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(Reference) Inuence of organic solvent composition in mobile phase on the retention time
Selectivity of COSMOSIL Packing Materials
1
3
5 Over 7,000 applications COSMOSIL Applications 69:31
0 5 10
8.44min
Average
15
Easy to search 0.66min
approx. 120 approx. 300
Pore Size
68:32 9.10min
Specific
approx. 300 m2/g approx.150m2/g 0 5 10
SurfaceArea COSMOSIL Application Data 5C18-MS-II
2.5C18-MS-II 0.61min
(4.6mmI.D.-150mm)
Column: COSMOSIL C18-MS-II (3.0mmI.D.-75mm)
Pressure: 3.7MPa
Column size:
Flow rate:
3.0mmI.D.-75mm
1.0 ml/min
Pressure: 10.9MPa
10.9MP
67:33 9.71min
Mobile phase: Acetonitrile/ H2O = 70/30 0 5 10
H3C Pressure: 10.9MPa
Temperature:
Detection:
40C
UV254nm
0.73min
H3C Sample:
Sample 1; Benzene (1.67mg/ml)
Bonded NO2
2; Toluene (1.67mg/ml) 66:34 10.44min
Phase
3; Ethylbenzene
4; n-Propylbenzene
(1.67mg/ml)
(1.67mg/ml)
0 5 10
O 5; n-Butylbenzene (1.67mg/ml)
6; Amylbenzene (1.67mg/ml)
Injection Vol. 1.0l
H3C
Si
CH3
Si Si
OH
Si Si Si
H3C
Si
CH3 H3C
Si Si Si (CH2)n-H
(C
NACALAI TESQUE, INC
Special attention should be paid to measure correct amount Column 5C18-MS-II 4.6 mmI.D. x 150 mm
OH H3C CH3 H3C CH3 H3C CH3 CH3 H3C CH3 OH
- n=0-5
of organic solvent as retention time is significantly changed Flow rate 1.0ml/min
Octadecyl Octadecyl Dococyl Octyl Butyl Cholesteryl Naphtylethyl Pyrenylethyl Nitrophenylethyl Secondary Octadecyl
Group Group Group Group Group Group Group Group Group Triazole /Tertiary Amine Group by 1% different composition. Detection 254nm 0.16AUFS
Bonding Temperature 30oC
Monomeric Polymeric Monomeric - Polymeric Sample 1. Uracil
Type
Hydrophobic 2. Naphthalene
Interaction
Hydrophobic
Hydrophobic - Interaction
Interaction Hydrophobic Interaction Hydrophilic
Main Interaction - Interaction Hydrophobic
Hydrophobic Interaction
Interaction Molecular
Shape -
Interaction
Dispersion
Force
Interaction Anion
Exchange
Interaction 2) Organic Solvent / Buffer Mixed Mobile Phase
Selectivity Dipole-dipole
Charge interaction
transfer (e.g.1) Preparation of 20 mmol/l phosphate buffer (pH2.5)
Interaction
End-capping
Near-perfect Treatment
Treatment
Column: COSMOSIL C18-MS-II Prepare 20 mmol/l sodium dihydrogenphosphate
Carbon
Load
approx. 14.5% approx. 16% approx. 11% approx. 11% approx. 19% approx. 10% approx. 7% approx. 20% approx. 11% approx. 18% approx. 9% -
Column size: 3.0mmI.D.-75mm aqueous solution
For hydrophilic Distilled water
Excellent Multi-
Strong acid
resistance
compounds High Stronger Strongest Dipole-
For highly
polar analysis For accharide
analysis
High
recovery rate
Flow rate: 1.0 ml/min
Features for Basic purpose C18 Compatible molecular - - dipole
compounds Column For acid
compounds
with 100%
water based
shape
selectivity interaction interaction interaction
Unique anion-
exchange
High
durability
Acid-
resistant Mobile phase: Acetonitrile/ H2O = 70/30
mechanism
mobile phase
Pressure: 10.9MPa
I.D.
5. When mix with organic solvent, mix by volume ratio.
Composition table for mobile phase 1L Composition table for mobile phase 1L Attention; The solid may precipitate after mixing.
Inner Diameter of Column (scale down and up) (Methanol : water) (Acetonitrile : water) Phosphate Buffer Solution (pH 7.0) (5x) (Product No, 08968-81) (Ready to use ) is avaialble from Nacalai Tesque.
Methanol / Water Methanol (g) Distilled Water (g) Acetonitrile / Water Acetonitrile (g) Distilled Water (g)
Inner Diameter 90 : 10 (v/v) 711.9 99.8 90 : 10 (v/v) 707.4 99.8
1.0 2.0 3.0 4.6 10 20 28 50 (e.g.3) Preparation of 5 mmol/l Sodium 1-hexanesulfonate, 20 mmol/l phosphate buffer (pH2.5)
(mm I.D.) 80 : 20 (v/v) 632.8 199.6 80 : 20 (v/v) 628.8 199.6
Flow Rate (ml/min) 0.05 0.2 0.4 1 5 19 37 70 70 : 30 (v/v) 553.7 299.5 1. Prepare 5 mmol/l Sodium 1-hexanesulfonate, 20 mmol/l phosphate buffer (pH2.5) aqueous solution (Dissolve 10
70 : 30 (v/v) 550.2 299.5
Detector Cell 60 : 40 (v/v) 474.6 399.3 ml of Sodium 1-hexanesulfonate (0.5 M solution) (Product No. 31532-06) and 2.40 g of sodium
for Semi-micro for Analytical for Preparative 60 : 40 (v/v) 471.6 399.3
Injector dihydrogenphosphate, Anhydrous (Product No. 31720-65) in distilled water to make 1L solution.).
50 : 50 (v/v) 395.5 499.1 50 : 50 (v/v) 393 499.1
Inner Diameter of 2. Prepare 5 mmol/l Sodium 1-hexanesulfonate, 20 mmol/l phosphate aqueous solution (Dissolve 10 ml of sodium
0.05 0.1 0.2-0.3 1 40 : 60 (v/v) 316.4 598.9 40 : 60 (v/v) 314.4 598.9
Pipe (mm) 1-hexanesulfonate (0.5 M solution) (Product No. 31532-06) 2.31g of phosphoric acid (Purity: 85%), (Product No.
30 : 70 (v/v) 237.3 698.7 30 : 70 (v/v) 235.8 698.7
Solvent 08964-92) in distilled water to make 1L solution.).
Preparative Preparative
Application LC-MS solvent saving
saving with
Standard
Preparative
(medium
Preparative
(Super large 20 : 80 (v/v) 158.2 798.6 20 : 80 (v/v) 157.2 798.6
standard (small scale) (large scale) 3. Adjust the pH to 2.5 by mixing 1 with 2.
scale) scale) 10 : 90 (v/v) 79.1 898.4
system 10 : 90 (v/v) 78.6 898.4 4. Filter under reduced pressure to remove insoluble substance (0.45 m or smaller pore size is recommended.).
Surface Ratio with Caution : Methanol and acetonitrile are hazardous substances. Always process in a laboratory hood and wear an eye protection and a mask. Attention; Filter solids from the solution to prevent clogging to pump and columns.
0.05 0.19 0.43 1.00 4.73 18.90 37.05 118.15
4.6 mm I.D.
5. When mix with organic solvent, mix by volume ratio.
Particle Size (m) 2.5, 3 or 5 3 or 5 5 15 or more
Attention; The solid may precipitate after mixing.