowrate : 3.0 mL/min 3.0 Extraction Extraction Analysis
Back Pressure : 15 MPa
[SFC] 2.5
Column : Shim-pack UCX-SIL (250 mm L. × 4.6 mm I.D., 5 μm) Alkylbenzenesulfonate
Mobile Phase : A: CO2; B: Methanol 2.0
Time Program : 10 %B (0-2 min), 10-60 %B (2-7 min),
60 %B (7-9 min), 10 %B (9-13 min) 1.5
Flowrate : 3.0 mL/min 500 µg
Column Temp. : 40 °C 200 µg
Back Pressure : 15 MPa 1.0
Wavelength : 220 nm 100 µg
0.5 50 µg
The peak for the surfactant alkylbenzenesulfonate was 0 20 µg
well-separated and detected as shown in Fig. 2 below. 10 µg
Fig. 3 shows the results of performing repeated SFE-SFC -5.0 -2.5 0.0 2.5 5.0 7.5 min
analyses from the same swab to which had been added
an equivalent of 100 ng of alkylbenzenesulfonate. Since
there was almost no alkylbenzenesulfonate peak evident
from the second and later sample extractions, the
extraction procedure was almost entirely complete after
the first SFE. Fig. 4 shows the results of adding amounts
of alkylbenzenesulfonate to swabs in the range of 10 to
500 μg, and checking linearity. Within this range, the
coefficient of determination that represents linearity was
0.996. Fig. 5 shows the result of five consecutive
analyses of separate swabs to which were added 100 μg
of alkylbenzenesulfonate. Considering the process
including extraction, the repeatability of retention times Fig. 4 Linearity of Online SFE-SFC Analysis Using a Swab
was 0.19 %RSD, and repeatability of peak area was
5.76 %RSD. Based on these results, we confirmed the
usefulness of the Nexera-US Online SFE-SFC System in mAU
SFC
Dynamic
Static
this application. 2.0 Extraction Extraction Analysis
Alkylbenzenesulfonate
1.5
200 µg of alkylbenzenesulfonate added in swab
1.0
Swab blank 0.5

0
-5.5 -2.5 0.0 2.5 5.0 7.5 min

Fig. 5 Repeatability of Online SFE-SFC Analysis Using a Swab
-5.0 -2.5 0 2.5 5.0 7.5 min
Fig. 2 Online SFE-SFC Analysis of Alkylbenzenesulfonate Note: Swab samples were provided by Eisai Co., Ltd.
First Edition: Jan. 2016

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