Normal Phase HPLC Column COSMOSIL SL-II

Transcription

1 Normal Phase HPLC Column COSMOSIL SL-II High purity silica gel (>99.99%) with special treatment Suitable for preparative separation Ultra-pure silica gel of more than 99.99% purity is used for the COSMOSIL SL-II packed column series. This column provides improved separation and reproducibility for compounds with carbonyl or phenol hydroxyl groups, which are often problematic to separate using conventional silica gel columns because of interference of metallic impurities. The followings are applications of COSMOSIL SL-II column on organic acids, acid amides and phenols. COSMOSIL SL-II provides improved separation for these compounds without ionic additives by using mobile phases of hexane and ethanol. Comparison with Competitors Columns in Separation of Organic Acids and Acid Amides Flow Rate 4.6 mm I.D. x 50 mm Ethanol Hexane = ml/min 30 UV 54 nm 0.3 AUFS. Benzoic Acid (4.0 μg). Salicylic Acid (6.0 μg) 3. Salicylamide (4.0 μg) CO CO CONH. Benzoic Acid. Salicylic Acid 3. Salicylamide Company A Company B Company C Company D Company E Company F

2 Comparison with Competitors Columns in Separation of Phenols Flow Rate 4.6 mm I.D. x 50 mm Ethyl Acetate : Hexane = ml/min 30 o C UV 54 nm 0.6 AUFS. p-xylene. Chrysazin 3. Paeonol 4. p-cresol (0.004 μ ) (0. μg) (0.4 μg) (0.004 μ ) CH 3 O COCH 3 HO CH 3 O OCH 3. p-xylene. Chrysazin 3. Paeonol CH 3 4. p-cresol Company A Company B Company C Company D Company E Company F Comparison of Purity of Silica Gel with Competitors columns Column Purity of Silica Gel Particle Size COSMOSIL SL-II 99.99% and more 5 m Company A No guaranteed value (metallic impurities 5 ppm or less) 5 m Company B No guaranteed value (high purity) 5 m Company C % 5 m Company D % 5 m Company E No guaranteed value 5 m Company F No guaranteed value 5 m

3 Scaling up from Analytical to Preparative Separation Normal phase chromatography, with non-polar mobile phase and low boiling point is used for preparative separation because solvent removal is generally easier for normal phase chromatography than for reversed chromatography. The followings are how to scale up from analytical (4.6 mm I.D.) to preparative (0.0 mm I.D.) separation. Step Step Step 3 Mobile phase is optimized by using COSMOSIL SL-II (4.6 mm I.D. x 50 mm) As mobile phase, highly soluble solvent for sample is suitable. Maximum load capacity is optimized by using COSMOSIL SL-II (4.6 mm I.D. x 50 mm) Maximum injection volume 56 µl (8 mg) Preparative separation by using COSMOSIL SL-II (4.6 mm I.D. x 50 mm) Maximum injection volume,050 µl (55 mg) (9 times higher than 4.6 mm I.D.) Flow Rate Ethyl Acetate : Hexane = 0 : mm I.D..0 ml/min 0.0 mm I.D. 8.9 ml/min Room UV 54 nm. -Naphthol. -Naphthol Preparative Scale Comparison of Normal Phase and Reversed Phase Normal phase chromatography is more suitable for preparative separation than reversed phase chromatography as larger amount of sample can be loaded in normal phase chromatography. The following are preparative scale comparison of normal phase (COSMOSIL 5C8-MS-II) and reversed phase (). COSMOSIL 5C8-MS-II 0.0 mm I.D. x 50 mm Methanol : Water = 50 : mm I.D. x 50 mm Mobile phase Ethyl acetate : Hexane = 0 : 90 Maximum Load Capacity : 58 mg 55 mg Maximum Load Capacity:

4 Preparative HPLC as an Alternative Approach to Open Column Chromatography Open column chromatography has limitations, such as low separations and long separation time. To overcome these disadvantages, preparative HPLC chromatography can be an alternative approach. Preparative HPLC offers high separations and shorter analysis time. Furthermore it is environmentally friendly as it can be used repeatedly and there is no need to dispose silica gel. Open Column Chromatography Preparative HPLC Flow Rete Silica Gel 60 (50 35 mesh) 0.0 mm I.D. x 300 mm Ethanol : Hexane = 50 : 50.0 ml/min Room ( ) UV 54 nm 0.6 AUFS. p-xylene. Cinnamyl Alcohol 3. p-nitrobenzyl Alcohol (0.4 mg) (0.005 mg) (0.03 mg) Flow Rete 0.0 mm I.D. x 50 mm Ethanol : Hexane = 5 : 95 8 ml/min 30 UV 54 nm 0.6 AUFS. p-xylene (8.6 mg). Cinnamyl Alcohol (0.4 mg) 3. p-nitrobenzyl Alcohol (.0 mg) * Analyzed by closed column at the same condition as open chromatography. * Length of column = Bed height of open column chromatography. Open Column Chromatography Preparative HPLC Separation Capacity (Theoretical Plate Number) ) approx fold approx. 0,000 Preparative Time approx.0 min. /8 6 min The number of use /8 approx.,000 Loading Capacity ) 0.03 mg 70-fold.0 mg Repeatability Quantitative Performance Difficult Easy Adjustment of Column Necessary Unnecessary Disposal of Silica Gel Necessary Unnecessary Pretreatment of Unnecessary Necessary Equipment Unnecessary Necessary Note: ) Compared by peak of p-nitrobenzyl Alcohol. Ordering Information Particle Size: 5 m (Analytical/Preparative Columns) Product Name Product Number Packed Column Gurad Column 4.6 mm I.D. x 50 mm mm I.D. x 00 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 0 mm mm I.D. x 0 mm mm I.D. x 0 mm mm I.D. x 50 mm mm I.D. x 50 mm Particle Size: 5 m (Preparative Columns) Product Name Product Number COSMOSIL 5SL-II Packed Column COSMOSIL 5SL-II Gurad Column 8.0 mm I.D. x 50 mm mm I.D. x 50 mm mm I.D. x 500 mm mm I.D. x 50 mm mm I.D. x 50 mm Particle Size: 3 m (Fast LC Columns) Product Name Product Number COSMOSIL 3SL-II Packed Column Other size may be available. Please enquire. 4.6 mm I.D. x 0 mm mm I.D. x 50 mm mm I.D. x 00 mm NACALAI TESQUE, INC. Nijo Karasuma, Nakagyo-ku, Kyoto , JAPAN TEL : +8-(0) FAX : +8-(0) Website : info.intl@nacalai.com For research use only, not intended for diagnostic or drug use. 0

5 I. HPLC Columns COSMOSIL SL-II II. UHPLC Columns SL-II III. Preparative Packing Materials COSMOSIL SL- IV. Related Products Comparison with Old Type Column: Column size: 4.6mmI.D.-50mm Mobile phase: Ethanol/Hexane = 0/90 Flow rate:.0 ml/min : 30 C : UV54nm 5SL-II 5SL (old type) : ; Benzoic Acid (4.0 g) ; Salicylic Acid (6.0 g) 3; Salicylamide (4.0 g) V. Applications NACALAI TESQUE, INC VI. Technical Notes VII. Index COSMOSIL SL- - Comparison with Medium-Pressure Columns Column: Column size: Mobile phase: Ethanol/Hexane = 5/95 Flow rate: 0 ml/min : room temperature : UV54nm : ; p-xylene (8.3mg) ; Cinnamyl Alcohol (.7mg) 3; p-nitrobenzyl Alcohol (4.mg) 5SL- 0mmI.D. x 50mm Pressure:.3MPa I company cartridge 30 m silica gel 6mmI.D. x 04mm Pressure: 0.5MPa I company cartridge 60 m silica gel 6mmI.D. x 04mm Pressure: 0.5MPa NACALAI TESQUE, INC 34

6 - - Ordering Information COSMOSIL 3SL I. HPLC Columns II. UHPLC Columns III. Preparative IV. Related Products V. Applications VI. Technical Notes VII. Index Packing Materials