DAISOGEL™DAISOGEL™

Packing Materials for Process Purification

[ Process Purification ]

- offered by Waters

DAISOGELPacking Material for Process Purification

[3]

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

[4]

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

lifetimes often result in lost samples, repeat purification runs and poor

scalability from small to larger volume columns. Common HPLC prep

column failures are due to mechanical instabilities, especially voids in the

packed bed. The patented OBD Technology1 combines optimized packing

processes with a unique set of column designs ensuring increased lifetime

and better reproducibility.

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

dependent upon the specific properties of the chromatographic particles

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.

Optimized Bed Density is achieved by applying pre-determined axial force only at the column inlet.

OBD Optimized Bed Density

High-pressure Slurry PackingLocalized Bed Expansion Near Inlet

Axial-ram Slurry PackingLocalized Compression Near Inlet

OBD ProcessOptimum Bed Density Throughout

[5]

DAISO CO., LTD.

DAISO CO., LTD., headquartered in Osaka,

Japan, is a technology-oriented chemical

company supporting industries including

medicine, agriculture, IT, machinery,

textiles, and construction through

innovative products.

PrODuCt Offering inCLuDeS:

Daisogel packing materials

Pharmaceutical intermediates

Fine chemicals

Proprietary functional chemicals

soda products and Chlor-alkali

synthetic and fabricated resin products

www.daiso-co.com

DaiSOgeL PaCking materiaL

DAISO CO., LTD. is one of the leading silica gel manufacturers for

liquid chromatography, using inorganic synthesis technology. Since

1992, DAISO has added large-scale manufacturing capability for

DAISOGEL media to fully support the process purification laboratories

worldwide. DAISOGEL phases are used for various applications

including medicines and pharmaceuticals, agricultural chemicals,

food ingredients, biomolecules, liquid crystals and functional dyes.

DAISOGEL packing materials have been designed to meet the needs of

process purification. These speherical silica-based packing materials

allow for high mass loading and maximum resolution which can result

in reducing the number of repeated injections. Through high density

bonding and endcapping, DAISOGEL packing materials have shown

extended life and durability. These benefits have helped customers in

reducing their overall purification costs.

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

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.

[6]

DaiSOgeL SP-120-ODS-rPS

Silica-based 120Å C18 phase

High coverage and exhaustive endcapping

ODS-RPS packings featuring maximum surface coverage are the ideal choice for a

wide variety of organic compounds. Carefully controlled full endcapping leads to

optimal performance with acidic, basic and chelating compounds.

DAISOGEL Packing Materials

DaiSOgeL SP-120-ODS-BP

Silica-based 120Å C18 phase

Suitable for hydrophilic compounds separation

Longer lifetime in aqueous eluents

Different selectivity from ODS-RPS

ODS-BP phases are designed to show extended selectivity for

hydrophilic and polar compounds which are either poorly or not

retained on other phases. Typical applications are separations of

biomolecules and metabolites such as oligosaccharides, amino

acids, small peptides, nucleotides and organic acids. They are

endcapped and show similar selectivity as conventional C18 phases

when being used for separations of hydrophobic compounds with

typical reversed-phase eluents. They also show stable base lines

and high sensitivity even under neutral pH conditions and without

buffer or counter-ion additives.

0 5 10 15 20 25 min

0

50

100

mV

1

2

Column: DAISOGEL SP-120-5-ODS-RPS 150 x 6 mmEluent: 10 mM sodium phosphate (pH 5.0) /CH3CN (85/15)Flow Rate: 1.0 mL/minDetection: UV @ 254 nmTemperature: 40 °C

Compounds: 1. Thiamin hydrochloride (Vitamin B1)2. Riboflavin (Vitamin B2)

analysis of vitamin B

0 2.5 5.0 7.5 10 min

0

50

100

150

200

250

mV

1

2

3

4

5

6

Column: DAISOGEL SP-120-5-ODS-RPS 150 x 6 mmEluent: 30 mM sodium phosphate (pH 2.5 with H3PO4)Flow Rate: 1.0 mL/minDetection: UV @ 210 nmTemperature: 40 °C

Compounds: 1. Malonic acid2. Nortriptyline3. Acetic acid4. Lactic acid5. Citric acid6. Succinic acid

analysis of Organic acids

0 hours 100 hours

4 4

0 hrsafter

100 hrsPyridine Kpy 0.704 0.686Phenol Kph 2.129 2.080Separation α 3.024 3.032

analysis of Pyridine/Phenol No change in selectivity or retention after 100 hours of washing

0 5 10 min

1

2

Column: DAISOGEL SP-120-5-ODS-BP 150 x 6 mmEluent: 0.1 M phosphate buffer (pH 3.1)Flow Rate: 1.0 mL/minDetection: ECDTemperature: Ambient

Compounds: 1. Ascorbic acid2. GSH

analysis of antioxidants Showing good retention of ascorbic acid

DaiSOgeL SP-120-P

Ultra-high purity 120Å silica

P phases are based on spherical and totally porous silica gel with metal

impurities specified to be less than 10 ppm each for Al, Fe, Ti and Zr.

Due to its narrow particle size distribution, P series packing material for

preparative chromatography enables higher column efficiencies, lower

operating pressure and longer column lifetimes.

Using DAISOGEL, scale-up is guaranteed by using the same production

procedure for all grades.

P series packing materials for preparative use are available in 10 µm,

15 µm and 20 µm. High purity 40-60 µm spherical silica is a cost

effective alternative for medium and low pressure chromatography.

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.

analysis of fat-Soluble vitamins under normal Phase Conditions

SP-120-5-ODS-RPS

SP-120-5-ODS-BP

SP-200-5-C8-P

1

2 3

4

5

6 7

1

2 3

4 5

6 7

1

2 3 4

5

6 7

Mobile Phase: 65% MeOH /35% 20 mM K2HPO4 buffer, pH 7.0Temperature: 23.4 °C

Compounds:1. Uracil2. Propranolol3. Butylparaben4. Naphthalene5. Dipropylphthalate6. Acenaphthene7. Amitriptyline

Selectivity Comparison of three Daisogel Phases

Preparative grades packing material in the past have had a wide particle distribution. The advantage of P series packing material for preparative chromatography is that it enables higher column efficiencies and lower operating pressure due to its narrow particle size distribution.

15

12

9

6

3

00 10 20 30 40

Linear Velocity (cm/min)Condition: MeOH 100% (40 °C)

Pres

sure

(Mpa

/m)

Measurement of Pressure Drop

SP-120-20-PSP-120-15-PSP-120-10-P

400

200

0

mV

1

2

3

4

5

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

Column: DAISOGEL SP-120-5-P 150 x 6 mmEluent: n-Hexane/CHCl3 (60/40)Flow Rate: 1.0 mL/minDetection: UV @ 254 nmTemperature: Ambient

Compounds: 1. Vitamin A Palmitate2. Vitamin K13. Vitamin E4. Vitamin K35. Vitamin D

[7]

[8]

excellent Durability in acidic environment

accelerated acidic Duration test Condition

Mobile Phase: CH3CN/1.0% TFA aqueous (10/90), pH 1.0Temperature: 70 °CPurge Time: 3 hr

Chromatographic Test Condition

Mobile Phase: CH3OH/H2O (35/65)Temperature: 40 °CDetection: UV @ 254 nm

Compounds: 1. Uracil 2. Methyl Benzoate 3. Toluene4. Naphthalene

Purge Time (hr)

Rete

ntei

on to

Initi

al (%

)

0 3 6 9 12 15 18 210

20

40

60

80

100

24

Competitor A (300Å, C4)Competitor B (300Å, C4)

DAISOGEL SP-300-C4-BIO

DAISOGEL SP-300-C4-BIO

20 min0 2 4 6 8 10 12 14 16 18

250 CV Purged

Initial

1 2

3

4

Competitor B (300Å, C4)

20 min0 2 4 6 8 10 12 14 16 18

250 CV Purged

Initial

2

34

1

Competitor A (300Å, C4)

20 min0 2 4 6 8 10 12 14 16 18

250 CV Purged

Initial

2

34

1

DaiSOgeL SP-300-BiO

Suitable for bulky biomolecules, proteins

300Å pore size, narrow particle size distribution, ultra high purity totally spherical silica gel

High density bonding for extreme performance

Available types: SP-300-ODS-BIO, SP-300-C4-BIO

The improved high density bonding and full endcapping make it most suitable for general use and to separate or purify high molecular weight

compounds, especially proteins. Because of significant improvement in durability, these novel phases exhibit excellent acidic and alkaline resistance.

These can be used for an extended period of time under acidic mobile-phase condition and rinsed for regeneration with NaOH containing solvents.

The C4 phase is recommended for compounds too strongly retained on ODS C18 phases.

[9]

accelerated alkaline Duration test Condition

Mobile Phase: CH3CN/0.01 NaOH aqueous (10/90), pH 12Temperature: 25 °CPurge Time: 3 hr

Chromatographic Test Condition

Mobile Phase: CH3OH/H2O (35/65)Temperature: 40 °CDetection: UV @ 254 nm

Compounds: 1. Uracil 2. Methyl Benzoate 3. Toluene 4. Naphthalene

excellent Durability in alkaline environment

0 2 4 6 8 10 12 14 16 18 20 min0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 min

DAISOGEL SP-300-10-C4-BIO Competitor C (300Å, 15/20 µm, C4)

Initial

After 76 hr (25 cycles)

Initial

After 19 hr (6 cycles)

After 18 hr (6 cycles)

43

214

3

2

1

Purge Time (hr)

Rete

ntei

on to

Initi

al (%

)

0 10 20 30 40 50 60 70 8050

60

70

80

90

100

110

Daisogel SP-300-10-C4-BIO76 hours: 68.8%

Competitor C (300Å, 15 µm, C4)18 hours: 66.6%

p ( , µ , 4)

[10]

Ordering Information

Product Name Pore Size (Å) Particle Size (µm) Pore Volume (mL/g) Surface Area (m2/g) % Carbon Endcapping

SP-120-ODS-RPS 120 5, 10, 15, 20, 40/60 1.0 300 17 YesSP-120-ODS-BP 120 5, 10, 15, 20, 40/60 1.0 300 15 YesSP-200-C8-P 200 5, 10, 15, 20, 40/60 1.1 200 7 YesSP-120-P 120 5, 10, 15, 20, 40/60 1.0 300 N/A N/ASP-300-ODS-BIO 300 5, 10, 15, 20 0.9 100 8 YesSP-300-C4-BIO 300 5, 10, 15, 20 0.9 100 3 Yes

DaiSOgeL PHySiCaL PrOPertieS

Silica particle shape: SP (spherical)

Pore size (Å): 120, 200, 300

Particle size (µm): 5, 10, 15, 20, 40/60

Bonded phase: see table

SP- 200- P

Base silica purity: P indicates ultra-high purity; Bio indicates ultra-high purity

high acidic resistance; suitable for organic compound separation

low carbon content; suitable for hydrophilic and polar compound separation in up to 100% aqueous eluents

for compounds too strongly retained on ODS phases

if no bonded phase is indicated, the product is bare silica

designed for biopharmaceutical applications; novel bonding technique ensures strong resistance against alkalic and acidic conditions

new variation of the C4 phase; novel bonding technique ensures strong resistance against alkalic and acidic conditions

5- C8-

guiDe tO interPreting DaiSOgeL PrODuCt nameS

narrow particle size distribution

exceptional batch-to-batch reproducibility

enhanced mechanical stability

Suitable for Dynamic axial Compression columns

DaiSOgeL PrODuCt CHaraCteriStiCS

[11]

DaiSOgeL PaCking materiaLS

Particle Size DimensionSP-120- ODS-RPS

SP-120- ODS-BP

SP-200- C8-P

SP-120-PSP-300- ODS-BIO

SP-300- C4-BIO

5 µm 4.6 x 100 mm 186005201 186005171 186005156 186005186 186005141 1860051265 µm 4.6 x 150 mm 186005202 186005172 186005157 186005187 186005142 1860051275 µm 4.6 x 250 mm 186005203 186005173 186005158 186005188 186005143 186005128

10 µm 4.6 x 100 mm 186005204 186005174 186005159 186005189 186005144 18600512910 µm 4.6 x 150 mm 186005205 186005175 186005160 186005190 186005145 18600513010 µm 4.6 x 250 mm 186005206 186005176 186005161 186005191 186005146 18600513110 µm OBD 19 x 100 mm 186005207 186005177 186005162 186005192 186005147 18600513210 µm OBD 19 x 150 mm 186005208 186005178 186005163 186005193 186005148 18600513310 µm OBD 19 x 250 mm 186005209 186005179 186005164 186005194 186005149 18600513410 µm OBD 30 x 100 mm 186005210 186005180 186005165 186005195 186005150 18600513510 µm OBD 30 x 150 mm 186005211 186005181 186005166 186005196 186005151 18600513610 µm OBD 30 x 250 mm 186005212 186005182 186005167 186005197 186005152 18600513710 µm OBD 50 x 100 mm 186005213 186005183 186005168 186005198 186005153 18600513810 µm OBD 50 x 150 mm 186005214 186005184 186005169 186005199 186005154 18600513910 µm OBD 50 x 250 mm 186005215 186005185 186005170 186005200 186005155 186005140

DaiSOgeL COLumnS PaCkeD By WaterS

SP-120-ODS-RPS

Qty 10 µm 15 µm 20 µm 40/60 µm

100 g 186005014 186005050 186005086 -500 g 186005015 186005051 186005087 1860051171 kg 186005016 186005052 186005088 1860051185 kg 186005017 186005053 186005089 186005119

10 kg 186005018 186005054 186005090 18600512020 kg 186005019 186005055 186005091 186005121

SP-120-ODS-BP

Qty 10 µm 15 µm 20 µm 40/60 µm

100 g 186005002 186005038 186005074 -500 g 186005003 186005039 186005075 1860051051 kg 186005004 186005040 186005076 1860051065 kg 186005005 186005041 186005077 186005107

10 kg 186005006 186005042 186005078 18600510820 kg 186005007 186005043 186005079 186005109

SP-200-C8-P

Qty 10 µm 15 µm 20 µm 40/60 µm

100 g 186004996 186005032 186005068 -500 g 186004997 186005033 186005069 1860050991 kg 186004998 186005034 186005070 1860051005 kg 186004999 186005035 186005071 186005101

10 kg 186005000 186005036 186005072 18600510220 kg 186005001 186005037 186005073 186005103

SP-120-P

Qty 10 µm 15 µm 20 µm 40/60 µm

100 g 186005008 186005044 186005080 -500 g 186005009 186005045 186005081 1860051111 kg 186005010 186005046 186005082 1860051125 kg 186005011 186005047 186005083 186005113

10 kg 186005012 186005048 186005084 18600511420 kg 186005013 186005049 186005085 186005115

SP-300-ODS-BIO

Qty 10 µm 15 µm 20 µm

100 g 186004990 186005026 186005062500 g 186004991 186005027 1860050631 kg 186004992 186005028 1860050645 kg 186004993 186005029 186005065

10 kg 186004994 186005030 18600506620 kg 186004995 186005031 186005067

SP-300-C4-BIO

Qty 10 µm 15 µm 20 µm

100 g 186004984 186005020 186005056500 g 186004985 186005021 1860050571 kg 186004986 186005022 1860050585 kg 186004987 186005023 186005059

10 kg 186004988 186005024 18600506020 kg 186004989 186005025 186005061

For additional DAISOGEL products, please contact your local Waters sales representative.

© 2010 Waters Corporation. Waters, OBD and The Science of What’s Possible are

trademarks of Waters Corporation. DAISOGEL is a trademark of DAISO CO., LTD.

720003321EN May 2010 VW-KK-FP

The quality management system of Waters’ manufacturing facilitiesin Taunton, Massachusetts and Wexford, Ireland complies with the Interna-tional Standard ISO 9001:2000 Quality Management and Quality Assurance Standards. Waters’ quality management system is periodically audited by the registering body to ensure compliance.

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