Page 12 - LC-SFC_Pharma_Brochure

2-2. Sample Information
2-2. Sample Information
Three standards of chiral compounds (bromacil, α-methyl-α-acethyl-γ-butylrolactone, and methylclothiazide) were analyzed, as shown in Fig. 2.
These standards were prepared by dissolving in hexane and MTBE as needed at 1.0 mg/mL.

CI
H O O
CI N
O O
N
H2N
N O
+2
N S S
Br O − −
O O O
Bromacil α-Methyl-α-Acetyl-γ-Butyloractone Methylclothiazide
Fig. 2 Structural Formulas of Analyte Chiral Compounds

2-3. Chiral Columns
2-3. Chiral Columns
The iChiral-6 high-resolution polysaccharide-based columns (CHIRALPAK the iChiral-6 columns are compatible with the range of organic sol-
®
IA/IB/IC/ID/IE/IF) available from Daicel Corporation were utilized to vents specified here, these columns are applicable to method scouting
provide the high-resolution conditions for the displayed chiral com- analysis of chiral compounds.
pounds. Fig. 3 shows the respective column functional groups. Since

OR OR
(A) CHIRALPAK IA (150 mmL. × 4.6 mmI.D., 5 µm) O O O
®
Amylose tris(3,5-dimethylphenylcarbamate) OR OR O n OR O n
Immobilized to Immobilized to
(B) CHIRALPAK IB (150 mmL. × 4.6 mmI.D., 5 µm) silica gel silica gel
®
Cellulose tris(3,5-dimethylphenylcarbamate)
Amylose Derivatives Cellulose Derivatives
®
(C) CHIRALPAK IC (150 mmL. × 4.6 mmI.D., 5 µm)
Cellulose tris(3,5-dichlorophenylcarbamate) R : H H
N CH 3 H N CH 3
N
®
(D) CHIRALPAK ID (150 mmL. × 4.6 mmI.D., 5 µm) O O O
Amylose tris(3-chlorophenylcarbamate) CH 3 Cl CH 3
(A) CHIRALPAK ® IA (D) CHIRALPAK ® ID (B) CHIRALPAK ® IB
®
(E) CHIRALPAK IE (150 mmL. × 4.6 mmI.D., 5 µm)
H H H N Cl
Amylose tris(3,5-dichlorophenylcarbamate) N Cl N
O O O
CH 3
(F) CHIRALPAK IF (150 mmL. × 4.6 mmI.D., 5 µm) Cl Cl Cl
®
Amylose tris(3-chloro-4-methylphenylcarbamate) (E) CHIRALPAK ® IE (F) CHIRALPAK ® IF (C) CHIRALPAK ® IC
Fig. 3 Chiral Selector
2-4. Separation Conditions
2-4. Separation Conditions
For the mobile phases, eight different solvent mixtures were used, combinations and 6 different columns, the 48 unique separation
consisting of various combinations of hexane, 2-propanol, ethanol, conditions yielded an exhaustive search for the conditions suitable
dichloromethane, and methyl-t-butyl ether. Details of the separation for the separation of each of the chiral compounds.
conditions are shown in Table 1. Using a total of 8 mobile phase
Table 1 Analytical Conditions
Separation Conditions Mobile Phase Flow Rate Analysis Time Other
No.
1 Hexane / 2-Propanol = 9 / 1(v/v) 3 mL/min 9 min
2 Hexane / 2-Propanol = 6 / 4(v/v) 3 mL/min 9 min
3 Hexane / Ethanol = 8 / 2 (v/v) 3 mL/min 14 min
4 Ethanol 1 mL/min 18 min Column temperature: 40°C
Injection volume: 10 µL
5 Hexane / Dichloromethane = 9 / 1(v/v) 3 mL/min 4 min Detection: 230 nm
6 Dichloromethane / Ethanol = 100 / 2(v/v) 3 mL/min 4 min
7 Hexane / Methyl tert-Butyl Ether = 9 / 1(v/v) 3 mL/min 4 min
8 Methyl tert-Butyl Ether / Ethanol = 9 / 1(v/v) 3 mL/min 4 min
Note: For analysis of methylclothiozide, 0.1% diethylamine was added to each mobile phase.

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