Page 89 - Application Handbook - Liquid Chromatography

LAAN-A-LC-E275

Application Supercritical Fluid Extraction / Chromatography

News Application of Online SFE-SFC-PDA for Cleaning

Validation
No.L499

Cleaning validation is a process step that is extremely n Analytical Column Selection
important for ensuring high quality and safety at For SFC analysis, selection of the optimal column for the
pharmaceutical manufacturing sites. Cloth used for sample has a substantial effect on analysis reliability. We
surface wiping, called a swab, is used to wipe a given performed SFC separation of alkylbenzenesulfonate in four
part of a piece of manufacturing equipment, and different columns under the conditions shown in Table 1
analysis of the wiped area of the swab is performed by and Fig. 1, and chose the Shim-pack UCX-SIL analytical
using high-performance liquid chromatography (HPLC) column as it had the best peak shape. Based on an
or a total organic carbon analysis (TOC). Evaluations investigation of gradient profiles, we also found a relatively
using HPLC have been increasingly used in recent years steep gradient profile is suitable for quantitative analysis as
because HPLC enables determination of individual the properties of alkylbenzenesulfonate, which have
compounds. Prior to analysis, an extraction procedure different length of carbon chains, mean the significant
must be performed on the swab. Using supercritical peak broadiening if the gradient slope is not steep. Based
fluid extraction (SFE) as the pretreatment method allows on this information, we optimized analytical conditions
for simple and quick target component extraction. using the Shim-pack USX-SIL column and performed online
Using supercritical fluid chromatography (SFC) after SFE SFE-SFC analysis of a sample from a swab.
also means that analysis results can be obtained simply
by preparing the sample for SFE, which unifies the work
flow from pretreatment to analysis. Please see
Application News L496 for an overview of online SFE-
SFC. This article describes the process of column
selection using the Nexera-UC Chiral Screening System
as the first step in analysis of the target compound
alkylbenzenesulfonate.

Table 1 SFC Analytical Conditions for Column Selection
Column : Shim-pack UCX series columns (250 mm L. × 4.6 mm I.D., 5 μm)
(i) UCX-RP (ODS with polar group), (ii) UCX-GIS (ODS), (iii) UCX-SIL, (iv) UCX-DIOL
Mobile Phase : A: CO2; B: Methanol
Time Program : Shown in the figure
Flowrate : 3.0 mL/min
Column Temp. : 40 °C
Back Pressure : 15 MPa
Wavelength : 220 nm
Injection Vol. : Shown in figure

Injected sample:
1 µL of 1 % standard sample

Fig. 1 Comparison of SFC Separation of Standard Alkylbenzenesulfonate in Four Different Columns