Mise en page 1 (Page 1) - SocochimHEMA 40, 10 µm 101100005 HEMA 100, 10 µm 101120005 HEMA-S 300, 10 µm 101141005 HEMA-S 1000, 10 µm 101161005 Column STEEL 8x250 mm Cat. Number

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Distributed by

Sorbents and Columns HPLC and SPE

Gallery Science 206 Limestone Dr

Bellefonte PA 16823

Phone: 814-360-0130 Fax: 209-254-9501

www.galleryscience.com

Contents Page

General ....................................................................................................................................................... 2 Introduction ................................................................................................................................................. 2 Ordering information ................................................................................................................................... 2 Catalogue numbers ..................................................................................................................................... 2 Shipping Conditions .................................................................................................................................... 2 Terms of Payment ....................................................................................................................................... 2 Return of Products ...................................................................................................................................... 2 Quality standards ........................................................................................................................................ 2 Warranty ....................................................................................................................................................... 2 Hydroxyethyl Methacrylate Sorbents - HEMA ......................................................................................... 4 Size Exclusion Chromatography .................................................................................................................. 4 Ion Exchange Chromatography ................................................................................................................... 6 Affinity Supports ........................................................................................................................................... 8 Reversed Phase Chromatography ............................................................................................................... 9 Hydrophobic Interaction Chromatography ................................................................................................ 11 Ion Chromatography ................................................................................................................................. 12 Column for Isolation of PCBs from Fats .................................................................................................... 14 Ion Exclusion Chromatography Columns .................................................................................................. 15 Spherical Silica Sorbents ......................................................................................................................... 19 Normal Phase Chromatography .............................................................................................................. 19 Reversed Phase Chromatography ........................................................................................................... 21 Ion Exchange Chromatography ............................................................................................................... 23 Macroporous Silica Sorbents .................................................................................................................... 24 Polymeric Sorbents for Gas Chromatography .................................................................................... 25 Packed Columns ....................................................................................................................................... 26 Stainless Steel Analytical and Semipreparative Columns ........................................................................ 26 Stainless Steel Preparative Columns ........................................................................................................ 26 Inert PEEK Analytical and Semipreparative Columns .............................................................................. 26 Inert Titanium Analytical and Semipreparative Columns .......................................................................... 26 Compact Glass Cartridge (CGC) System ................................................................................................. 27 CGC Column Holders ............................................................................................................................... 27 CGC Guard Columns for Stainless Steel Columns .................................................................................. 28 Plastic Cartridges ...................................................................................................................................... 29 Application Note Listing ......................................................................................................................... 30 Classification of Sorbents, Powders, and Solid Particles .................................................................. 39

General Information

2

Introduction TESSEK’s main product line is HEMA (HydroxyEthylMethAcrylate) sorbents. Originally developed at the Institute of Macromolecular Chemistry in Prague, Czech Republic, in 1970, the technology and all associated patents were assigned to TESSEK at its foundation in 1987. From this basic technology, TESSEK developed a complete line of rigid polymeric HPLC sorbents for ion exchange, size exclusion, hydrophobic interaction, and affinity chromatography, as well as many special purpose separation media. TESSEK also manufactures a complete line of silica-based chromatography sorbents. Both types of sorbents are packed into many formats of stainless steel, biocompatible PEEK, inert Titanium alloy, and Compact Glass Cartridge (CGC) columns. Sorbents

TESSEK SEPARON HEMA sorbents are based on a rigid macroporous and hydrophilic polymer matrix. With an unique combination of stability (mechanical, chemical and biological), perfectly spherical shape of particles, macroporous structure and hydrophilic nature, SEPARON HEMA meets the requirements for a versatile chromatographic sorbent. HEMA sorbents are available in various particle sizes ranging from 7 to 60 µm and with exclusion limits ranging from 40 to 2000 kDa. The HEMA-S matrix modification enhances the rigidity of the sorbent, while the HEMA-BIO matrix modification yields a highly hydrophilic sorbent. Further modifications yield sorbents suitable for ion chromatography or reversed phase chromatography. Matrices with activated supports (e.g., containing epoxy or vinylsulpho groups) for additional "custom made" modifications are also available.

TESSEK SEPARON SGX sorbents are based on a totally porous, spherical silica. The sorbent is prepared by an original technology, ensuring high batch-to-batch reproducibility. It features high sorption capacity, good mechanical strength, enhanced chemical stability, and a wide pore size distribution. The average pore diameter is approx. 8 nm (special macroporous sorbents for size exclusion chromatography have pore diameter 20, 50 and 100 nm respectively), the average surface area 500 m2/g and the pore volume 1 mL/g. The modified sorbents are stable in acidic and neutral medium. SEPARON SGX sorbents are available in various particle sizes ranging from 3 to 60 µm and various modifications suitable for normal, reverse phase, and ion chromatography respectively. HPLC Columns

Stainless steel columns, plastic (PEEK) columns, titanium columns and compact glass cartridges of original construction are packed with these sorbents. Plastic cartridges made from medical grade PP are packed with macroparticulate sorbents for solid phase extraction (SPE). Titanium columns posses high biochemical inertness similar to that of PEEK columns; in addition, they exhibit high mechanical strength and withstand high pressure.

Services

Columns from both TESSEK and other producers can be repacked on request, including preparative columns up to 7” diameter. A minimum order is required for custom-packed products (quantity depends on product type). Contact Gallery Science for more information.

We also offer classification of HPLC sorbents, solid particles, and powders (see Page 39).

Catalog Numbers

Twelve digit catalog numbers are used for all products. The first three digits denote the column type and dimensions. The next six are used for sorbent specification, and the last three digits usually denote pieces per package.

Product Returns

No returns are accepted without prior authorization.

Usage

The products indicated in this catalog are intended for laboratory use only and are not tested for direct clinical, medical, or food use. The purchaser assumes full responsibility for such applications.

Warranty

European standard conditions are guaranteed. Product specifications and prices subject to change without notice.

Ordering Information

When ordering, please provide the following information:

• Catalog number • Product description • Quantity • Shipping and billing addresses • Purchase order number • Telephone/fax/email and name of contact person

General Information

3

Do you use these Metrohm IC columns:

Anion Dual 1 - 150 (6.1006.020) Anion Dual 1 - 30 (6.1006.030) Anion Dual 1 - 70 (6.1006.040) Anion Preconcentration column (6.1006.200) Or compact glass system with holders?

Then you are already familiar with TESSEK’s patented production. Since 1987, TESSEK Ltd. has been producing sorbents based on an inorganic matrix (silica gel, SEPARON SGX) and an organic copolymer (copolymer of 2-hydroxyethyl methacrylate and ethylene glycol dimethacrylate, SEPARON HEMA), respectively. Products include sorbents and columns for both analytical LC and large scale preparations. Until recently they were sold primarily under OEM arrangements. Now in partnership with Gallery Science, we are pleased to offer the same high-quality products for use in: ♦ hospitals and other health service centers ♦ biochemical laboratories ♦ research and testing laboratories

• medicine • hygiene • pharmacy • drugs

♦ food production ♦ quality control ♦ air and water pollution monitoring ♦ agriculture ♦ veterinary medicine ♦ fermentation (e.g., wine production and brewing industry) ♦ university laboratories (analytical chemistry, biochemistry, etc.). Standards: Each TESSEK product is thoroughly tested during manufacture and before dispatch. All HPLC columns are tested individually, and minimum efficiency is guaranteed. Each column is provided with an individual test report and test chromatogram.

EPA 300.1 part 1 and part 2 Determination of inorganic anions in drinking water by ion chromatography (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-1 - DIN 38405 D 19 Water quality - Determination of the dissolved anions fluoride, chloride, nitrite, orthophosphate, bromide, nitrate and sulfate by liquid chromatography (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-2 - DIN 38405 D 20 Water quality - Determination of dissolved anions by liquid chromatography. Determination of bromide, chloride, nitrate, orthophosphate and sulfate in wastewater (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-2 - DIN 38405 D 22 Water quality - Determination of dissolved anions by liquid chromatography. Determination of chromate, iodide, sulfite, thiocyanate and thiosulfate in wastewater (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

Size Exclusion Chromatography – HEMA SEC

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TESSEK SEPARON HEMA BIO Subsequently hydrophilized, the pure mechanism of separations by steric exclusion results in a linear dependence of log Mw vs. Ve for a series of protein standards and more than 90% recovery for most proteins tested. The rigidity of HEMA BIO sorbents allows the use of aqueous as well as organic solvents, including tetrahydrofuran, dimethylacetamide, cadoxene, etc.

8x250 mm HEMA--BIO 1000, 10 µm

Elution volume, mL

Fig. 3. Calibration Curves for Different Standards Column: Steel 8x250 mm HEMA BIO 100, 10 µm Eluent: : water or tetrahydrofuran Flow rate: 1.0 mL/min. Detection: RI or UV 254 nm Polydextrans: D20, glucose, saccharose, raffinose, cextrans 3000, 4500. Pharmacia dextrans: D10, D20, D40, D70, D110, D 150, D 250, D500, D2000. Polymaltotrioses: Showa Denko P5, P10, P 20, P40, P100, P200, P400, P800, Polystyrenes. For preparative separation of proteins, usually they are applicable only for simple mixtures of proteins with enough differing sizes. Best results are achieved when the exclusion limit is selected so that the bigger molecule is excluded and the smaller one is well within the working range. Use of small particles (10-20 µm) is essential in this technique to achieve sufficient efficiency. The main advantage of HEMA BIO sorbents for the ability to recondition the column using 0.1 M NaOH as required by FDA protocols.

Steel 8x250 mm HEMA-BIO 1000, 10 µm

Time, min.

Fig. 4. SEC Separation of Proteins Column: Steel 8x250 mm HEMA BIO 1000, 10 µm Eluent: 0.1 M phosphate buffer, 0.15 M sodium chloride, pH 7.0 Flow rate: 1.0 mL/min. Detection UV 230 nm Injection: 10 µL Peaks: 1) Thyroglobulin 660 kDa, 2) Bovine serum albumin 67 kDa, 3) Cytochrom C 12.4 kDa, 4) Cytidin-5-mono phosphate 0.32 kDa, 5) LMW Impurity

Steel 8x250 mm HEMA-BIO 100 10 µm

Time, min

Fig. 5. SEC Separation of Proteins Column: Steel 8x250 mm HEMA BIO 100, 10 µm Eluent: 0.1 M phosphate buffer, 0.15 M sodium chloride, pH 7.0 Flow rate: 1.0 mL/min. Detection: UV 230 nm Injection: 10 µL Peaks: as in Fig.4

Size Exclusion Chromatography – HEMA SEC

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TESSEK SEPARON HEMA and HEMA-S The underivatized HEMA 40 and 100 or the additionally cross-linked HEMA-S 300 and 1000 sorbents are used mostly in SEC of polysaccharides. Due to its slight hydrophobicity, they can be used for hydrophobic interaction chromatography of proteins. Compared to HEMA BIO series, they offer higher working volumes.

Ordering Information – Bulk Sorbents

The following sorbents are available in 10 g, 100 g, and bulk quantities:

Bulk Sorbent Exclusion Catalog TESSEK Separon Limit Number

kDa 10 µm 60 µm HEMA 40 40 - 80 100005 110003 HEMA 100 80 - 250 120005 130003 HEMA-S 300 250 - 800 141005 HEMA 300 250 - 800 150003 HEMA-S 1000 800 - 2000 161005 HEMA 1000 800 - 2000 170003 HEMA BIO 40 40 - 80 102005 112003 HEMA BIO 100 80 - 250 122005 132003 HEMA BIO 300 250 - 800 142005 152003 HEMA BIO 1000 800 - 2000 162005 172003

Ordering Information – Packed Columns Steel, Titanium, and PEEK columns are packed individually. Each column is provided with test report and chromatogram.

Column STEEL 8x250 mm Cat. Number

HEMA 40, 10 µm 101100005 HEMA 100, 10 µm 101120005 HEMA-S 300, 10 µm 101141005 HEMA-S 1000, 10 µm 101161005

Column STEEL 8x250 mm Cat. Number

HEMA BIO 40; 10 µm 101102005 HEMA BIO 100, 10 µm 101122005 HEMA BIO 300, 10 µm 101142005 HEMA BIO 1000, 10 µm 101162005

Preparative stainless steel columns with 25 or 46 mm internal diameter are available on request.

Column Ti-Zr 8x250 mm Cat. Number

HEMA BIO 40, 10 µm 201102005 HEMA BIO 100, 10 µm 201122005 HEMA BIO 300, 10 µm 201142005 HEMA BIO 1000, 10 µm 201162005

Column PEEK 7.5x300 mm Cat. Number

HEMA BIO 40, 10 µm 302 102005

HEMA BIO 100, 10 µm 302 122005HEMA BIO 300, 10 µm 302 142005 HEMA BIO 1000, 10 µm 302 162005

Ion Exchange Chromatography – HEMA IEC

6

ION EXCHANGE CHROMATOGRAPHY

Time, min.

Fig. 6. Separation of Proteins at High pH Column: CGC 3x150 mm HEMA BIO 1000 Q, 10 µm Eluent: A 10 mM TRIS, pH 11, B 10 mM TRIS + 0.5 M NaCl, pH 11, Gradient: from 0-30% B in 10 min, 30% B for 7 min, 30-100% B in 8 min. Flow rate: 0.5 mL/min. Detection: UV 280 nm Peaks: 1) chymotrypsinogen, 2) chymotrypsin

Time, min.

Fig. 7. Separation of Protein Standard Column: CGC 3x150 mm HEMA BIO 1000 DEAE 10 µm Eluent: A 20 mM TRIS.HCI, pH 7.8, B 20 mM TRIS HCl + 0.5M NaCl, pH 7.8 Gradient: linear from 0 to 100% B in 30 min Flow rate: 0.5 mL/min. Detection: UV 280 nm Peaks: 1) myoglobin, 2) conalbumin, 3) ovalbumin, 4) soybean trypsin inhibitor. The macroporous structure and hydrophilic nature of the TESSEK Separon HEMA BIO matrix are ideal for forming powerful ion exchangers for chromatography of many types of solutes. The HEMA-S 1000 offers higher capacities, while the HEMA BIO 1000 matrix should be chosen for better protein recovery and enhanced rigidity, necessary for example in organic solvents.

Four different ion-exchange modifications of TESSEK Separon HEMA sorbents are available: DEAE Medium basic anion exchanger containing

diethylaminoethyl groups, widely used for the separations of proteins and biopolymers. The sorbent is supplied in chloride form.

Q Strong basic anion exchanger containing

quaternary amino groups, necessary for use in alkaline pH. The sorbent is supplied in chloride form.

CM Weak acidic cation exchanger containing

carboxymethyl groups, applicable for gentle protein separations. The sorbent is supplied in hydrogen form.

SB Strong acidic cation exchanger containing

sulphobutyl groups with wide applications. The sorbent is supplied in sodium form.

Time, min.

Fig. 8. Separation of Protein Standard Column: CGC 3x150 mm HEMA BI0 1000 SB, 10 µm Eluent: A 20 mM Na2HPO4 , pH 6.8, B 20 mM Na2HPO4 + 0.5 M NaCl, pH 6.8 Gradient: linear from 0 to 100% B in 30 min Flow rate: 0.5 mL/min. Detection: UV 280 nm Peaks: 1) myoglobin, 2) ribonuclease A, 3) cytochrom C, 4) lysozym.


7


The following sorbents are available in 10 g, 100 g, and bulk quantities:

Bulk Sorbent Capacity Catalog number TESSEK Separon mmol/g 10 µm 60 µm HEMA-S 1000 DEAE 1.2 - 1.5 161045 HEMA 1000 DEAE 1.2 - 1.5 170043 HEMA 1000 Q 0.8 - 1.2 160095 170093 HEMA-S 1000 CM 0.9 - 1.1 161055 HEMA 1000 CM 0.9 - 1.1 170053 HEMA-S 1000 SB 1.6 - 2.0 161135 HEMA 1000 SB 1.6 - 2.0 170133 HEMA BIO 1000 DEAE 0.8 - 1.2 162045 172043 HEMA BIO 1000 Q 0.4 - 0.8 162095 172093 HEMA BIO 1000 CM 0.4 - 0.8 162055 172053 HEMABIO1000SB 1.2-1.6 162135 172133

Ordering Information – Packed Columns Steel, Titanium, and PEEK columns are packed individually. CGC system glass columns are packed individually, immersed in 0.01% sodium azide, and sealed in aluminum foil bags. Each column is provided with test report and test chromatogram. CGC 3x30 mm columns are immersed in 0.01% sodium azide, and sealed in aluminum foil bags as a set of 3.

Preparative columns with 25 or 46 mm internal diameter are available on request. The sorbents in 60 µm particle size are also available in HEMA-cartridges for solid phase extraction (see page 29).

HEMA BIO 1000 DEAE 10 µm Cat. Number STEEL 8x250 mm 101 162045 STEEL 8x80 mm 109 162045 Ti / Zr 8x250 mm 201 162045 Ti / Zr 8x80 mm 209 162045 PEEK 7.5x250 mm 301 162045 PEEK 7.5x75 mm 304 162045 PEEK 4.6x250 mm 341 162045 PEEK4.6x150mm 347 162045 PEEK 4.6x50mm 351 162045 CGC 3x150 mm 901 162045 CGC 3x30 mm 911 162045

HEMA BIO 1000 Q 10 µm Cat. Number STEEL 8x250 mm 101 162095 STEEL 8x80 mm 109 162095 T1 / Zr 8x250 mm 201 162095 Ti / Zr 8x80 mm 209 162095 PEEK 7.5x250 mm 301 162095 PEEK 7.5x75 mm 304 162095 PEEK 4.6x250 mm 341 162095 PEEK 4.6x150 mm 347 162095 PEEK 4.6x50mm 351 162095 CGC 3x150 mm 901 162095 CGC 3x30 mm 911 162095

HEMA BIO 1000 CM 10 µm Cat. Number STEEL 8x250 mm 101 162055 STEEL 8x80 mm 109 162055 Ti / Zr 8x250 mm 201 162055 Ti / Zr 8x80 mm 209 162055 PEEK 7.5x250 mm 301 162055 PEEK 7.5x75 mm 304 162055 PEEK 4.6x250 mm 341 162055 PEEK 4.6x150 mm 347 162055 PEEK 4.6x50mm 351 162055 CGC 3x150 mm 901 162055 CGC 3x30 mm 911 162055

HEMA BIO 1000 SB 10 µm Cat. Number STEEL 8x250 mm 101 162135 STEEL 8x80 mm 109 162135 Ti / Zr 8x250 mm 201 162135 Ti / Zr 8x80mm 209 162135 PEEK 7.5x250 mm 301 162135 PEEK 7.5x75 mm 304 162135 PEEK 4.6x250 mm 341 162135 PEEK 4.6x150 mm 347 162135 PEEK 4.6x50mm 351 162135 CGC 3x150 mm 901 162135 CGC 3x30 mm 911 162135

Spherical Silica Sorbents – Reversed Phase Chromatography

8

AFFINITY SUPPORTS The high density of hydroxyl groups on the internal surface provides TESSEK Separon HEMA with hydrophilic properties and excellent derivatization possibilities for the preparation of different chromatographic sorbents. Common activation techniques, e.g., cyanogen bromide, triazine, and others, can be used for ligand immobilization on TESSEK Separon HEMA materials. However, those techniques are rather laborious and usually present health hazards. To overcome this step, TESSEK offers ready-to-use activated affinity supports with a long shelf life. Two different modifications with different levels of activation are available: the epoxy (E, E-L, E-H) and vinyl sulphone (VS, VS-L). The suitable level of activation depends on the type of ligand. For the binding of low molecular weight ligands to high surface density, a high level of activation should be used. For macromolecular ligands, especially where biological activity should be retained, a low activation level is necessary. Epoxy-activated supports react under suitable conditions with a variety of functional groups, especially with amino-, thio-, carboxy-, and hydroxy groups. Vinyl sulphone-activated supports exhibit higher reactivity, particularly for hydroxy groups. They are especially recommended for immobilization of large proteins and active enzymes. Recommended procedure of ligands immobilization to Separon HEMA activated support is a simple five-step process: GENERAL PROCEDURE

1. Equilibrate the column with the coupling buffer. 2. Add ligand dissolved in the coupling buffer and let

the reaction proceed for the time required (e.g., stop-flow mode, recycling mode).

3. Wash the column with several column volumes of distilled water to remove excess of unbound ligand.

4. Add blocking reagent and let the blocking reaction proceed for the time required (e.g., stop-flow mode, recycling mode).

5. Wash the column again with several column volumes of distilled water, followed by multiple volumes of buffers. The column is now ready for use.

FACTORS INFLUENCING THE COUPLING REACTION For a successful immobilization of the ligand to Separon HEMA activated supports, several factors must be considered, the most important ones being the ligand properties and the desired effect. pH: The E and VS groups read with amino-, hydroxy-,

phenol-, mercapto-, carboxy-, imidazole-, and indole functional groups. Of those, the amino groups are the most reactive and useful ones for ligand coupling. As the slightly alkaline pH was found to be optimal for ligands containing amino groups, a buffer with pH 9 is a good starting point.

Reaction time: A common characteristic of ligand incorporation is a rapid increase during the first few hours, and after this, the rate levels off. A logarithmic function is usually a good approximation for this dependence.

Ionic strength: The reaction depends on the surface concentration of the ligand. The presence of a high salt concentration (ammonium sulphate) can force the ligand molecules out of the solution (salting out effect) and thus increase the reaction yield.

Temperature: Increased temperatures will speed up the coupling reaction, however, ambient or lower temperatures should be used for immobilization of thermally labile ligands.

Activation: For maximum bonding of low molecular ligands, the highest level of activation should be chosen. For lower loadings, a lower activation or shortened reaction time is applicable. For the bonding of macromolecular ligands, especially when the biological activity should be retained (enzymes etc.), the low activation level is recommended.

Blocking of excessive E or VS groups: The excess of unreacted E or VS groups should be blocked before affinity chromatography can be performed. The blocking procedure selected depends mainly on the type and stability of the ligand. The most common procedure apply 0.1 M ethanolamine as a blocking reagent.


The following sorbents are available in 10 g, 100 g, and bulk quantities. Due to the limited shelf life, no HPLC columns are available. They can be packed on request, but must be used within three weeks from production. Plastic cartridges HEMA-cart (Cat. No. 421) are packed with dried sorbents (see page 29).


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TESSEK Separon Capacity Catalog number mmol/g 10 µm 60 µm

HEMA 1000 E 0.7 - 0.9 160015 170013 HEMA-S 1000 E-L 0.1 - 0.3 161025 HEMA 1000 E-L 0.1 - 0.3 170023 HEMA 1000 E-H min 1.4 160085 170083 HEMA BIO 1000 E 0.5 - 0.7 162015 172013 HEMA BIO 1000 E-L 0.05 - 0.2 162025 172023 HEMA BIO 1000 E-H 0.9 - 1.2 162085 172083 HEMA BIO 1000 VS 0.01 - 0.05 162125 172123 HEMA BIO 1000 VS-L 0.005 - 0.010 162115 172113

REVERSED PHASE CHROMATOGRAPHY

Time, min.

Fig. 11. Separation of Amines Column: CGC 3x150 mm HEMA-S 1000 C18, 10 µm Eluent: methanol-water 70:30 Flow rate: 0.5 mL/min. Detection UV 254 Injection: 20 µL Peaks: 1) aniline, 2) pyridine, 3) 2.5-dimethylaniline, 4) 4-chloroaniline. The rigidity of the matrix ensures that the columns can be operated with pressures up to 100 bar (1450 psi) and that the efficiency is not affected by changes in the mobile phase composition. The lack of silanol groups and the outstanding stability of TESSEK Separon HEMA BIO 1000 C18 makes it especially suited for separations of basic compounds and expand the application area for reversed phase chromatography. Most problems connected with the use of silica-based sorbents in reversed phase chromatography are associated with the presence of residual silanol groups and limited stability of the matrix under alkaline conditions. Polymer based TESSEK Separon HEMA reversed phase sorbents were developed with the

purpose of creating efficient and highly stable reversed phase material. The new sorbent is produced by grafting the macroporous hydrophilic TESSEK Separon HEMA 1000 co-polymer with C18 alkyl chains.

The newly used HEMA BIO matrix offers enhanced rigidity with essentially the same selectivity. The sorbent is stable in the pH range 2-12 and is compatible with all commonly used chromatography solvents. The wide-pore structure of the TESSEK Separon HEMA BIO 1000 C18 matrix allows separations of macromolecules (peptides, proteins, polynucleotides, etc.).

Time, min.

Fig. 12. Separation of Protein Standard. Column: PEEK 4.6x100 mm HEMA-S 1000 C18, 10 µm Gradient 20- 64% acetonitrile in 0.1% trifiuoroacetic acid in 40 min. Detection: UV 280 nm Injection: 20 µL Flow rate: 1.0 mL/min. Peaks: 1) ribonuclease A, 2) cytochrom C, 3) alpha-Lactalbumin, 4) myoglobin (ape), 5) hem (myoglobin)


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Ordering Information – Bulk Sorbents The following sorbents are available in 10 g, 100 g, and bulk quantities:

Catalog number Bulk Sorbent TESSEK Separon

Functional group

10 µm 60 µm

HEMA-S 1000 C18 HEMA 1000 C18 HEMA-S 1000 C8 HEMA-S 1000 C4

octadecyl octadecyl

octyl butyl

161155

161215 161165

170153

HEMA BIO 1000 C18 octadecyl 162155 172153

Packed Columns

Steel, Titanium and PEEK columns are packed individually. CGC system glass columns are packed individually, immersed in 0.01% sodium azide, and sealed in aluminum foil bags. Each column is provided with test report and test chromatogram. CGC 3x30 mm columns are immersed in 0.01% sodium azide and sealed in aluminum foil bags as a set of 3. Preparative columns with 25 or 46 mm internal diameter are available on request. Sorbents in 60 µm particle size are also available in HEMA-cart cartridges for solid phase extraction (see page 29).

Ordering Information – Packed Columns HEMA-S 1000 C18 10 µm Column Cat. Number

STEEL 8x250 mm 101 161155 STEEL 8x80 mm 109 161155 Ti / Zr 8x250 mm 201 161155 Ti / Zr 8x80 mm 209 161155 PEEK 7.5x250 mm 301 161155 PEEK 7.5x75 mm 304 161155 PEEK 4.6x250 mm 341 161155 PEEK 4.6x150 mm 347 161155 PEEK 4.6x50 mm 351 161155 CGC 3x150 mm 901 161155 CGC 3x30 mm 911 161155

HEMA-S 1000 C4, 10 µm Column

Cat. Number


HEMA-S 1000 C8, 10 µm Column

Cat. Number


HEMA BIO 1000 C18, 10 µm Column

Cat. Number

STEEL 8x250 mm 101 162155

STEEL 8x80 mm 109 162155 Ti / Zr 8x250 mm 201 162155 Ti / Zr 8x80 mm 209 162155 PEEK 7.5x250 mm 301 162155 PEEK 7.5x75 mm 304 162155 PEEK 4.6x250 mm 341 162155 PEEK 4.6x150 mm 347 162155 PEEK 4.6x50 mm 351 162155 CGC 3x150 mm 901 162155 CGC 3x30 mm 911 162155


11

HYDROPHOBIC INTERACTION CHROMATOGRAPHY Hydrophobic interaction chromatography is one of separation methods widely applied to separation of proteins. In contrast to reversed phase chromatography, the separation under nondenaturing conditions allows isolation of highly active proteins from complex natural mixtures. The method is based on elution of proteins from slightly hydrophobic sorbent by descending gradient of ionic strength, i.e., ammonium sulfate in phosphate buffer. The slightly hydrophobic nature of hydroxyethyl methacrylate gel TESSEK Separon HEMA-S 1000 allows its direct use for hydrophobic interaction chromatography. However, the best results are obtained on specially developed TESSEK Separon HEMA BIO 1000 Phenyl. This sorbent contains approx 0.1 mmol/g of phenyl groups attached to a highly hydrophilic surface of HEMA BIO 1000 matrix.

Time, min.

Fig. 13. Separation of Proteins by HIC. Column: Steel 8x80 mm HEMA BIO 1000 Phenyl 10 µm. Eluent: A-2.2 M ammonium sulfate, 0.1 M phosphate, pH 7.0, B-0.1 M phosphate, pH 7.0, Gradient: from 0 to 100% B in 30 min Flow rate: 1.0 mL/min. Detection: UV 280 nm Injection: 250 µL Peaks: 1) Myoglobin 1 mg/mL, 2) Ribonuclease A 1mg/mL, 3)Lysozym 0.4 mg/mL, 4) Chymotrypsin 1 mg/mL

Time, min.

Fig. 14. Separation of Proteins by HIC. Column: Steel 8x80 mm HEMA BIO 1000 Phenyl 10 µm. Eluent A-2.2 M ammonium sulfate, 0.1 M phosphate, pH 7.0, B-0.1 M phosphate, pH 7. Gradient from 20 to 100% B in 30 min Flow rate: 1.0mI/min Detection: UV 280 nm Injection: 250 µL Peaks: 1)Bovine serum albumin 4.8 mg/mL, 2) Bovine immunoglobulin 2 mg/mL. Ordering Information – Bulk Sorbent The sorbent is available in 10 g, 100 g, and bulk quantities.

Tessek Separon Bulk Sorbent Cat. Number

HEMA BIO 1000 Phenyl 10 µm 162175

Ordering Information – Packed Columns

Preparative columns with standard internal diameters and lengths are available on request. Steel, Titanium, and PEEK columns are packed individually. CGC system glass columns are packed individually, immersed in 0.01% sodium azide, and sealed in aluminum foil bags. A test report and test chromatogram are provided with each column. CGC 3x30 columns are packed as a set of 3.

HEMA BIO 1000 Phenyl 10 µm Column

Cat. Number

STEEL 8x80 mm 109 162175 Ti-Zr 8x80 mm 209 162175 PEEK 7.5x75 mm 304 162175 PEEK 4.6x150 mm 347 162175 PEEK 4.6x50 mm 351 162175 CGC 3x150 mm 901 162175 CGC 3x30 mm 911 162175

Ion Chromatography – HEMA IEC

12

ION CHROMATOGRAPHY

Ion chromatography is a long-established, progressive method for the analysis of different mixtures of anions. When introduced in the late seventies, rather special and complicated equipment was necessary for performing ion chromatography (at that time, meaning a method using conductometric detection with chemically suppressed eluent conductivity). Since then, the range of applications included in the “ion-exchange” category has broadened considerably.

Time, min.

Fig. 15. Separation of Inorganic Anions Standard. Column: CGC 3x150 mm HEMA-S Q-L10 µm Eluent: 1.0 mM potassium hydrogenphthalate, pH 6.0 Flow rate: 0.5 mL/min. Detection: indirect UV at 260 nm Injection: 100 µL Sample: standard mix,1) fluoride 5 mg/I, 2) chloride 5 mg/l, 3) nitrite 5 mg/1, 4) bromide 10 mg/l, 5) nitrate 10 mg/l, 6) sulfate 10 mg/I

Time, min.

Fig. 16. Nitrate Determination in Residues after Industrial and Military Explosives. Column: CGC 3x150 HEMA-S 1000 Q-L 10 µm, Eluent: 0.5 mM sodium sulphosalicylate, pH 6.0, Flow rate: 0.5 mL/min., Detection: indirect photometry at 254 nm, 0.08 AUFS, Injection: 10 µL Fig. A. Soil Extract before Explosion. Peaks: 1) hydrogencarbonate, 2) chloride, 3) nitrite, 4) nitrate, 5) sulphate Fig. B. Soil Extract after Explosion. Peaks: as in Figure A.

Many types of detectors (photometric, conductometric, refractometric) can now be used with the indirect method of detection. However, nearly all anions (with the exception of fluoride, chloride, sulphate, phosphates, carbonate, borate and a few others) are detectable directly by UV photometry in the range 200-215 nm. Low capacity anion exchangers are widely used in the ion chromatography of inorganic and organic anions. Columns packed with these sorbents ensure fast, simple, and sensitive determination of the anions using practically any liquid chromatograph. TESSEK introduces a new TESSEK Separon HEMA-S 1000 Q-L and HEMA S/S 1000 Q-L, quaternary amine anion exchanger with a capacity of around 0.1 mmol/g. This sorbent was found to be well suited for all detection modes commonly used in ion chromatography, including suppressed and nonsupressed conductivity, indirect photometry, and others.

Fig. 17. Separation of Inorganic Anions Standard. Column: CGC 3x150 mm HEMA S/S 1000 Q-L 10 µm Eluent: 8 mmol/I phthalic acid, 2% acetonitrile, pH 4.0 (TRIS), Flow rate: 0.5 mL/min. Injection: 100 µL Detection: conductometric Sample: standard mix, 1) fluoride 5 mg/I, 2) chloride 5 mg/I, 3) nitrite 5 mg/I, 4) bromide 10 mg/I, 5) nitrate 10 mg/I, 6) sulfate 10 mg/I.

Ion Chromatography – HEMA IEC

13

Time, min.

Fig. 18. Determination of Anions in Whiskey. Column: CGC 3x150 TESSEK Separon HEMA S/S 1000 Q-L 10 pm, (known as Metrosep anion Dual, Metrohm cat. no 6.1006.020) Eluent: 2.4 mmol/I NaHCO3,2.5 mmol/I Na2CO3, 2% acetone with chemical suppression. Flow rate: 0.5 mL/min. Injection: 100 µL Peaks: 1) chloride, 2) system peak, 3) nitrate, 4) phosphate, 5) malate, 6) sulfate, 7) oxalate.

Ordering Information Packed Columns Steel and PEEK columns are packed individually. CGC system glass columns are packed individually immersed in 0.01% sodium azide and sealed in aluminum foil bags. Each column is provided with test report and test chromatogram. CGC columns are packed as a set of 3.

TESSEK Separon HEMA-S 1000 Q-L 10 µm Column Cat. Number

Stainless steel 8x80 mm 109 161105 PEEK 4.6x150 mm 347 161105 PEEK 4.6x50 mm 351161105 CGC 3x150 mm 901 161105 CGC 3x30 mm 911 161105

Columns with ISO EPA 300.1 part 1 and part 2 Determination of inorganic anions in drinking water by ion chromatography (TESSEK HEMA

S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-1 - DIN 38405 D 19 Water quality - Determination of the dissolved anions fluoride, chloride, nitrite; orthophosphate, bromide, nitrate and sulfate by liquid chromatography (TESSEK HEMA S/S 1000 Q-L3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-2 - DIN 38405 D 20 Water quality - Determination of the dissolved anions by liquid chromatography. Determination of bromide, chloride, nitrate, orthophosphate and sulfate in wastewater (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

ISO 10304-2 - DIN 38405 D 22 Water quality - Determination of the dissolved anions by liquid chromatography. Determination of chromate, iodide, sulfite, thiocyanate and thiosulfate in wastewater (TESSEK HEMA S/S 1000 Q-L 3x150mm CGC, known as Metrosep Dual 1 - 6.1006.020)

TESSEK Separon HEMA S/S 1000 Q-L 10 µm Column TESSEK Cat. Number METROHM Cat. Number

Stainless steel 8x80 mm 109 164105 - PEEK 4.6x150 mm 347 164105 - PEEK 4.6x50 mm 351 164105 - CGC 3x150 mm 901 164105 6.1006.020 CGC 3x30 mm 911 164105 6.1006.030 CGC 3x70 mm 909 164105 6.1006.040

Column for Isolation of PCBs from Fats

14

COLUMN FOR ISOLATION OF PCBs FROM FATS

Polychlorinated biphenyls (PCB's) are ubiquitous environmental pollutants that are strongly accumulated in lipid portion of biological matrices, especially in storage fat. Gas chromatography (GLC/ECD) is employed for the determination of these contaminants. Polycyclic aromatic hydrocarbons (PAHs) are other important group of pollutants, most commonly determined by reversed phase HPLC with fluorimetric detection. Prior to GLC analysis, however, removing of interfering co-extracted compounds has to be carried out. Adsorption chromatography on Florisil, liquid-liquid partition, or solid phase extraction are techniques often used for this purpose. Recently many laboratories have been introducing gel permeation chromatography (GPC) as a clean-up step involved in sample preparation procedure. In comparison with the above methods, GPC possesses many advantages, the main of them is the possibility of multiple use of the same column without any loss of reproducibility of its separation properties. Automation of the cleaning procedure is thus possible. Efficient separation of PCBs and PAHs from fat, oils, and some other compounds extracted from fats and oils can be achieved on styrenedivinylbenzene copolymer Bio Beads S-X3 (BioRad, South Richmond, CA, USA, particle size 37-75 µm) with chloroform mobile phase. Columns packed with this sorbent are expected to find a wide use in all laboratories engaged in analyses of PCBs and PAHs, especially in foodstuffs and many other biological matrices. For other applications (e.g., pesticides in plant material) the column can be prepared on request as well as in other mobile phases.


Packed Columns The column is packed individually with test report and test chromatogram.

Fig. 19. Separation of PCBs from Oils and Fats Column: Steel 8x500 BlO BEADS S-X3 Eluent: chloroform Flow rate: 0.6 mL/min. Detection: RI (fat), GLC/ECD (PBCs in collected fractions)

Column Steel 8x500 mm Cat. Number

BIO BEADS S-X3 102 913003

Ion Exclusion Chromatography Columns

15

ION EXCLUSION CHROMATOGRAPHY COLUMNS HPLC chromatography is an indispensable tool for separating saccharides and organic acids. Several chromatography modes can be successfully employed. In last years, the ion exclusion mechanism is widely employed for those analyses. Highly cross linked, sulfonated polystyrene-divinylbenzene gels in different ionic forms are the most commonly used. TESSEK introduces columns of this type packed with OSTION cation exchange resins in hydrogen, sodium, calcium and lead forms. The resin in hydrogen form is intended for separation of organic acids. Dilute acid (0.01 M sulfuric acid) is recommended as an eluent. Ambient temperature can be used; however, the resolution increases with increased temperature. Detection is most commonly accomplished by UV at 205 - 220 nm. For hydrophobic or aromatic acids, some organic solvent (methanol, acetonitrile) can be added up to 20%. Sugars and alcohols can also be separated with pure water eluent at 80°C and RI detection; however, some oligosaccharides will be hydrolyzed. In ion exclusion mechanism, the selectivity of separation is not influenced significantly by mobile phase. Instead, other ionic forms (sodium, calcium or lead) or degree of cross linking are used for solving special separation problems. Pure water is used as eluent at increased temperature (60 - 80°C). Refractive index detection is necessary. The efficiency of separation increases significantly with decreased flow rate; thus 0.3 - 0.5 mL/min. are recommended for 8x250 mm columns. Maximum pressure is 7 Mpa (1015 psi). The sodium form column offers improved selectivity for sacchrides, and it can tolerate salt containing samples. It is recommended for direct analysis of molasses and other buffered samples. The calcium form column offers improved selectivity for monosaccharides. Sample deionization is recommended, as anions forming insoluble calcium salts will destroy the column selectivity.

Fig. 20. Separation of Organic Acids Standard. Column: 8x250 mm OSTION LG KS 0800 H` form Eluent: 10mM sulfuric acid, Temperature: 80°C Flow rate: 1mL/min. Detection: UV at 210 nm Peaks: 1) Phenylglyoxalic, 2) Mandelic, 3) o-methylhippuric, 4) Hippuric, 5) m-methylhippuric, 6) p-methylhippuric, 7) Benzoic

Time, min.

Fig. 21. Separation of Sugar Mix. Column: 8x250 mm OSTION LG KS 0800 Pb2+form, coupled with two CGC 3x30 mm TESSEK SEPARON HEMA B1O 1000 SB+Q, 10 µm Eluent: water Flow rate: 0.5 mL/min. Temperature: 80° C Detection: RI Peaks: 1) Melezitose, 2) Saccharose, 3) Lactose, 4) Glucose, 5) Xylose, 6) Galactose, 7) Arabinose, 8) Mannose The lead form column offers best selectivity for certain sugars; however, it is very sensitive to sample desalting. On-line or off-line sample deionization is strongly recommended.


16

Elution volumes (mL) of sugars and alcohols on 8x250 OSTION LG KS 0800 in different ionic forms; Temperature: 80°C, Flow rate: 1 mL/min., Detection: RI.

Mobile phase Water Water Water H2SO4 Form Ca2+ Na2+ Pb2+ 10 mmol/L

H+ Melezitose 4.11 3.58 5.02 Raffinose 4.15 3.62 5.22 Trehalose 4.49 3.99 5.61 3.97 Saccharose 4.50 4.07 5.54 Maltose 4.54 4.11 5.86 3.98 Lactose 4.66 4.16 5.99 4.04 Glucose 5.34 5.20 6.61 4.71 Xylose 5.87 5.66 7.16 5.01 Galactose 5.94 5.54 7.54 4.96 Mannose 6.08 5.58 8.93 4.94 Rhamnose 6.10 5.33 8.04 5.29 Fructose 6.71 5.65 9.05 5.01 Arabinose 6.74 6.10 8.18 5.37 Glycerol 8.15 6.33 9.95 6.79 Methanol 8.23 7.49 8.35 9.77 Ethanol 8.39 7.47 8.67 11.13 Isopropyl alcohol 8.43 7.35 8.72 12.32 Ethylene glycol 8.51 7.03 9.74 8.15 Diethylene glycol 9.42 6.45 12.70 8.16 Sorbitol 10.72 5.46 20.82 5.23 Ribose 10.94 6.83 19.39 5.48 Cyclohexanol 19.51 15.37 22.59 27.29 Benzyl alcohol 24.32 20.28 28.73 22.79

Ordering information – Packed Columns

Column, STEEL 8x250 mm Cat. Number OSTION LG KS 4800 H' form 101 908115 OSTION LG KS 0800.Ca".form 101 908135 OSTION LG KS 0800 Pb2 form 101 908155 OSTION LG KS 0800 Na+ form 101 908175

Column, STEEL 4x250 mm Cat. Number

OSTION LG KS 0800 H' form 141 908115

Column, Plastic 9x20 mm Cat. Number HEMA BIO 1000 DEAE 421 172043006 HEMA BIO 1000 Q 421 172093006 HEMA BIO 1000 SB 421 172133006


17

SAMPLE PREPARATION HINTS

Saccharides Saccharides are practically nonretained on all reversed phase and ion exchange sorbents. Depending on the character of interferences, the sample can be passed before analysis through a SILICA-cart C18 cartridge to remove hydrophobic compounds. Passing through coupled HEMA-cart DEAE and HEMA-cart SB cartridges will remove all ionic compounds. Much simpler chromatograms are achieved when analyzing complex matrix samples. Alternatively, two CGC 3x30 mm guard columns packed with HEMA BIO 1000 Q and SB 10 mm sorbents in OH- and H+ ionic forms respectively can be directly coupled to the column using a column coupling union and appropriate cartridge holder.

Organic acids Selective retention of organic acid on anion exchangers can be employed for their isolation and preconcentration from complex mixtures. HEMA-cart DEAE or HEMA cart Q are best suited for this purpose.

Fig. 22. Separation of Sugars in Wine. Sample filtered through HEMA -cart DEAE and SB cartridges. Column: 8x250 mm OSTION LG KS 0800 Ca2+ form Eluent: water Temperature: 80° C Flow rate: 1 mL/min. Detection: RI Peaks: 1) Saccharose, 2) Glucose, 3) Fructose, 4) Ethanol, 5) Unidentified

Fig. 23. Separation of Sugars in Wine. Direct injection of untreated sample. Column: 8x250 OSTION LG KS 0800 Ca2+ form Eluent: water Temperature: 80°C Flow rate: 1 mL/min. Detection: RI

Fig. 24. Separation of Organic Acids in Wine. Sample filtered through HEMA-cart DEAE and eluted with 0.2 M hydrochloric acid. Column: 8x250 OSTION LG KS 0800 H+ form Eluent: 10 mM sulfuric acid Temperature: 80°C Flow rate: 1 mL/min. Detection: UV at 210 nm

Fig. 25. Separation of Organic Acids in Wine. Direct injection of untreated sample. Column: 8x250 OSTION LG KS 0800 H+ form Eluent: 10 mM sulfuric acid Temperature: 80° Flow rate: 1 mL/min. Detection: UV at 210 nm

Saccharides, Organic Acids

18

ALTERNATIVE METHODS

Saccharides The octadecyl bonded reversed phase columns are often employed for separations of oligosaccharides. The high carbon content octadecyl reversed phase TESSEK Separon SGX RPS is especially suitable for this purpose. Monosaccharides can be separated on amino bonded phases in acetonitrile-water eluent. TESSEK offers TESSEK Separon SGX NH2 for this analysis. Separation of saccharides as borate complexes can be achieved on anion exchangers. Both TESSEK Separon HEMA-S 1000 DEAE and TESSEK Separon HEMA-S 1000 Q hydroxyethyl methacrylate-based anion exchangers are suitable for this purpose.

Organic acids The most common method for organic acid separation is the reversed phase chromatography on octadecyl bonded phases. Low pH low organic content eluents are used. The high carbon content TESSEK Separon SGX RPS offers higher retention times due a stronger hydrophobicity. Anion exchange chromatography is another possibility. Any anion exchanger with a suitable eluent (usually salt like sulfate or perchlorate at neutral pH) can be applied for this purpose. Both TESSEK Separon HEMA BIO 1000 DEAE and TESSEK Separon HEMA BIO 1000 Q can be successfully applied.

GUARD COLUMNS TESSEK offers two formats of stainless steel guard columns packed with both TESSEK Separon hydroxyethyl methacrylate HEMA and TESSEK Separon silica SGX sorbents for analytical purposes. The modern design ensures low dead volumes and compatibility with practically all types of chromatographic instruments.

Guard columns are supplied in a plastic box: two pieces with connecting elements and with test report. Two sizes of 4 mm ID guard columns are available with 40 mm or 50 mm length. Both sizes can be repacked on request for more economical use of stainless steel guard columns vs. CGC precolumns.

4x40 mm Stainless Steel Guard Column

TESSEK Separon Guard Column Particle size 5 µ 7 µ 10 µ SGX 161 300003 161 300004 161 300005 SGX C18 161 300013 161 300014 161 300015 SGX CN 161 300023 161 300024 161 300025 SGX NH2 161 300033 161 300034 161 300035 SGX RPS 161 300043 161 300044 161 300045 SGX CX 161 300053 161 300054 161 300055 SGX AX 161 300063 161 300064 161 300065 SGX Phenyl 161 300073 161 300074 161 300075 SGX C8 161 300083 161 300084 161 300085

4x50 mm Stainless Steel Guard Column

TESSEK Separon Guard Column Particle size 5 µ 7 µ 10 µ SGX 151 300003 151 300004 151 300005 SGX C18 151 300013 151 300014 151 300015 SGX CN 151 300023 151 300024 151 300025 SGX NH2 151 300033 151 300034 151 300035 SGX RPS 151 300043 151 300044 151 300045 SGX CX 151 300053 151 300054 151 300055 SGX AX 151 300063 151 300064 151 300065 SGX Phenyl 151 300074 151 300075 SGX C8

151 300073 151 300083 151 300084 151 300085


19

SPHERICAL SILICA SORBENTS Silica based sorbents are still the most used packing materials in liquid chromatography today. They offer excellent efficiency and mechanical stability. TESSEK Separon SGX sorbents are based on a totally porous, spherical silica. The sorbent is prepared by original technology, ensuring high batch-to-batch reproducibility. It features high sorption capacity, good mechanical strength and chemical stability, wide pore size distribution. The average pore diameter is around 8 nm, the average surface area 500 m2 and the pore volume 1.0 mL/g. Standard particle sizes are 5, 7, 10 and 60 µm; others are available for specific purposes. Normal Phase Chromatography TESSEK Separon SGX is unmodified silica for adsorption chromatography of low molecular weight solutes in a normal phase mode. Typical applications include separation of aromatic and polyaromatic hydrocarbons, lipids, steroids, fat soluble vitamins, glycoside derivatives, drugs and pharmaceuticals. TESSEK Separon SGX CN and SGX NH2 are two medium polar-bonded phases for separation of low molecular weight solutes in normal and reversed phase modes. The bonded phases offer higher reproducibility of retention volumes and shorter equilibration times compared to unmodified silica. TESSEK Separon SGX CN contains cyanoethyl groups and TESSEK Separon SGX NH2 bonded aminopropyl groups. Typical applications include separations of aromatic hydrocarbons and their derivatives, pesticides, organophosphates, fat soluble vitamins, drugs, and pharmaceuticals. The TESSEK Separon SGX NH2 is also suited for separation of mono and disaccharides in acetonitrile-water eluents.

Fig. 26. Separation of Tocopherol Isomers in Vegetable Oils. CGC Column 1x150 mm SGX 5 µm Eluent: heptane with 0.5% isopropanol Flow rate: 50 µL/min Detection: fluorometric Peaks: 1) alpha-tocopherol, 2) gamma-tocopherol, 3) delta-tocopherol For details see Application Note No 42.

Fig. 27. Determination of Sugars and Molasses. CGC Column 3x150 SGX NH2 5 mm Eluent: acetonitrile-water-80x20 Flow rate: 0.5 mL/min. Detection: RI Peaks: Fig. A: 1) solvent, 2) fructose, 3) glucose, 4) saccharose, 5) maltose, 6) lactose Fig. B: 1) solvent, 2) fructose, 3) sacccharose

Spherical Silica Sorbents – Normal Phase

20

Specifications and Ordering information

Sorbent Functional group Typical carbon content Functional group content SGX CN cyanoethyl 9% 1.0-1.3 mmol/g SGX NH2 aminopropyl 6.5% 1.5-2.2 mmol/g

Bulk sorbents - TESSEK Separon SGX is available in 10 g, 100 g, and bulk quantities.

TESSEK Separon 5 µm 7 µm 10 µm 60 µm SGX 300003 300004 300005 310003 SGX CN 300023 300024 300025 310023 SGX NH2 300033 300034 300035 310033

Packed columns Steel columns are packed individually, each with test report and chromatogram. CGC system glass columns are packed individually with test chromatogram. Preparative columns with 25 or 46 mm internal diameter are available on request. The sorbents in 60 µm particle size are available in Silica-cart cartridges for SPE. See Page 29 for Plastic Cartridges.

TESSEK Separon SGX 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300004 STEEL column 4x250 mm 141 300003 141 300004 141 300005 CGC column 3x150 mm 901 300003 901 300004 901 300005 STEEL column 4x40 mm 161 300003 161 300004 161 300005 CGC column 1x150 mm 941 300003 CGC column 1x30 mm 951 300003

TESSEK Separon SGX CN 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300024 STEEL column 4x250 mm 141 300023 141 300024 141 300025 CGC column 3x150 mm 901 300023 901 300024 901 300025 STEEL column 4x40 mm 161 300023 161 300024 161 300025 CGC column 1x150 mm 941 300023 CGC column 1x30 mm 951 300023

TESSEK Separon SGX NH2 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300034 STEEL column 4x250 mm 141 300033 141 300034 141 300035 CGC column 3x150 mm 901 300033 901 300034 901 300035 STEEL column 4x40 mm 161 300033 161 300034 161 300035 CGC column 1x150 mm 941 300033 CGC column 1x30 mm 951 300033

Spherical Silica Sorbents – Reversed Phase

21

Reversed Phase Chromatography TESSEK Separon SGX C8, SGX C18, SGX RPS, SGX Phenyl -are reversed phase sorbents differing in functionality and carbon loading. TESSEK Separon SGX C18 is octadecyl modified, the most commonly used sorbent with the widest range of applications, including pesticides, amino acid derivatives, peptides, catecholamines, drugs, pharmaceuticals, steroids, antibiotics, organic acids, nucleotides, and many others. For separation of basic compounds, the use of mobile phase modifiers (e.g., triethylamine) or low pH eluents is recommended.

Fig. 28. Separation of Fatty Acids in Blood Vessel Wall by HPLC as Phenacyl derivates. Steel Column 4x250 mm+ 4x50 mm SGX C18, 5 µm Flow rate:1.0 mL/min. Eluent: methanol-acetonitrile water. Detection: UV at 242 nm Injection: 10 µL C6 – C22 saturated and unsaturated fatty acids. For details, see Application Note No. 11. TESSEK Separon SGX RPS is octadecyl modified with very high carbon content. It offers higher retention in comparison to SGX C18 and is especially suited for ion pair chromatography and separation of oligosaccharides and other polar compounds in highly aqueous eluents. Due to its polymeric character it offers good selectivity for polyaromatic hydrocarbons. TESSEK Separon SGX C8 is an octyl modified sorbent, which offers lower retention and changed selectivity, advantageous for separation of catecholamines, peptides and some pharmaceuticals. TESSEK Separon SGX Phenyl with bonded phenyl groups offers lower retention and changed selectivity, useful for polycondensed aromatic hydrocarbons, polar aromatic compounds, and unsaturated fatty acids.

Fig. 29. Separation of Maltodextrins Steel Column 4x250 mm SGX RPS 5 µm Eluent: water Flow rate: 0.5 mL/min. Detection: RI Sample: maltodextrins, MT3–MT7 Injection: 10 µL

Fig. 30. Separation of Flunitrazepam, Diazepam, and Metabolites in Serum or Plasma. CGC Column 3x150 mm + Steel Column 4x40 SGX C18, 7µm Eluent: Methanol-NaH2PO4, pH 7.0 Detection: Diode Array at 220nm Injection: 150 µL of extract. For details, see Application Note No. 44.

Spherical Silica Sorbents – Reversed Phase

22

Specifications

Sorbent Functional group Typical carbon content SGX C18 SGX RPS SGX Phenyl SGX C8

octadecyl octadecyl

phenyl octyl

18% 24% 15% 12%

Ordering information - Bulk sorbents TESSEK Separon SGX is available in 10 g, 100 g, and bulk quantities.

TESSEK Separon 5 µm 7 µm 10 µm 60 µm SGX C18 300013 300014 300015 310013 SGX RPS 300043 300044 300045 310043 SGX Phenyl 300073 300074 300075 310073 SGX C8 300083 300084 300085 310083

Ordering information - Packed columns Steel columns are packed individually, each with test report and chromatogram. CGC system glass columns are packed individually with test chromatogram. Preparative columns with 25 or 46 mm internal diameter are available on request. The sorbents in 60 µm particle size are available in Silica-cart cartridges for SPE. See Page 29 for Plastic Cartridges.

TESSEK Separon SGX C18 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300014 STEEL column 4x250 mm 141 300013 141 300014 141 300015 CGC column 3x150 mm 901 300013 901 300014 901 300015 STEEL column 4x40 mm 161 300013 161 300014 161 300015 CGC column 1x150 mm 941 300013 CGC column 1x30 mm 951 300013

TESSEK Separon SGX RPS 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300044 STEEL column 4x250 mm 141 300043 141 300044 141 300045 CGC column 3x150 mm 901 300043 901 300044 901 300045 STEEL column 4x40 mm 161 300043 161 300044 161 300045 CGC column 1x150 mm 941 300043 CGC column 1x30 mm 951 300043

TESSEK Separon SGX Phenyl 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300074 STEEL column 4x250 mm 141 300073 141 300074 141 300075 CGC column 3x150 mm 901 300073 901 300074 901 300075 STEEL column 4x40 mm 161 300073 161 300074 161 300075 CGC column 1x150 mm 941 300073 CGC column 1x30 mm 951 300073


23

TESSEK Separon SGX C8 5 µm 7 µm 10 µm STEEL column 8x250 mm 101 300084 STEEL column 4x250 mm 141 300083 141 300084 141 300085 CGC column 1x150 mm 901 300083 901 300085 STEEL column 4x40 mm 161 300083 161 300085 CGC column 1x150 mm 941 300083 CGC column 1x30 951 300083

ION EXCHANGE CHROMATOGRAPHY TESSEK Separon SGX CX is a strongly acidic cation exchanger with a benzenesulfonic acid functional group. TESSEK Separon SGX AX is a strongly basic anion exchanger with aliphatic quaternary amine functional group. These sorbents were specially evaluated for ion exchange chromatography of ionic compounds both in aqueous and organic eluents.

Specifications and ordering information

TESSEK Separon Functional group Typical carbon content Functional group content

SGX CX SGX AX

benzensulfo quat amine

10% 8%

0.4 - 0.8 mmol/g 0.4 - 0.7 mmol/g

Bulk sorbents: TESSEK Separon SGX are available in 10 g, 100 g, and bulk quantities.

TESSEK Separon 5 µm 7 µm 10 µm 60 µm

SGX CX SGX AX

300053 300063

300054 300064

300055 300065

310053 310063

Packed columns

Steel columns are packed individually, each with test report and chromatogram. CGC system glass columns are packed individually with test chromatogram. Preparative columns with 25 or 46 mm internal diameter are available on request. The sorbents in 60 µm particle size are available in Silica-cart cartridges for SPE. See Page 29 for Plastic Cartridges.

TESSEK Separon SGX CX 5 µm 7 µm 10 µm

STEEL column 8x250 mm 101 300054 STEEL column 4x250 mm 141 300053 141 300054 141 300055 CGC column 3x150 mm 901 300053 901 300054 901 300055 STEEL column 4x40 mm 161 300053 161 300054 161 300055 CGC column 1x150 mm 941 300053 CGC column 1x30 mm 951 300053

TESSEK Separon SGX AX 5 µm 7 µm 10 µm

STEEL column 8x250 mm 101 300064 STEEL column 4x250 mm 141 300063 141 300064 141 300065 CGC column 3x150 mm 901 300063 901 300064 901 300065 STEEL column 4x40 mm 161 300063 161 300064 161 300065 CGC column 1x150 mm 941 300063 CGC column 1x30 mm 951 300063

Spherical Silica Sorbents - Macroporous

24

MACROPOROUS SILICA SORBENTS

Elution volume, mL

Fig. 31. Calibration Curves for Polystyrenes in Tetrahydrofuran. Steel Column 4x250 mm SGX 7 µm; SGX 200, 7 µm; SGX 500, 7 µm; SGX 1000, 7 µm Flow rate: 0.5 mL/min. Detection: UV at 254 nm. Based on the TESSEK Separon SGX, these sorbents are prepared by special treatment to provide large-pore material with a narrow pore size distribution. TESSEK Separon SGX 200 is prepared by hydrothermal treatment and is intended for size exclusion chromatography of organic solvent soluble polymers in the molecular weight range 1 - 100 kDa.

Fig. 32. Separation of Polystyrene Standards. Steel Column 4x250 mm, SGX 500, 7 µm Eluent: tetrahydrofuran Flow rate: 0.5 mL/min. Detection: UV at 254 nm Injection: 10 µL of standard mixture Peaks: 1) PS 1 090 000, 2) PS 173 000, 3) PS 28 000, 4) PS 9 000, 5) PS 3 770, 6) toluene. TESSEK Separon SGX 500 and 1000 are two macroporous silicas for size exclusion chromatography of organic solvent soluble polymers in the molecular weight ranges 10 - 1000 kDa and 50 - 3 000 kDa respectively

Specifications and ordering information - Bulk Sorbents

TESSEK Separon SGX are-available in 10 g, 100 g, and bulk quantities.

TESSEK Separon Pore size Pore volume Surface area

SGX 200 20 nm 0.8 mL/g 300m2/g SGX 500 50 nm 0.4 mL/g 150m2/g SGX 1000 100 nm 0.4 mL/g 150m2/g

TESSEK Separon 5 µm 7 µm 10 µm

SGX 200 320003 320004 320005 SGX 500 340003 340004 340005 SGX 1000 360003 360004 360005

Packed columns Steel columns are packed individually, each with test report and chromatogram. CGC system glass columns are packed individually with test chromatogram.

Preparative columns with 25 or 46 mm internal diameter are available on request. The sorbents in 60 µm particle size are available in Silica-cart cartridges for SPE. See Page 29 for Plastic Cartridges.

TESSEK Separon SGX 200, 7 µm SGX 500, 7 µm SGX 1000, 10 µm STEEL column 8x250 mm 101 320004 101 340004 101 360004 STEEL column 4x250 mm 141 320004 141 340004 141 360004

Polymeric Sorbents for Gas Chromatography

25

POLYMERIC SORBENTS FOR GAS CHROMATOGRAPHY The TESSEK Separon polymeric sorbents for gas chromatography are based on macroporous, spherical particles, differing widely in polarity and surface area. Generally they are applicable for separations of low boiling point liquids and gases. The temperature stability is to 200° C. No bleeding of stationary phase occurs within the working temperature range.

Fig. 33. Determination of Methanol in Spirits by GC-FID. Glass Column 4x1200 mm, TESSEK Separon SDA 100 µm, Carrier Gas: nitrogen Flow rate: 40 mL/min, Detection: FID, Temperatures: Injector 150° C, Column 130° C, Detector 180° C Injection: 0.2 µL Fig. A: Aqueous Solution of 1) 0.5% methanol, 2) 40% ethanol. Fig. B: Plum Brandy (Slivovitz), Peaks: see Fig A. For details see Application Note 36. Six types are supplied as a bulk with particles sizes 100, 150, and 200 µm. TESSEK Separon BD - co-polymer of butylmethacrylate and divinyl benzene. Nonpolar sorbent with surface area of about 60-120 m2/g. TESSEK Separon AE - co-polymer of acrylonitrile and ethylene dimethacrylate. Polar sorbent suitable for volatile liquids and gases. TESSEK Separon SE - copolymer of styrene and ethyleneglycol dimethacrylate. Medium polar sorbent also suitable for reversed phase liquid chromatography.

TESSEK Separon AD - co-polymer of acrylonitrile and divinylbenzene. Semi polar sorbent with high surface area around 400 m2/g. TESSEK Separon CHN - co-polymer of acrylonitrile and divinylbenzene especially suitable for analyses of gases and elemental (CHN) analyses. The sorbent has surface area around 650 m2/g. TESSEK Separon SDA - ternary co-polymer of styrene, divinylbenzene and acrylonitrile, especially suitable for analyses of hydrocarbons and their derivatives. The sorbent has surface area 80-90 m2/g.

Ordering information – Bulk Sorbents

The following sorbents are available in 10 g, 100 g, and bulk quantities.

TESSEK Separon Particle size µm Cat. NumberBD 100 511004

150 511005 200 511006

AE 100 512004 150 512005 200 512006

SE 100 513004 150 513005 200 513006

AD 100 514004 150 514005 200 514006

CHN 100 515004 150 515005 200 515006

SDA 100 516004 150 516005 200 516006

Packed Columns and CGC Column Holders

26

PACKED COLUMNS Stainless Steel Analytical And Semipreparative Columns

TESSEK offers two formats of stainless steel columns packed with TESSEK Separon SGX silica sorbents for analytical and semi-preparative purposes. The modern design ensures low dead volumes and compatibility with principally all types of chromatographic instruments. The exchangeable coupling fittings enable coupling with 1.5 mm or 1/16" OD capillaries. Either M5 or UNF 10-32 male thread bushings can be used. The columns are delivered single-packed with an individual test report and equipped with the UNF 10-32 coupling fittings. (Please specify this when ordering M5 coupling fittings). The 4 mm ID x 250 mm column format is still the most commonly used for analytical purposes. The 4 mm ID x 40 mm is used as guard column size. The 8 mm ID x 250 mm columns are especially suited for semi-preparative separations and size exclusion chromatography. The 8 mm ID x 80 mm column format is well suited for work with polymer based sorbents for analytical and semipreparative purposes with gradient elution. Stainless Steel Preparative Columns Besides analytical applications, HPLC chromatography is an indispensable tool also for preparation of small quantities of purified compounds. For laboratory scale purifications, preparative columns enable purification of 10-100 mg of compounds in one run. The columns are manufactured from 316 grade stainless steel, which guarantees excellent chemical inertness and corrosion stability. The columns are available packed with the complete range of TESSEK Separon SGX or TESSEK Separon HEMA sorbents. In 25x250 mm format, fixed or hand-tight end fittings are available. In the 46x300 mm format, only hand-tight fittings are available. Other column lengths are available on request.

Stainless Steel Column Packed with Sorbent

Cat. Number of Column +Cat. Number

of Packed Sorbent

8x250 mm 8x500 mm

8x80 mm 4x250 mm 4x150 mm 4x100 mm

4x50 mm 4x40 mm

101+ 102+ 109+ 141+ 143+ 149+ 151+ 161+

Inert PEEK Analytical and Semipreparative Columns

Biocompatible columns packed with TESSEK Separon HEMA Sorbents. All column parts are metal-free and made of PEEK (poly-ether-ether-ketone). The column will stand max 90 bar continuously and is excellently suited for all common HPLC and LC solvents. However, use of concentrated nitric and sulfuric acids, tetrahydrofuran and dime-thylsulfoxide is not recommended. The column dimensions include 4.6 mm ID and 30, 50, 100, 150, and 250 mm lengths, as well as 7.5 mm ID and 50, 75, 250, and 300 mm lengths. Inert Titanium Analytical And Semipreparative Columns

In special cases, stainless steel is not inert enough. TESSEK offers Titanium columns for applications requiring a higher degree of chemical resistance than available with stainless steel. When aggressive solvents are used or metal sensitive proteins are to be separated, inert materials must be used thorough all the chromatographic systems. The Titanium Zirconium alloy offers high inertness combined with high mechanical strength. The 8x250 mm columns are well suited for semipreparative and size exclusion separations, while the 8x80 mm format is ideal for gradient elutions. All column parts are machined from titanium-zirconium alloy. Ceramic paper, PTFE sieves, and PTFE sealing rings are used to support the sorbent. The column material is resistant to most eluents, including halide or low pH organic acids. The column may be attacked by hydrogenfluoric acid.

Titanium-Zirconium Column Packed with Sorbent

Cat. Number of Column + Cat. Number

of Packed Sorbent

8x250 mm 8x80 mm

201+ 209+


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COMPACT GLASS CARTRIDGE SYSTEM The TESSEK-patented design of compact glass cartridge (CGC) offers the unique combination of a low-cost and high-performance HPLC column. The CGC columns exhibit high pressure resistance (up to 30 Mpa [4400 psi]), total inertness, chemical resistance, high efficiency and, due to the smaller diameter, increased sensitivity. Moreover, any damage to the column bed is instantly visible. The columns are closed with fine mesh stainless steel sieves (silica-based sorbents). Three types of CGC cartridges packed with TESSEK Separon SGX and HEMA sorbents are available: 3x150 mm Standard analytical column 3x70 mm Column for fast separations

(custom made only) 3x30 mm Guard column or for fast analyses

of simple mixtures

Compact Glass Column Packed with Sorbent


of Packed Sorbent

3x150 mm 3x70 mm 3x30 mm

901+ 909+ 911+

Microbore columns offer greater mass sensitivity and low solvent consumption than standard analytical columns. They are indispensable when sample size is limited, expensive, or rare, or when highly hazardous solvents are used. Best results are obtained with specially constructed micro chromatographs. However, standard equipment can be used if low volume injectors and detector cells (below 1 µL) are used and dead volumes are eliminated. The pump must be able to deliver solvent at flow rates from 10 µL/min.

Compact Glass Column Packed with Sorbent


of Packed Sorbent

1x150 mm 1x30 mm

941+ 951+

CGC COLUMN HOLDERS

Fig. 34. CGC Column Holder for Combination of Guard Column 30mm + Analytical Column 150 mm UNF. Parts: 1) coupling fitting UNF 10-32, 2) screw cap, 3) holder body 30 mm, 4) holder body connector, 5) coupling insert, 7) holder body 150 mm For operating the CGC cartridges, an appropriate column holder must be used. New column holder sets are completely machined from stainless steel. For coupling to the instrument, 1.5 mm or 1/16" OD capillaries can be used. These sets are supplied in two variants: M5 Coupling fittings are supplied with M5 inner

thread. Each set contains 3 pieces of bushing M5, stainless steel and PTFE ferrules

UNF Coupling fittings are supplied with UNF 10-32 inner thread.

The following sets are available:

CGC column holder 30 mm CGC column holder 70 mm CGC column holder 150 mm

These sets are intended for the operation of coupled CGC columns, especially for CGC 3x30 guard column coupling. They can also be used for the operation of a single column. In combination with another column holder body, they can be used with any combination of CGC columns. Each of these sets contains two holder bodies of specified lengths, two screw caps, two column fittings, one column coupling union, and one holder body connector.


28


CGC Column Holder Cat. NumberCGC column holder 30 mm M5 900 001011 CGC column holder 30 mm UNF 900 001021 CGC column holder 70 mm M5 900 001013 CGC column holder 70 mm UNF 900 001023 CGC column holder 150 mm M5 900 001017 CGC column holder 150 mm UNF 900 001027 CGC column holder 70+30 mm M5 900 001015 CGC column holder 70+30 mm UNF 900 001025 CGC column holder 150+30 mm M5 900 001019 CGC column holder 150+30 mm UNF 900 001029

CGC Guard Columns For Stainless Steel. Ti-Zr And Peek Columns Disposable CGC system glass guard columns are available for use with TESSEK Separon HEMA-packed stainless steel columns. Guard columns packed with TESSEK Separon HEMA BIO 1000 are intended as universal for all SEC, IEC, HIC, IMAC and affinity

chromatography columns. TESSEK Separon HEMA BIO 1000 SB and Q ion exchanger packed guard columns are used when ionic impurities are to be trapped (IC, IEX). For use with 4 - 4.6 mm ID columns, 10 µm particle size is recommended, while for 7.5 - 8 mm ID columns 20 µm is intended for preconcentration in anion chromatography, while TESSEK Separon HEMA BIO 1000 SB 60 µm is intended as an ammonia trap in sample preconcentration for anion chromatography (see Application Note No. 57). For TESSEK stainless steel columns direct coupling column holders are available for one or two coupled guard cartridges. First set contains one 30 mm holder body, two screw caps, one coupling fitting (UNF 10-32 thread) and one column coupling union, the second contains two 30 mm holder bodies, two screw caps, one coupling fitting (UNF 10-32 thread), one column coupling insert, one holder body connector and one column coupling union. Standard CGC column holder 30 mm can be used with a short capillary connection for other manufacturers’ columns.

CGC Guard Column Holder Cat. Number

CGC Guard column holder 30 mm CGC Guard column holder 30 +.30 mm

900 001022 900 001026

CGC 3x30 mm Guard TESSEK Separon Particle Size Cat. Number

HEMA BIO 1000 10 µm 911 162005 HEMA BIO 1000 20 µm 911 162008 HEMA BIO 1000 SB 10 µm 911 162135 HEMA BIO 1000 SB 20 µm 911 162138 HEMA BIO 1000 SB 60 µm 911 172133 HEMA BIO 1000 Q 10 µm 911 162095 HEMA BIO 1000 Q 20 µm 911 162098 HEMA-S 1000 Q-L 20 µm 911 161108

Guard columns are packed immersed in 0.01% sodium azide and sealed in aluminum foil bags as a set of 3.

HEMA-cart packed with Sorbent TESSEK Separon Color Code Cat. Number

HEMA BIO 1000 DEAE blue-blue 421 172043HEMA BIO 1000 CM green-green 421 172053 HEMA BIO 1000 Q blue-black 421 172093 HEMA BIO 1000 SB green-black 421 172133 HEMA BIO 1000 E grey-grey 421 172013 HEMA BIO 1000 E-L black-black 421 172023 HEMA BIO 1000 E-H black-red 421 172083 HEMA BIO 1000 VS grey-red 421 172113 HEMA 1000 C18 orange-brown 421 170153

The SILICA-cart system consists of a prepacked 1 mL polypropylene cartridge (medical grade Tatren PD 140) packed with 60 µm spherical silica TESSEK Separon SGX sorbents.

Plastic Cartridges

29

PLASTIC CARTRIDGES

Fig. 35. Cutaway of Silica-cart and HEMA-cart The SILICA-cart and HEMA-cart system is specially designed to optimize sample preparation and storage: The cartridge works by extracting certain classes of compounds from injected samples while allowing others to pass through. The system can be used to concentrate trace components from environmental and biological samples. It is ideal for the preparation of samples for analysis by HPLC, GC, TLC, RIA and other methods. The cartridge is terminated with a male and female standard syringe cone (Luer) and can be used with a syringe, a peristaltic

Pump, or a suitable vacuum-operated manifold. The maximum operating pressure is 0.3 Mpa (43 psi); the suction mode operation is recommended. The cartridges are supplied with tight caps to ensure long-term sample storage.

Fig. 36. Separation of Complex Protein Mixture on HEMA-cart SB. Sample: 1) ovalbumin, 2) hemoglobin, 3) cytochrome C, 4) lysozyme Load volume: 400 µL Flow: 1 mL/min., other conditions see HEMA-cart leaflet. The HEMA-cart system consists of a pre-packed 1 mL polypropylene cartridge (medical grade Tatren PD 140) packed with 60 µm TESSEK Separon HEMA-BIO 1000 modified sorbents.

HEMA-cart packed with TESSEK Separon Color Code Cat. Number

HEMA BIO 1000 DEAE blue-blue 421 172043 HEMA BIO 1000 CM green-green 421 172053 HEMA BIO 1000 Q blue-black 421 172093 HEMA BIO 1000 SB green-black 421 172133 HEMA BIO 1000 E grey-grey 421 172013 HEMA BIO 1000 E-L black-black 421 172023 HEMA BIO 1000 E-H black-red 421 172083 HEMA BIO 1000 VS grey-red 421 172113 HEMA 1000 C18 orange-brown 421 170153

The SILICA-cart system consists of a prepacked 1 mL polypropylene cartridge (medical grade Tatren PD 140) packed with 60 µm spherical silica TESSEK Separon SGX sorbents.

SILICA-cart packed with TESSEK Separon Color Code Cat. Number

SGX yellow-yellow 421 310003 SGX C18 orange-orange 421 310013 SGX CN yellow-orange 421 310023 SGX NH2 yellow-brown 421 310033 SGX RPS orange-black 421 310043 SGX C8 yellow-black 421 310083 SGX CX green-orange 421 310053 SGX AX blue-orange 421 310063 SGX Phenyl orange-grey 421 310073

Both HEMA and silica-cart cartridges are supplied in boxes of 6 or in polyethylene bags of 100.

Application Notes – Alphabetical by Compound

30

The following table is an alphabetical list of compounds. To obtain a copy of any of these application notes, send an e-mail to [email protected] indicating the number.

Analyte Sample No. Product

Abscisis acid plant extracts 68 silica-cart SGX C18, 3x150 SGX C18, 5 µm

Acrylate steroidal 17betametyl det. Z and E isomers 103 4x250 SGX C18, 10 µm Aflatoxin Ml milk 9 silica-cart SGX C18 Aflatoxins (very low concentrations) in suckling's nutrition 8 3x150 SGX 5 µm Alkaloids(opium) poppy straw 60 3x150 SGX C18, 5 µm Alkaloids-basic colchicinoid plant extracts 129 4x250 SGX C18,10 µm Alkaloids-neutral colchicinoid plant extracts 128 4x250 SGX C18, 7 µm Alpha-tocopherol, retinol human serum 25 3x150 SGX 5 µm Alpha-tocopherol, retinol, beta-carotene human serum 41 3x150 SGX C18 7 µm Anisole p-bromoanisole after direct brominat. 81 4x250 SGX C18, 5 µm Antiepileptics serum 72 3x150 SGX C18, 7 µm Antiepileptics and Barbiturates plasma or serum 71 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm Ascorbic acid foodstuffs 84 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm Aspartame, saccharine foodstaffs 91 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm ATP derivatives separation 18 3x150 SGX C18, 7 µm Barbiturates and some antiepileptics plasma or serum 71 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm Benzo(a)pyrene liquid smoking preparation DTP-1 14 3x150 SGX C18, 5 µm Benzo(a)pyrene smoked meat products 16 3x150 SGX C18, 5 µm Benzoic and sorbic acids foodstuffs 54 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm Benzoquinone, dihydroxybenzenes photochemical reactions of phenol 97 3x150 SGX C18, 7 µm Beta-1,3-glucans (water soluble) gel permeation chromatography 56 8x250 HEMA-S 300,1000,10 µm Beta-carotene, retinol, alpha-tocopherol human serum 41 3x150 SGX C18, 7 µm Betamethasone Dipropionate ointments 79 3x150 SGX C18, 5 µm Bile acids (free and conjugated) physiological fluids 4 silica-cart SGX C18 Biphenyl (degradation products) separation 80 4x250 SGX C18,10 µm Bitter acids(alpha and beta) hops 93 3x150 SGX C18, 5 µm;

3x30 SGX C18, 5 µm Caffeine in urine 88 3x150 SGX C18, 7 µm Caffeine foodstuffs 53 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7 µm Carbisocaine, heptacaine, pentacaine biological fluids 45 silica-cart SGX C18 Carbofurane soil and hop leaves 131 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7 µm Catecholamines (free plasma via condensation

with 1,2-diphenylen diamine 95 3x150 SGX C18, 5 µm

Catecholamines, tyrosine metabolites urine 73 1x150 SGX C18, 5 µm Cationactive surfactants 113 silica-cart SGX C18 Cellulose lithium chloride-dimethyl acetamide el 62 8x250 HEMA 81040,

8x250 HEMA 8101000, 10 µm Cellulose cadoxen 63 4x250 HEMA 40, 100, HEMA-S 300,

HEMA-S 1000,10 µm Cellulose(sodium Carboxymethyl) SEC characterization 61 8x250 HEMA BIO100, 300, 1000,

10 µm Cocaine and benzoylecnogine biological fluids 110 3x150 SGX CN, 7 µm,

3x30 SGX CN, 7 µm Cortisol serum 124 3x150 SGX C18, 7 µm Cotinine human urine 118 3x150 SGX C18, 5 µm Creatinine serum 109 3x150 HEMA BIO1000 SB,10 µm


31

Analyte Sample No. Product Cyanocobalamine fermented vegetable drinks 116 3x150 SGX, 7 µm;

3x150 SGX C18, 5 µm Cyclosporine A blood 132 4x250 SGX C18, 5 µm Cyclosporines separation 104 3x150 SGX C8,

3x150 SGX C18, 5 µm Cyclosporins fermentation broth of typocladium infl 90 3x150 SGX C18, 7 µm Cytokinins plant cell 38 4x250 SGX C18, 7 µm D/L ratio of select.dansyl amino acids dentin 120 3x150 SGX RPS, 5 µm Dansyl derivatives of amino acides dentin from antiant teeth 119 3x150 SGX RPS, 5 µm Dexamethasone acetate dexameth. pix ointment 77 silica-cart SGX C18,

3x150 SGX C18, 5 µm Dexamethasone acetate, triamcinolon acetonide ointments 78 3x150 SGX C18, 5 µm Diazepam, flunitrazepam, metabolites plasma or serum 44 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7pm Digoxin injection solutions 85 3x150 SGX C18, 7 µm Dihydroxybenzenes, benzoquinone photochemical reactions of phenol 97 3x150 SGX C18, 7 µm Dioxacarb aqueous samples 51 silica-cart SGX C18 Dipeptides(selected group) separation 29 8x80 and 8x250 HEMA BlO 1000 CM,

7pm diuretics urine 122 3x150 SGX C18, 7pm DNS-amino acids derivatives sensitive analysis 19 1x150 SGX C18,.5 µm Ecdysteroides plant cell extract 111 4x250 SGX C18, 7 µm Erythromycin A N-oxides, pseudoeryt. A hemiketal

fermentation brothon 67 8x250 SGX C18, 7 µm

Ethanol and methanol enzymatic detection 32 3x150 HEMA-S 1000, 3x30 HEMA BIO1000 E, 10 µm

Ethimizol and its metabolites biological fluids 48 silica-cart SGX C18, 4x250 SGX, 5 µm

Ethylenethiourea beverages 28 8x80 HEMA 8101000 DEAE, 7pm Ethynilestradiol, levonorgestrel coated tablets Neogest 89 3x150 SGX C18, 5 µm Fatty acids blood vessel wall (as phenacyl deriv.) 11 4x250 SGX C18, 5 µm Fatty acids (medium chain) serum 59 silica-cart SGX C18 Fatty acids (very long chain) isol. from lower organisms 108 silica-cart SGX C18 Fibrinopeptides released from human fibrinogen 92 3x150 SGX C18, 7 µm Flunitrazepam, diazepam, metabolites plasma or serum 44 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7pm Gentamicin serum 47 silica-cart SGX,

4x250 SGX C18, 5 µm Glutamate sodium foodstuffs 127 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7pm Halomethasone ointments 76 3x150 SGX C18, 5 µm Hemicelluloses strongly alkaline medium 21 8x250 HEMA-S 1000, 10 µm Heptacaine, arbisocaine, pentacaine biological fluids 45 silica-cart SGX C18 Herbicides (urea) separation 87 3x150 SGX NH2 7 µm Hexobarbital and oxazepam racemic direct chiral resolution 100 3x150 SGX beta-CD, 7 µm Hippuric and methylhippuric acids urine 1 3x150 SGX C18, 5 µm Hyaluronate (sodium) fractions gel permeation chromatography 58 8x250 HEMA-S 300, 1000, 10 µm 5-Hydroxyindolacetic acid, serotonin plasma 13 3x150 SGX C18, 7 µm Chlorhexidine pharmaceutical preparations 3 3x150 SGX CN, 5 µm Chlorides and sulphates solution with malic acid 74 3x150 HEMA-S 1000 Q-L 10 µm Chlorides, nitrates and sulphates natural waters 66 3x150 HEMA-S 1000 Q-L 10 µm Indoyl—acetic acid (auxin) releated indole compounds 96 4x250 SGX C18, 7 µm Inhibitors of the enzyme cholinesterase water 52 silica-cart SGX C18 Inositolphosphates and nucleotides 32Pi labled erythrocyte membranes 106 4x250 HEMA-S 1000 Q 10 µm INPANKINE-peptide inhibitor pancreatic elastase 101 3x150 SGX C18, 5 µm Isoprenaline, isoprenalinesulphonicacid injection solutions 65 3x150 SGX C18, 7 µm Laidlomycin, 26deoxylaidlomycin determination 83 3x150 SGX C18, 5 µm Levonorgestrel, ethynilestradiol coated tablets Neogest 89 3x150 SGX C18, 5 µm


32

Analyte Sample No. Product Lignins (acetylated) SEC 31 3x150 HEMA-S 1000,10 µm Lysergic acid (simple derivatives) separation 35 3x150 SGX C18, 5 µm Mesocarb urine 121 3x150 SGX C18, 7 µm Metabolites, diazepam, flunitrazepam plasma or serum 44 3x150 SGX C18, 7 µm,

3x30 SGX C18; 7 µm Methanol spirits 36 GC-FID on glass

col. 1200x4 SDA, 100 µm Methanol and ethanol enzymatic detection 32 3x150 HEMA-S 1000,

3x30 HEMA BlO 1000 E 10 µm Methotrexate , 7-OH methotrexate serum 70 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7 µm Methylhippuric and hippuric acids urine 1 3x150 SGX C18, 5 µm 8-Methoxypsoralen human plasma 114 silica-cart SGX C18 Mevinolin and mevinolic acid fermentation liquids 107 3x150 SGX C18, 5 µm Neopterine human urine 49 silica-cart SGX C18,

4x250 SGX C18, 7 µm Nitrate residues after industr.,

military explosives 33 3x150 HEMA-S 1000 Q-L 10 µm

Nitrates hops and beer 50 3x150 HEMA-S 1000 Q-L 12 µm Nitrates, chlorides and sulphates natural waters 66 3x150 HEMA-S 1000 Q-L 10 µm Nitrogen ammoniacal fertilizers 134 3x150 HEMA BIG 1000 SB-L, 10 µm Ofloxacin serum and blood 5 3x150 SGX CN, 7 µm Oxazepam and hexobarbital racemic direct chiral resolution 100 3x150 SGX beta-CD, 7 µm PAHs fats, oils, lipid extracts 130 8x500 BI0-BEADS S-X3 Patulin from apple pulp 17 silica-cart SGX Pentacaine, carbisocaine, heptacaine biological fluids 45 silica-cart SGX C18 Pentoxifylline and its monohydroxylated metabol

serum 46 silica-cart SGX C18

Peptides HPLC at lowered temperatures 64 8x80 EDMA 40,10 µm Peptides syntetic retention time prediction 102 4x250 SGX C18, 5 µm Peptidic fragments of alpha gliadin isolation,analysis 7 4x250 SGX C18, 5 µm;

16x250 SGX C18,10 µm Phenylglyoxylic acid urine 2 3x150 HEMA-S 1000 Q-L 10 µm Phenylurea herbicides soil 98 3x150 SGX C18, 5 µm Phospholipids lung lavage fluid 27 silica-cart SGX C18 Pigments (Elderberry) separation 125 3x150 SGX C18, 7 µm Pigments (red beet) separation 126 3x150 SGX C18, 7 µm Plasmide DNA probe purification 133 8x250 HEMA BI01000, 10 µm Plasticizers industrial exhalates 117 3x150 SGX 7 µm, SGX C18, 7 µm;

silica-cart SGX C18 Polyaromatic hydrocarbons water 94 3x150 SGX RPS, 5 µm Polycyclic aromatic hydrocarbon extracts purification 15 silica-cart SGX C18 Polycyclic aromatic hydrocarbons drinking waters 115 3x150 SGX C18, 5 µm Probenecide human urine 86 3x150 SGX C18, 7 µm Proteins (wheat storage) separation 112 8x80 EDMA 2000, 7 µm Pseudoephedrine, norpseudoephedrine resolution of enantiomers 99 3x150 SGX beta-CD, 7 µm Pterins urine 40 3x150 SGX C18, 5 µm Rare earth elements deter.with post-column

derivatization 12 3x150 SGX C18, 5 µm

Retinal, alpha-tocopherol human serum 25 3x150 SGX, 5 µm Retinol, alpha-tocopherol, beta-carotene human serum 41 3x150 SGX C18, 7 µm Riboflavin human urine 24 3x150 SGX C18, 5 µm


33

Analyte Sample No. Product Saccharine, aspartame foodstuffs 91 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7 µm Serotonin, 5-hydroxyindolacetic acid plasma 13 3x150 SGX C18, 7 µm Silymarin bile 34 3x150 SGX C18, 7 µm Sorbic and benzoic acids foodstuffs 54 3x150 SGX C18, 7 µm,

3x30 SGX C18, 7 µm Steroides ointments 75 silica-cart SGX Stobadin serum 37 silica-cart SGX C18,

3x150 SGX, 5 µm Styrene oligomers analysis by adsorption

chromatogr. 20 3x150 SGX, 5 µm

Sugars plant cell viability 39 4x250 SGX NH2 7 µm Sugars molasses 43 3x150 SGX NH2 5 µm Sulphates and chlorides solution with malic acid 74 3x150 HEMA-S 1000 Q-L 10 µm Sulphates, chlorides and nitrates natural waters 66 3x150 HEMA-S 1000 Q-L 10 µm Theophyline serum and blood 23 silica-cart SGX C18 Theophylline serum 22 3x150 SGX C18, 5 µm Tocopherol isomers vegetable oils 42 1x150 SGX 5 µm Triamcinolon acetonide, dexamethasone acetate

ointments 78 3x150 SGX C18, 5 µm

Triazine pesticides analysis of a mixture 55 3x150 SGX C18, 7 µm Triazolam serum or plasma 69 3x150 SGX C18, 7 µm;

3x30 SGX C18, 7 µm Triglycerides (plant) analysis 105 4x250 SGX RPS 5 µm Tyrosine, catecholamines metabolites urine 73 1x150 SGX C18, 5 µm Urinary carboxylic acids after deriv. by 2-nitrophenyl

hydrazine 123 3x150 SGX C18, 5 µm

Vanillin lignosulphonate hydrolyzates 30 3x150 HEMA-S 1000, 10 µm Vitamin A(all trans retinol) vitamin premixes 82 3x150 SGX C18, 5 µm Vitamin D3 metabolites biological fluids 26 silica-cart SGX C18 Zearalenon (F-2toxin) wheat 10 3x150 SGX C18, 5 µm

Application Notes – Numerical Listing

34

No. 1 Gartzke J., Vins I. Determination of Hippuric and Methylhippuric Acids in Urine on CGC Column 3x150 SGX C18 5 µm

No. 2 Gartzke J., Vins I. Determination of Phenylglyoxylic Acid in Urine on CGC Column 3x150 HEMA S 1000 QL 10 µm

No. 3 Benes K., Pavelek Z. Determination of Chlorhexidine in Pharmaceutical Preparations on CGC Columns 3x150 SGX CN 5 µm

No. 4 Jirsa M., Kotal P. Isolation and Preconcentration of Free and Conjugated Bile Acids from Physiological Fluids on SILICA-cart SGX C18

No. 5 Rehak V. Determination of Ofloxacin in Serum and Blood on CGC Column 3x150 SGX CN 7 µm No. 7 Kocna P. Isolation and Analysis of the Peptidic Fragments of alpha- Gliadin Using Reversed Phase HPLC on

Steel Column 4x250 SGX C18 5 µm and Preparative Column 16x250 SGX C18 10 µm No. 8 Adensam L., Lebedova M. Determination of Very Low Concentrations of Aflatoxins in Suckling´s Nutrition on

CGC Column 3x150 SGX 5 µm No. 9 Adensam L., Lebedova M., Marko A. Isolation of Aflatoxin M1 from Milk on SILICA-cart SGX C18 No. 10 Adensam L., Lebedova M. Determination of Zearalenon(F-2 toxin) in Wheat on CGC Column 3x150 SGX C18

5 µm No. 11 Hanis T., Smrz M., Klir P., Macek K. Deyl Z. Determination of Fatty Acids in Blood Vessel Wall by HPLC as

Phenacyl Derivatives on Steel Column4x250 SGX C18 5 µm No. 12 Kuban V., Gladilovich D.B. Determination of Rare Earth Elements with Post-column Derivatization on CGC

Column 3x150 SGX C18 5 µm No. 13 Sebekova K. Determination of Serotonin and 5-Hydroxyindolacetic Acid in Plasma by HPLC on CGC Column

3x150 SGX C18 7 µm No. 14 Simko P. Isolation and Determination of Benzo(a)pyrene in Liquid Smoking Preparation UTP-1 on CGC

Column 3x150 SGX C18 5 µm No. 15 Simko P. Purification of Polycyclic Aromatic Hydrocarbon Extracts on SILICA-cart SGX C18 No. 16 Simko P. Isolation and Determination of Benzo(a)pyrene in Smoked Meat Products on CGC Column 3x150

SGX C18 5 µm No. 17 Adensam L., Lebedova M. Isolation of Patulin from Apple Pulp on SILICA-cart SGX No. 18 Soltes L. Separation of ATP Derivatives by HPLC on CGC Columns 3x150 SGX C18 7 µm No. 19 Koroleva E.M., Nesterov V.V. Sensitive Analysis of DNS-Amino Acids Derivatives by Reversed Phase HPLC on

CGC Column 1x150 SGX C18 5 µm No. 20 Nesterov V.V., Krasikov V.D. Analysis of Styrene Oligomers by Adsorption Chromatography on CGC Column

3x150 SGX 5 µm No. 21 Eremeeva T., Ebringerova A. SEC Analysis of Hemicelluloses in Strongly Alkaline Medium on Steel Column

8x250 HEMA S 1000 10 µm No. 22 Gartzke J., Vins I. Determination of Theophylline in Serum on CGC Column 3x150 SGX C18 5 µm No. 23 Gartzke J., Vins I. Isolation of Theophylline from Serum and Blood on SILICA-cart SGX C18 No. 24 Yakushina L., Bender. E. Determination of Riboflavin in Human Urine on CGC Column 3x150 SGX C18 5 µm No. 25 Yakushina L., Bender.E. Determination of alpha-Tocopherol and Retinol in Human Serum on CGC Column

3x150 SGX 5 µm No. 26 Yakushina L., Sergeev I., Terechina T., Spiritchev V. Analysis of Vitamin D3 Metabolites in Biological Fluids on

SILICA-cart SGX C18 No. 27 Sachse K., Muller G., Grajetzki C. Rapid Isolation of Phospholipids from Lung Lavage Fluid on SILICA-cart

SGX C18 No. 28 Wang H., Pacakova V., Stulik K. Direct Determination of Ethylenethiourea in Beverages on Steel Column 8x80

HEMA-BIO 1000 DEAE 7 µm No. 29 Wang H., Pacakova V., Stulik K. Separation of a Selected Group of Dipeptides on Steel Column 8x80 HEMA

BIO 1000 CM 7 µm and Steel Column 8x250 HEMA BIO 1000 CM 7 µm No. 30 Brezny R., Vodny S., Pranda J. Determination of Vanilin in Lignosulphonate Hydrolyzates on CGC Column

3x150 HEMA S 1000 10 µm No. 31 Brezny R., Pranda J., Vodny S. SEC of Acetylated Lignins on CGC Column 3x150 HEMA S 1000 10 µm No. 32 Tagliaro F., Dorizzi R., Trabetti E., Marrigo M. Enzymatic Detection of Methanol and Ethanol on Two CGC


35

Columns 3x150 HEMA S 1000 10 µm in series and CGC Column 3x30 HEMA BIO 1000 E 10 µm No. 33 Krajnik V. Nitrate Determination in Residues after Industrial and Military Explosives by Ion Chromatography on

CGC Column 3x150 HEMA S 1000 QL 10 µm No. 34 Strnadova J., Skarka P., Paulova J. Determination of Silymarin in Bile on CGC Column 3x150 SGX C18 7 µm No. 35 Flieger M. Separation of Simple Derivatives of Lysergic Acid on CGC Column 3x150 SGX C18 5 µm No. 36 Grec. V. Determination of Methanol in Spirits by GC-FID on Glass Column 1200x4 SDA 100 µm No. 37 Soltes L. Isolation and Determination of Stobadin in Serum on SILICA-cart SGX C18 and CGC Column 3x150

SGX 5 µm No. 38 Vanek T. Determination of Cytokinins on Steel Column 4x250 SGX C18 7 µm No. 39 Vanek T., Vankova R. HPLC of Sugars-Determination of Plant Cell Viability on Steel Column 4x250 SGX NH2

7 µm No. 40 Krijt J. Determination of Pterins in Urine on CGC Column 3x150 SGX C18 5 µm No. 41 Yakushina L., Kharitonchik L., Bender E. Simultaneous Determination of alpha-Tocopherol, Retinol and beta-

Carotene in Human Serum on CGC Column 3x150 SGX C18 7 µm No. 42 Yakushina L., Bender E. Direct Determination of Tocopherol Isomers in Vegetable Oils on CGC Column 1x150

SGX 5 µm No. 43 Strnadova J. Determination of Sugars in Molasses on CGC Column 3x150 SGX NH2 5 µm No. 44 Balikova M. Identification and Determination of Flunitrazepam, Diazepam and Metabolites in Serum or Plasma

on CGC Column 3x150 SGX C18 7 µm with CGC Precolumn 3x30 SGX C18 7 µm No. 45 Soltes L. Isolation of Carbisocaine ,Heptacaine or Pentacaine from Biological Fluids on SILICA-cart SGX C18 No. 46 Soltes L. Isolation of Pentoxifylline and its Monohydroxylated Metabolite from Serum on SILICA-cart SGX C18 No. 47 Soltes L. Isolation and Determination of Gentamicin from Serum on SILICA-cart SGX and Steel Column 4x250

SGX C18 5 µm No. 48 Soltes L. Isolation and Determination of Ethimizol and its Metabolites from Biological Fluids on SILICA-cart

SGX C18 and Steel Column 4x250 SGX 5 µm No. 49 Orvisky E. Determination of Neopterine in Human Urine on SILICA-cart SGX C18 and Steel Column 4x250

SGX C18 7 µm No. 50 Kroupa F. Determination of Nitrates in Hops and Beer on CGC Column 3x150 HEMA S 1000 QL 12 µm No. 51 Uchytil B. Isolation and Concentration of Dioxacarb from Aqueous Samples on SILICA-cart SGX C18 No. 52 Uchytil B. Removal of Interferences in the Determination of Inhibitors of Enzyme Cholinosterase in Water on

SILICA-cart SGX C18 No. 53 Smejkalova M. Determination of Caffeine in Foodstuffs on CGC Column 3x150 SGX C18 7 µm with Precolumn

3x30 SGX C18 7 µm No. 54 Smejkalova M. Determination of Sorbic and Benzoic Acids in Food stuffs on CGC Column 3x150 SGX C18 7

µm with Precolumn 3x30 SGX C18 7 µm No. 55 Strnadova J. Procedure for the Analysis of Triazine Pesticides on CGC Column 3x150 SGX C18 7 µm No. 56 Soltes L. GPC of Water-Soluble beta-1,3-Glucans on Steel Column 8x250 HEMA S 300 10 µm and Steel

Column 8x250 HEMA S 1000 10 µm No. 57 Pospichalova D., Vins I. Determination of Chloride at ppb Levels by IC on CGC Columns 3x150 HEMA S 1000

QL 10 µm, 3x30 HEMA S 1000 QL 25 µm,3x30 HEMA 1000 SB 60 µm No. 58 Soltes L. GPC of Sodium Hyaluronate Fractions on Steel Column 8x250 HEMA S 300 10 µm and Steel Column

8x250 HEMA S 1000 10 µm No. 59 Kmoch S. Isolation of Medium Chain Fatty Acids from Serum on SILICA-cart SGX C18 No. 60 Strnadova J. Determination of Opium Alkaloids on CGC Column 3x150 SGX C18 5 µm No. 61 Danhelka J. SEC Characterization of Sodium Carboxymetyl Cellulose on Steel Column 8x250 HEMA BIO 100

10 µm,Steel Column 8x250 HEMA BIO 300 10 µm and Steel Column 8x250 HEMA BIO 1000 10 µm in series No. 62 Danhelka J. SEC Analysis of Cellulose in Lithium Chloride-Dimethyl acetamide Eluent on Steel Column 8x250

HEMA BIO 40 10 µm and Steel Column 8x 250 HEMA BIO 1000 10 µm No. 63 Danhelka J. SEC Analysis of Cellulose in Cadoxen on Steel Column 4x250 HEMA 40 10 µm,Steel Column

4x250 HEMA 100 10 µm,Steel Column 4x250 HEMA S 300 10 µm and Steel Column 4x250 HEMA S 1000 10 µm in series


36

No. 64 Lebl M., Fang S., Hruby V.J. HPLC of Peptides at Lowered Temperatures on Steel Column 8x80 EDMA 40 10 µm

No. 65 Dolezalova M. Determination of Isoprenaline and Isoprenalinesulphonic Acid an Injection Solutions onCGC Column 3x150 C18 7 µm

No. 66 Valny Z., Vesely M., Sixta V. Determination of Chlorides,Nitrates and Sulphates in Natural Waters on CGC Column 3x150 HEMA S 1000 QL 10 µm

No. 67 Beran M., Prikrylova V., Sedmera P., Novak J., Zima J., Blumaerova M.Isolation of Erythromycin A N-Oxide and Pseudoerythromycin A Hemiketal from Fermentation Brothon on Steel Column 8x250 SGX C18 7 µm

No. 68 Rittich B., Lanickova B., Hrib J. Determination of Abscisis Acid in Plant Extracts on SILICA-cart SGX C18 and CGC Column 3x150 SGX C18 5 µm

No. 69 Balikova M. Determination of Toxic Levels of Triazolam in Serum or Plasma on CGC Column 3x150 SGX C18 7 µm with CGC Precolumn 3x30 SGX C18 7 µm

No. 70 Novak M., Neumann M. Determination of Methotrexate and 7-OH Methotrexate in Serum on CGC Column 3x150 SGX C18 7 µm with CGC Precolumn 3x30 SGX C18 7 µm

No. 71 Balikova M. Determination of Toxic Levels of Barbiturates and some Antiepileptics in Plasma or Serum on CGC Column 3x150 SGX C18 7 µm with CGC Precolumn 3x30 SGX C18 7 µm

No. 72 Novak M., Jedlickova B. Determination of Antiepileptics in Serum on CGC Column 3x150 SGX C18 7 µm No. 73 Novak M. Determination of Catecholamines and Tyrosine Metabolites in Urine by Microcolumn LC on CGC

Column 1x150 SGX C18 5 µm No. 74 Rihova J., Ondrousek S. Determination of Chlorides and Sulfates in Solution with Malic Acid on CGC Column

3x150 HEMA S 1000 QL 10 µm No. 75 Sabartova J., Vyskovska M., Kubes J. Extraction of Steroids from Ointment on SILICA-cart SGX No. 76 Sabartova J., Vyskovska M., Kubes J. Determination of Halomethasone in Ointment on CGC Column 3x150

SGX C18 5 µm No. 77 Sabartova J., Vyskovska M., Kubes J. Determination of Dexamethasone Acetate in Dexamethazon Pix

Ointment on SILICA-cart SGX C18 and CGC Column 3x150 SGX C18 5 µm No. 78 Sabartova J., Vyskovska M., Kubes J. Determination of Triamcinolone Acetonide and Dexamethasone Acetate

in Ointments on CGC Column 3x150 SGX C18 5 µm No. 79 Sabartova J., Vyskovska M., Kubes J. Determination of Betamethasone Dipropionate in Ointment on CGC

Column 3x150 SGX C18 5 µm No. 80 Tesarova E., Pacakova V., Stulik K. Separation of the Degradation Products of Biphenyl on Steel Column

4x250 SGX C18 10 µm No. 81 Drasar P. Determination of Anisole in Samples of p-Bromoanisole after the Direct Bromination on Steel Column

4x250 SGX C18 5 µm No. 82 Rittich B. Determination of Vitamin A(all trans retinol)in Vitamin Premixes by HPLC on CGC Column 3x150

SGX C18 5 µm No. 83 Beran M., Zielke R., Dornberger K., Stengel C., Zima J., Graele U.Determination of Laidlomycin and 26-

Deoxylaidlomycin by HPLC on CGC Column 3x150 SGX C18 5 µm No. 84 Smejkalova M. Determination of Ascorbic Acid in Foodstuffs on CGC Column 3x150 SGX C18 7 µm with CGC

Precolumn 3x30 SGX C18 7 µm No. 85 Dolezalova M. Determination of Digoxin in Injection Solutions on CGC Column 3x150 SGX C18 7 µm No. 86 Vodickova I., Slechtova R., Rehak V. Separation of Probenecide in Human Urine on CGC Column 3x150 SGX

C18 7 µm No. 87 Ocenaskova V., Kuzilek V. Separation of Urea Herbicides by HPLC on CGC Column 3x150 SGX NH2 7 µm No. 88 Rehak V., Vodickova I., Slechtova R. Determinace of caffeine in Urine on CGC Column 3x150 SGX C18 7 µm No. 89 Opocensky Vl., Patkova J., Jansa J. Determination of Ethinylestradiol and Levonolgestrel in Coated Tablets

Neogest on CGC Column 3x150 SGX C18 5 µm No. 90 Beran M. Determination of Cyclosporins in Fermentation Broth of Typocladium Inflatum on CGC Column 3x150

SGX C18 7 µm No. 91 Smejkalova M. Determination of Saccharine and Aspartam in Foodstuffs on CGC Column 3x150 SGX C18 7

µm with CGC Precolumn 3x30 SGX C18 7 µm No. 92 Suttnar J., Dyr J.E. Determination of Fibrinopeptides Released from Human Fibrinogen on CGC Column 3x150


37

SGX C18 7 µm No. 93 Kroupa F. Determination of Analogs of alpha and beta-Bitter Acids in Hops by Using the HPLC Method on CGC

Column 3x150 SGX C18 5µm with CGC Precolumn 3x30 SGX 18 5 µm No. 94 Kuzilek V. Separation and Determination of Polyaromatic Hydro carbons in Water on CGC Column 3x150 SGX

RPS 5 µm No. 95 Husek P. Determination of Free Plasma Catecholamines via Condensation with 1,2-Diphenylenediamine on

CGC Column 3x150 SGX C18 5 µm No. 96 Eder J., Cermak V. Determination of Indoyl-3-Acetic Acid (Auxin) and of Releated Indole Compounds on Steel

Column 4x250 SGX C18 7 µm No. 97 Brodilova J., Sedlak P. Determination of Dihydroxybenzenes and Benzoquinone in Photochemical Reactions of

Phenol on CGC Column 3x150 SGX C18 7 µm No. 98 Frantisek J., Novakova O. Determination of Phenylurea Herbicides in Soil by the HPLC Method on CGC

Column 3x150 SGX C18 5 µm No. 99 Prihoda M. Resolution of Enantiomers of Pseudoephedrine and Norpseudoephedrine on CGC Column 3x150

SGX beta- CD 7 µm No. 100 Prihoda M. Direct Chiral Resolution of Racemic Hexobarbital and Oxazepam on CGC Column 3x150 SGX

beta-CD 7 µm No. 101 Kocna P., Kasafirek E., Fric P., Slaby J. Bioavailability of INPANKIN-Peptide Inhibitor of Pancreatic Elastase on

CGC Column 3x150 SGX C18 5 µm No. 102 Kocna P., Krchnak V., Kasafirek E. Retention Time Prediction from Amino Acid Sequence in Synthetic Peptides

on Steel Column 4x250 SGX C18 5µm No. 103 Drasar P., Pouzar V. Determination of Z-and E-Isomers in Steroidal 17 beta Methyl Acrylates of St-CH=CH-

COOCH3 Type on Steel Column 4x250 SGX C18 10 µm No. 104 Flieger M., Zapletal J. Separation of Cyclosporines on CGC Column 3x150 SGX C 8 5 µm and CGC Column

3x150 SGX C18 5 µm No. 105 Rezanka T. Analysis of Plant Triglycerides by Means Reversed Phase HPLC on Steel Column 4x250 SGX

RPS 5 µm No. 106 Sevcik J., Varecka L. Separation of Inositolphosphates and Nucleotides in 32Pi labelled Erythrocyte

Membranes on Steel Column 4x250 HEMA S 1000 Q 10 µm No. 107 Kysilka R. Determination of Mevinolin and Mevinolic Acid in Fermentation Liquids on CGC Column 3x150 SGX

C18 5 µm No. 108 Rezanka T. Isolation and Preconcentration of Very Long Chain Fatty Acids from Lower Organisms on SILICA-

cart SGX C18 No. 109 Orvisky E., Bernat I. Determination of Creatinine in Serum on CGC Column 3x150 HEMA BIO 1000 SB 10 µm No. 110 Balikova M. Identification and Determination of Cocaine and Benzoylecgonine in Biological Fluids CGC Column

3x150 SGX CN 7 µm with Precolumn 3x30 SGX CN 7µm No. 111 Vanek T. Separation of Ecdysteroids in Plant Cell Extract of Pteridium Aquilinum on Steel Column 4x250 SGX

C18 7 µm No. 112 Holeckova J. Separation of Wheat Storage Proteins on Steel Column 8x80 EDMA 2000 7 µm No. 113 Koula V., Kubankova J., Kucova D., Gasparic J. Isolation and Preconcentration of Cationactive Surfactants on

SILICA-cart SGX C18 No. 114 Kucova D., Davidkova P., Koula V., Gasparic J. Liquid-Solid Extraction of 8-Methoxypsoralen from Human

Plasma before its HPLC Determination on SILICA-cart SGX C18 No. 115 Simko P. Determination of Polycyclic Aromatic Hydrocarbons in Drinking Water on CGC Column 3x150 SGX

C18 5 µm No. 116 Simko P., Drdak M., Becova K. Determination of Cyanocobalamine in Fermented Vegetable Drinks on CGC

Column 3x150 SGX 7 µm and CGC Column 3x150 SGX C18 5 µm No. 117 Fischer J., Ventura K., Jandera P., Prokes B. Determination of Plasticizers in Industrial Exhalates on SILICA-

cart SGX C18,CGC Column 3x150 SGX 7 µm and CGC Column 3x150 SGX C18 7 µm No. 118 Vosmanska M., Linhart I. Determination of Cotinine by HPLC in Human Urine on CGC Column 3x150 SGX C18

5 µm No. 119 Tesarova E., Sinibaldi M. Separation on Dansyl Derivatives of Amino Acids after Hydrolysis of Dentin from


38

Antiant Teeth on CGC Column 3x150 SGX RPS 5 µm No. 120 Tesarova E., Sinibaldi M. Determination of D/L Ratio of Selected Dansyl Amino Acids from Dentin as a Tool of

Dating of Archeological Sites on CGC Column 3x150 SGX RPS 5 µm No. 121 Vodickova I., Chundela Z. Detection of Mesocarb in Urine on CGC Column 3x150 SGX C18 7 µm No. 122 Vodickova I., Slechtova R., Rehak V. Screening of Diuretics in Human Urine on CGC Column 3x150 SGX C18

7 µm No. 123 Vosmanska M., Linhart I. HPLC Determination of Urinary Carboxylic Acids after Derivatization by 2-

Nitrophenylhydrazine on CGC Column 3x150 SGX C18 5 µm No. 124 Dolezalova M. Determination of Cortisol in Serum on CGC Column 3x150 SGX C18 7 µm No. 125 Drdak M., Simko P. Separation of Elderberry Pigments on CGC Column 3x150 SGX C18 7 µm No. 126 Drdak M., Simko P. Separation of Red Beet Pigments on CGC Column 3x150 SGX C18 7 µm No. 127 Horak T. Determination of Sodium Glutamate in Foodstuffs on CGC Column 3x150 SGX C18 7 µm with

Precolumn CGC 3x30 SGX C18 7 µm No. 128 Ondra P., Husek A. Determination of Neutral Colchicinoid Alkaloids in Plant Extracts on Steel Column 4x250

Tessek Separon SGX C18 7 µm No. 129 Ondra P., Husek A. Determination of Basic Colchicinoid Alkaloids in Plant Extracts on Steel Column 4x250

Tessek Separon SGX C18 10 µm No. 130 Cejpek K., Hajslova J., Kratochvilova B., Merhaut J. Isolation of PAH´s from Fats, Oils and other Lipidic

Extracts on Steel Column 8x500 BIO BEADS S-X3 No. 131 Kroupa F. Determination of Carbofurane in Soil and HopLeaveson CGC Column 3x150 SGX C18 7 µm with

Precolumn 3x30 SGX C18 7 µm No. 132 Jirsa M. Determination of Cyclosporin A in whole Blood on Steel Column 4x250 SGX C18 5 µm No. 133 Rittich B., Spanova A. High Performance Size Exclusion Chromatography Purification of Plasmide DNA Probe

on Steel Column 8x250 HEMA BIO 10 µm No. 134 Svoboda L. Determination of Ammoniacal Nitrogen in Fertilizers CGC Column 3x150 HEMA BIO 1000 SBL 10

µm To obtain a copy of any of these application notes, send an e-mail to [email protected], indicating the number.

Classification Of Sorbents, Powders, and Solid Particles

39

Tessek Ltd. offers classification of HPLC sorbents, solid particles, and powders. Our pilot plant laboratory is equipped with a sophisticated system of ALPINE and ALGAIER classification apparatus operated by skilled. experienced staff. This enables us to classify not only sorbents for HPLC and GC, but also many other powdered materials with particle sizes from 3 - 250 µm. We offer great accuracy, narrow particle size distribution curve, and good yields at reasonable prices. Ranges of typical classification carried out for HPLC sorbents:

Nominal Particle Size (µm) Range (µm) 3 2-4 4 3-5 5 4-6 7 5-9 10 8-12 12 10-14 15 12-18 20 15-27 30 22-38 40 30-50 60 40-80

100 60-140 150 110-190 200 160-240 250 200-300

The particle diameters are evaluated by means of microscopy in a dry state. When desired, materials with larger particles can be disintegrated before classification.

Socochim SA

Ch. du Trabandan 28

1006 Lausanne

Tel +41 (0)21 721 04 50

Fax +41 (0)21 721 04 51

[email protected]

www.socochim.ch

Documents

Mise en page 1 (Page 1) - SocochimHEMA 40, 10 µm 101100005 HEMA 100, 10 µm 101120005 HEMA-S 300, 10 µm 101141005 HEMA-S 1000, 10 µm 101161005 Column STEEL 8x250 mm Cat. Number