[ PROCESS PURIFICATION ] ™™ DAISOGELDAISOGELPacking Materials for Process Purification DAISOGELPacking Material for Process Purification - offered by Waters Waters Corporation and DAIso CO., LTD., Osaka, Japan, have jointly entered into a collaboration to globally market and supply DAIsoGEL™ bulk packing materials for process chromatography. By combining the purification expertise of both organizations, pharmaceutical and biopharmaceutical companies’ scaling processes for lab-scale to pilot plant or process-scale manufacturing, can now benefit from an unparalleled level of technical and product support. Daisogel Packing Materials . Silica-based packing materials for pharmaceutical and biopharmaceutical chromatographic purifications . High mass loading and excellent resolution . Excellent durability with extended acidic and alkaline resistance . Available in multiple kilograms Daisogel Analytical and Preparative HPLC Columns Packed by Waters . Analytical columns for screening and method development of isolation and purification methods . Preparative columns packed with Waters patented Prep OBD™ Technology for scaling up [3] DAISOGEL Columns Packed with Waters OBD Technology ColUMN PERFORMANCE – IdENTIFYING THE PRoblEM Laboratory-scale HPLC purification presents many challenges to the chromatographer. Inconsistency in column-to-column performance and OBD Optimized Bed Density lifetimes often result in lost samples, repeat purification runs and poor scalability from small to larger volume columns. Common HPLC prep High-pressure Slurry Packing Axial-ram Slurry Packing column failures are due to mechanical instabilities, especially voids in the Localized Bed Expansion Near Inlet Localized Compression Near Inlet packed bed. The patented OBD Technology1 combines optimized packing processes with a unique set of column designs ensuring increased lifetime OBD Process and better reproducibility. Optimum Bed Density Throughout When a column is used in a preparative HPLC system, it must remain stable during operation. The bed must be packed sufficiently dense to withstand the compressive fluid forces encountered during use. In the case of analytical column dimensions, the necessary packed bed density can be generally achieved using traditional slurry packing methods. As the diameter of the column increases, it becomes increasingly difficult to reach the bed density required for stable, long-term performance, especially when using small particles. The optimum bed density is Optimized Bed Density is achieved by applying pre-determined dependent upon the specific properties of the chromatographic particles axial force only at the column inlet. and column dimensions. 1US Patent # 7,399,410; UK Patent # GB2408469 OBD ColUMN DESIGn – IMPROVED MEchANICAL STABILITY Waters combines high-pressure slurry packing with a carefully calculated axial compression element localized at the less-dense inlet end of the bed. With careful tuning of the packing process for each particle type and column geometry, the Prep OBD design and process results in predictable, uniform density profiles throughout the column. Established procedures ensure that the column bed is not over-compressed or disrupted during the final column packing process. Excess axial compression applied at the inlet can break particles, build bridges, and lower local bed permeability. Exploded Prep OBD Column Hardware The patented OBD column hardware design incorporates chemically inert seals made to prevent leaks and a pair of specially designed filter housings with frits and distributors built to withstand high pressures. [4] Daiso CO., Ltd. Daiso CO., Ltd., headquartered in Osaka, Japan, is a technology-oriented chemical DAIsoGEL PACKING MATERIAL company supporting industries including medicine, agriculture, IT, machinery, Daiso CO., Ltd. is one of the leading silica gel manufacturers for liquid chromatography, using inorganic synthesis technology. Since textiles, and construction through 1992, Daiso has added large-scale manufacturing capability for innovative products. Daisogel media to fully support the process purification laboratories worldwide. Daisogel phases are used for various applications including medicines and pharmaceuticals, agricultural chemicals, PRodUCT OFFERING INclUDES: food ingredients, biomolecules, liquid crystals and functional dyes. DAISOGEL packing materials Daisogel packing materials have been designed to meet the needs of process purification. These speherical silica-based packing materials . Pharmaceutical intermediates allow for high mass loading and maximum resolution which can result . Fine chemicals in reducing the number of repeated injections. Through high density bonding and endcapping, Daisogel packing materials have shown . Proprietary functional chemicals extended life and durability. These benefits have helped customers in reducing their overall purification costs. Soda products and Chlor-Alkali . Synthetic and fabricated resin products QUALITY Daiso quality control system is based on ISO 9001 for the silica gel manufacturing plant and has a DMF (Drug Master File) on file www.daiso-co.com with the FDA (Food and Drug Administration) for the Daisogel packing materials. Starting with the raw materials, the quality of Daisogel packing material is strictly controlled at each manufacturing step. Each Daisogel batch is analyzed for pore size, surface area, pore volume and particle size, along with measurements of the silica gel pH, volatile matter, and metal impurities. Daisogel packing materials are designed to perform with consistent selectivity from lot-to-lot and between particle sizes. [5] DAISOGEL DAISOGEL SP-120-ODS-BP . Silica-based 120Å C18 phase Packing Materials . Suitable for hydrophilic compounds separation . Longer lifetime in aqueous eluents . Different selectivity from ODS-RPS DAISOGEL SP-120-ODS-RPS ODS-BP phases are designed to show extended selectivity for . Silica-based 120Å C phase 18 hydrophilic and polar compounds which are either poorly or not . High coverage and exhaustive endcapping retained on other phases. Typical applications are separations of biomolecules and metabolites such as oligosaccharides, amino ODS-RPS packings featuring maximum surface coverage are the ideal choice for a acids, small peptides, nucleotides and organic acids. They are wide variety of organic compounds. Carefully controlled full endcapping leads to endcapped and show similar selectivity as conventional C18 phases optimal performance with acidic, basic and chelating compounds. when being used for separations of hydrophobic compounds with typical reversed-phase eluents. They also show stable base lines Analysis of Vitamin B and high sensitivity even under neutral pH conditions and without buffer or counter-ion additives. 100 Column: DAISOGEL SP-120-5-ODS-RPS 150 x 6 mm Eluent: 10 mM sodium phosphate (pH 5.0) /CH3CN (85/15) Analysis of Pyridine/Phenol Flow Rate: 1.0 mL/min Detection: UV @ 254 nm No change in selectivity or retention after 100 hours of washing 1 Temperature: 40 °C Compounds: 0 hours 100 hours 50 2 1. Thiamin hydrochloride (Vitamin B1) mV 2. Riboflavin (Vitamin B2) after 0 hrs 100 hrs Pyridine Kpy 0.704 0.686 Phenol Kph 2.129 2.080 Separation α 3.024 3.032 0 4 4 0 5 10 15 20 25 min Analysis of Organic Acids Analysis of Antioxidants Showing good retention of ascorbic acid 250 2 Column: DAISOGEL SP-120-5-ODS-RPS 150 x 6 mm 5 Eluent: 30 mM sodium phosphate 3 1 (pH 2.5 with H3PO4) 200 Flow Rate: 1.0 mL/min Column: DAISOGEL SP-120-5-ODS-BP Detection: UV @ 210 nm 150 x 6 mm 1 Temperature: 40 °C 2 Eluent: 0.1 M phosphate buffer (pH 3.1) 150 Flow Rate: 1.0 mL/min Compounds: Detection: ECD 1. Malonic acid Temperature: Ambient mV 2. Nortriptyline 3. Acetic acid 100 Compounds: 4 4. Lactic acid 1. Ascorbic acid 5. Citric acid 2. GSH 6 6. Succinic acid 50 0 0 2.5 5.0 7.5 10 min 0 5 10 min [6] DAISOGEL SP-200-C8-P . Silica-based 200Å C8 phase . Recommended for highly hydrophobic samples C8-P packing materials are bonded with octyl groups and are fully endcapped. They are recommended for compounds which are too strongly retained on C18 phases. 200Å material is widely used for larger molecules with higher hydrophobicity. Selectivity Comparison of Three Daisogel Phases 2 Mobile Phase: 65% MeOH /35% 20 mM SP-120-5-ODS-RPS 1 3 5 K2HPO4 buffer, pH 7.0 Temperature: 23.4 °C 4 6 7 Compounds: 2 1. Uracil 1 3 5 SP-120-5-ODS-BP 4 2. Propranolol 6 7 3. Butylparaben 4. Naphthalene 2 3 5 5. Dipropylphthalate 4 SP-200-5-C8-P 6. Acenaphthene 1 6 7 7. Amitriptyline DAISOGEL SP-120-P Analysis of Fat-Soluble Vitamins under . Ultra-high purity 120Å silica Normal Phase Conditions Column: DAISOGEL SP-120-5-P P phases are based on spherical and totally porous silica gel with metal 3 150 x 6 mm 400 impurities specified to be less than 10 ppm each for Al, Fe, Ti and Zr. Eluent: n-Hexane/CHCl3 (60/40) 2 Flow Rate: 1.0 mL/min Due to its narrow particle size distribution, P series packing material for Detection: UV @ 254 nm Temperature: Ambient preparative chromatography enables higher column efficiencies, lower operating pressure and longer column lifetimes. Compounds: 1. Vitamin A Palmitate Using DAISOGEL, scale-up is guaranteed by using the same production 2. Vitamin K1 mV procedure for all grades. 200 3. Vitamin E 4 4. Vitamin K3 P series packing materials for preparative use are available in 10 µm, 5. Vitamin D 15 µm and 20 µm. High purity 40-60 µm spherical silica is a cost 5 effective alternative for medium and low pressure chromatography. 1 Measurement of Pressure Drop 15 SP-120-20-P 0 SP-120-15-P 0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 min SP-120-10-P 12 ) /m 9 pa 6 Preparative grades packing material in the past have had a wide particle distribution. The advantage of P Pressure (M series packing material for preparative chromatography is that it enables higher column efficiencies and 3 lower operating pressure due to its narrow particle size distribution. 0 0 10 20 30 40 Linear Velocity (cm/min) Condition: MeOH 100% (40 °C) [7] DAISOGEL SP-300-BIO .