vdspher - vds optilab

HPLC . UHPLC . LC-MS . HILIC

HIGH PERFORMANCE SEPARATING PHASES

most accurate analytical results

maximum lifetime

quickest analysis

Improved pressure stability guaranteed

VDSpher

2

…consists of the following strong members: VDSpher, VDSpher PUR, U-VDSpher PUR as well as OptiAqua and OptiBio.

We would like to specify the three VDSpher phases. The VDSpher HPLC phase family was developed for all users who are placing emphasis on straightforward and fast HPLC analysis. The VDSpher HPLC phase family is suitable for both development and quality control in pharmaceutical and food industry. Further applications are in: biochemical research, clinical chemistry and drug monitoring. In short, everyone involved in HPLC analysis will be able to use these phases

High-End End-Capping

excellent batch-to-batch reproducibility

large selection of RP phases

large selection of particle sizes (1.8 μm to 30 μm)

very high degree of purity

Separates what needs to be separated.. .

The VDSpher HPLC separating phases family…

60 Å 60 - 45.5 = 14.5 Å

75 Å 75 - 45.5 = 29.5 Å

100 Å 100 - 45.5 = 54.5 Å

120 Å 120 - 45.5 = 74.5 Å

150 Å 150 - 45.5 = 104.5 Å

200 Å 200 - 45.5 = 154.5 Å

300 Å 300 - 45.5 = 254.5 Å

VDSpher – the material!

3

as an excellent problem solving tool. A wide choice of phase modifications is available to both the analyst and the preparative user.

For analytical HPLC best suitable phases have pore sizes of 100 to 130 Å for analytes with small and medium molecular weights, respectively. VDS optilab decided in favour of 100 Å as standard. For large molecules VDS optilab offers phases from 150 Å to 1000 Å. A 75 Å separating phase is available for preparative applications to the production scale. The small pore size of 75 Å phase recommends a modification with C8 chains (see chart on page 2 and following chart).

VDSpher

Modified Silica

Diagram shows ideal elongated C18 chains at

lipophile eluents (100% organic solvents) and

folded/cumulative, yet also drop shaped C18 chains

at lipophobe eluents (100% aqueous eluent).

It is shown that available pore size of e.g. 60 Å basic

silica is significantly reduced after chemical

modification. In theory, corresponding chart

(on page 2) shows still available pore sizes in

lipophobe condition.

Ads.

Lay

er T

hicknes

s (Å

)

Carbon number

16

14

12

10

8

6

4

2

050 10 2015

VDSpher 75 C18-E is available on request. 75 Å separating phases combine a large surface with a resulting high separating capacity and excellent selectivity.

MeCN

MeOH

THF

4

VDSpher and VDSpher PUR/U-VDSpher PURare based on two differently spherical silica gelsas shown by the data in the following chart:

Narrow particle sizes and pore size distribution guarantee sharp peaks and

create high plate numbers.

VDSpher VDSpher PUR U-VDSpher PUR

Pore size 75 100 150 200 300 1000 100 150 100

Surface m²/g 500 300 175 130 90 30 300 175 300

Pore volume ml/g 0.8 0.8 0.8 0.8 0.8 0.8 0.8 0.8 0.8

Si concentration % 99.95 99.97 99.97 99.97 99.97 99.97 99.995 99.995 99.995

pH value 6.5 6.5 6.5 6.5 6.5 6.5 5.5 5.5 5.5

H2O concentration % <2 <2 <2 <2 <2 <2 <2 <2 <2

Ore grade ppm <125 <100 <100 <100 <100 <100 <60 <60 <60

Density g/ml 0.45 0.45 0.45 0.45 0.45 0.42 0.45 0.45 0.45

Charts 1

Physico-chemical data of the VDSpher separating phases

Sharp peaks, high plate numbers and good select iv ity

Product benefits

High plate numbers and good selectivity are pre-conditions for premium separating performances of VDSpher HPLC columns.

5

In an exemplary way, separating performances of the

VPSpher HPLC columns are reflected with separation

of fat-soluble vitamins... ... and aromatic hydrocarbons

inj

20 1 3 4 5 6

Column 250 x 4.6 mmVDSpher 100 SIL – 5 μm

Conditions

Mobile Phase: n-HexaneFlow rate: 2 ml/min Temperature: ambientDetection: UV 254 nmInjection: 2.0 μl

Sample: 1. Naphthalene 2. Diphenyl 3. Anthracene 4. Pyrene 5. Fluranthene

(min)

1

2

3

45

inj

20 1 3 4 5 6

Column 250 x 4.6 mmVDSpher 100 SIL – 5 μm

Conditions

Mobile Phase: n-Hexane/ Dioxane/Ethanol (97.5/2.0/0.5)Flow rate: 1.5 ml/min Temperature: ambientDetection: UV 295 nmInjection: 12 μl

Sample : 1. -Tocopherol 2. -Tocopherol 3. -Tocopherol 4. -Tocopherol

(min)

1

2

3

4

100

75

50

25

0

Pore diameter (nm)

101 100

75 Å

100 Å

300 Å

Particle Size Distribution

Application Aromatic Hydrocarbons

Pore Size Distribution

Application Vitamin E

Cumulative (%) Cumulative (%)

Particle Size (μm)

1009080706050403020100

5 10 15 20 25 30

2 4 6 8 10 15 20 25 30

1 2 3 1 5 μm

2 10 μm

3 15 μm

VDSpher

6

In addition to the standard silica VDSpher SIL, ultrapure VDSpher PUR SIL and U-VDSpher PUR SIL materials are available. Both silica gels are characterised by a high

Silica phases are the basis of reversed phases (RP), normal phases (NP), ion exchange phases (IAP) and HILIC phases.

VDSpher SIL separating phases

Pore Size Particle Size Surface Area USP

75 Å 5/10/15 300 m2/g L3

100 Å 5/10/15 300 m2/g L3

150 Å 5 175 m2/g L3

200 Å 5 130 m2/g L3

300 Å 5/10/15/30 90 m2/g L3

1000 Å 5 30 m2/g L3

VDSpher PUR SIL separating phases

Pore Size Particle Size Surface Area USP

100 Å 2.2/3/3.5/4/5/10 300 m2/g L3

150 Å 5 175 m2/g L3

U-VDSpher PUR SIL separating phases

Pore Size Particle Size Surface Area USP max. Pressure

100 Å 1.8 300 m2/g L3 800 bar

Al l phases with high degree of purity

degree of purity (see chart 1, page 4). List of available phases as shown in chart 2 show corresponding particle sizes.

Charts 2

Silica phases

7

Entire RP program contains VDSpher, ultrapure VDSpher PUR as well as U-VDSpher PUR for UHPLC applications. As shown by the phase tree, short-chained to long-chained modifications appear in the reversed phase

Reversed phases (RP phases)

VDSpher

area. In addition to the standard C18-E phase, another five different C18 modifications are available, from hydrophilic (C18-H) to hydrophobic (C18-M-SE), with pore sizes from 75 Å to 300 Å.

normal phase

CN HILIC

PEI

aqueous aqueous/organic organic

Diol HILIC-OH OptiAqua H E

NH2

HILIC-AMM

not end-capped

OptiBio

SE

M – E

M – SE

HILIC phase C18

SB

E

E

E SE

SE

H

OptiBio

OptiBio

C8 Phenyl C4

VDSpher silica

Ion exchange

phase

8

Different separating propert ies under the same separating condit ions

Different available modifications are shown with C18 phases in an exemplary manner:

A 40% surface coverage is achieved with liquid phase endcapping. Gas phase endcapping gains up to 99% surface coverage.

Following chromatograms (on page 9) show different separating properties of C18-E (hydrophobic surface) and C18-H (hydrophilic surfaces) under same separating conditions.

C18-E Monomeric bonding Liquid phase endcapping

C18-SE Monomeric bonding Gas phase endcapping

C18-M Polymeric bonding no endcapping

C18-M-E Polymeric bonding Liquid phase endcapping

C18-M-SE Polymeric bonding Gas phase endcapping

C18-H Monomeric bonding Polar spacer

9

VDSpher

0 5 10 15 20

Column 150 x 4.6 mm

VDSpher 100 C18-H – 5 μm

Column 150 x 4.6 mm

VDSpher 100 C18-E – 5 μm

Conditions

Mobile Phase: Methanol/Water

(80/20)

Flow rate: 1.0 ml/min

Temperature: 36 °C

Detection: UV 254 nm

Injection: 10 μl

Sample: 1. Uracil

2. Ethylbenzene

3. n-Propylbenzene

4. n-Butylbenzene

5. n-Amylbenzene

Times (min)

1

2

3 45

VDSpher 100 C18-H versus VDSpher 100 C18-E

C18-E

0 5 10 15 20

Times (min)

1

2

3

45

C18-H

10

Benefit ing from different surface chemistry

At approximately same C concentrations, C18-H phase shows different separating properties than a C8-E phase. Different surface chemistry of phases is responsible for this. Bonded polar component of C18-H has a comparable length as the RP chain. Adjustment of retention times can be observed with preparative clean up of insulin.

Modification C18-H is a very hydrophilic phase that can be used with both 100% water and buffer agent.

Standard phase C18-E is the first option for many separating problems. With this phase, e.g. determination of caffeine in different beverages is possible (e.g. coffee, tea, cola or energy drinks).

0 05 510 1015 1520 20 25

Times (min) Times (min)

1

1

2

2

3

3

Two different RP-Surface Chemistries by same C value

VDSpher 100 C8-E – 10 μm VDSpher 100 C18-H – 10 μm

Sample: Purifikation of Insulin with preparative HPLC (production scale)

10.2 % C 10.0 % C

11

VDSpher

Phase C18-SE is also perfectly suitable for a wide range of separating problems. Phase C18-SE is characterised by a special double end-capping and is of medium hydrophobicity, however slightly more hydrophobic than C18-E. Especially with hydrophobic substances, a larger retention can be observed than with C18-E. Based on the density (up to 99%), phase C18-SE shows improved stability with acids and bases.

Highly hydrophobic phase C18-M-SE is also offered, probably one of the most hydrophobic phases on the market. With this phase, e.g. additives of soft drinks can be analysed.The C18-M phase is water-compatible. Excellent shielding of the silica surface makes this possible – in spite of stronger hydrophobicity. C18-M is also a great option for LC/MS analytics as there is no „carry-over effect“.

Furthermore, phases program includes short-chained RP phases with C8 and C4 modifications. With these phases, lower hydrophobicity in

connection with residual silanol activity is jointly responsible for the separating mechanism. A special feature is the phenyl phase with - interaction with aromatic eluents. This phase is also endcapping.

Here, by way of example, test chromatogram of a VDSpher C18-SE column. Test certificates are documenting quality of each produced column:

0 1 2 3 4 5 6 7

Times (min)

1

2

3

Column 150 x 4.0 mmVDSpher 100 C18-SE – 5 μm

Conditions

Mobile Phase: CH3CN/H

2O (75:25)

Flow rate: 1 ml/min Temperature: ambientDetection: UV 254 nmInjection: 5.0 μl test mixture RP

Sample: 1. Benzene 2. Naphthalene 3. Anthracene

C18-SE test chromatogram

N = 120.226/m

12

VDSpher RP separating phases

Phase Pore Particle C % Endcapping PH Surface USP Size Size Range Area

C18-E 75 Å 5 20.8 yes 2 - 7.5 500 m2/g L1

C18-E 100 Å 5/10/15/30 16.2 yes 2 - 7.5 300 m2/g L1

C18-E 150 Å 5 10.5 yes 2 - 7.5 175 m2/g L1

C18-E 200 Å 5/10 8.5 yes 2 - 7.5 130 m2/g L1

C18-E 300 Å 5/10/15 6.9 yes 2 - 7.5 90 m2/g L1

C18-E 300 Å 30 5.8 yes 2 - 7.5 90 m2/g L1

C18-H 100 Å 5/10 10.0 polarer Spacer 3 - 7.5 300 m2/g L1

C18-SE 100 Å 5/10 16.5 yes (special) 2 - 9 300 m2/g L1

C18-M 100 Å 5/10 17.5 no 2 - 7.5 300 m2/g L1

C18-M-E 100 Å 5/10 19.5 yes 2 - 7.5 300 m2/g L1

C18-M-SE 100 Å 5/10 20.0 yes (special) 2 - 9 300 m2/g L1

C8-E 75 Å 5 13.8 yes 2 - 7.5 500 m2/g L7

C8-E 100 Å 5/10 10.2 yes 2 - 7.5 300 m2/g L7

C8-H 100 Å 5/10 8.0 polarer Spacer 3 - 7.5 300 m2/g L7



C8-SB 75 Å 5/10 13.6 yes (special) 2 - 8.5 500 m2/g L7

C8-SB 100 Å 5/10 10.5 yes (special) 2 - 8.5 300 m2/g L7

C4-E 100 Å 5/10 7.0 yes 2 - 7.5 300 m2/g L26

C4-E 300 Å 5/10 2.6 yes 2 - 7.5 90 m2/g L26

C4-E 300 Å 15 2.2 yes 2 - 7.5 90 m2/g L26

C4-SE 100 Å 5 7.1 yes (special) 2 - 9 300 m2/g L26

Phenyl-E 100 Å 5/10 12.2 yes 2 - 7.5 300 m2/g L11

Charts 3

RP phases

RP separating phases of the VDSpher product ser ies at a glance

13

VDSpher

VDSpher PUR RP separating phases


C18-E 100 Å 2.2/3/3.5 16.2 yes 2 - 7.5 300 m2/g L1 4/5/10

C18-H 100 Å 2.2/3/5/10 10.0 polarer 3 - 7.5 300 m2/g L1 Spacer

C18-SE 100 Å 3.5/4/5/10 16.5 yes (special) 2 - 9 300 m2/g L1

C18-M 100 Å 2.2/3/3.5/ 17.5 no 2 - 7.5 300 m2/g L1 4/5/10

C18-M-E 100 Å 3.5/4/5/10 19.5 yes 2 - 7.5 300 m2/g L1

C18-M-SE 100 Å 3.5/4/5 20.0 yes (special) 2 - 9 300 m2/g L1

C8-E 100 Å 2.2/3/3.5 10.2 yes 2 - 7.5 300 m2/g L7 4/5/10

C4-E 100 Å 2.2/3/3.5/4/5 7.0 yes 2 - 7.5 300 m2/g L26

C4-SE 100 Å 5 7.1 yes (special) 2 - 9 300 m2/g L26

Phenyl-E 100 Å 3.5/4/5 12.2 yes 2 -7.5 300 m2/g L11

U-VDSpher PUR RP separating phases

Phase Pore Particle C % Endcapping PH Surface USP max. Size Size Range Area Pressure

C18-E 100 Å 1.8 16.2 yes 2 - 7.5 300 m2/g L1 800 bar

C18-H 100 Å 1.8 10.0 polarer 3 - 7.5 300 m2/g L1 800 bar Spacer

C8-E 100 Å 1.8 10.2 yes 2 - 7.5 300 m2/g L7 800 bar

Phenyl-E 100 Å 1.8 12.2 yes 2 -7.5 300 m2/g L11 800 bar

14

Reproducible analyt ics by means of fast adjustment of the phase balance

Besides silica phases (SIL) the following modifications are available:

CN, Diol and NH

2

Normalphasen (NP phases)

Advantages of chemically-bonded NP phases are:

fast adjustment of the phase balancereproducible analytics despite slightly varying

water contents (in the ppm area). Reproducibility can be additionally improved

by adding a more polar modifier (e.g. dioxane)

0 0.50 1.00 1.50 2.00 2.50 3.00 3.50

Times (min)

1

2

Column 150 x 4.6 mmVDSpher 100 CN – 5 μm

Conditions

Mobile Phase: n-Heptane/Dioxane (9:1)Flow rate: 1 ml/min Temperature: ambientDetection: UV 254 nmInjection: 5.0 μl test mix NP

Sample: 1. Benzene 2. Nitrobenzene

Test chromatogram VDSpher CN

N = 119.006/m

Product benefits

15

VDSpher

VDSpher normal phases


CN 100 Å 5/10 6.5 no 2 - 8 300 m2/g L10

Diol 100 Å 5 4.1 no 2 - 8 300 m2/g L20

NH2 100 Å 5/10 3.8 no 2 - 7.5 300 m2/g L8

VDSpher PUR normal phases


CN 100 Å 3.5/4/5 6.5 no 2 - 8 300 m2/g L10

Diol 100 Å 5 4.1 no 2 - 8 300 m2/g L20

NH2 100 Å 2.2/3/3.5/4/5 3.8 no 2 - 7.5 300 m2/g L8

U-VDSpher PUR normal phases

Phase Pore Particle C % Endcapping PH Surface USP max. Size Size Range Area Pressure

CN 100 Å 1.8 6.5 no 2 - 8 300 m2/g L10 800 bar

Charts 4

RP phases

16

New: optimum exchanger capacity in the pH range from 4 to 7

Besides the standard NH2 phase as weak

anion exchanger the new PEI phase (PolyEtylenImin) is also available as weak basic anion exchanger.

Ion exchange phase

The phase reaches its optimum exchange capacity in the pH range from 4 to 7. Ionic strength and pH changes of the mobile phase are responsible for the separation, i.e. reversible adsorption of negatively charged analytes at positively charged exchange groups (PEI) is disbanded.

0 0.50 1.00 1.50 2.00 2.50 3.00

Times (min)

1

2

Column 125 x 4.0 mmVDSpher PUR 100 PEI – 5 μm

Conditions

Mobile Phase: n-Heptane/Dioxane (9:1)Flow rate: 1 ml/min Temperature: ambientDetection: UV 254 nmInjection: 5.0 μl test mix NP

Sample: 1. Benzene 2. Nitrobenzene

Test chromatogram PEI

17

VDSpher

VDSpher PUR PEI

Phase Pore Particle C % Endcapping PH Surface Size Size Range Area

PEI 100 Å 5 7.5 no 4 - 7 300 m2/g

Charts 5

Ion exchange phase

18

Part it ion chromato-graphy between polar stat ionary and polar mobi le phases

Many phases can be used in HILIC mode as well. Besides the adequate silica phase, VDS optilab GmbH also offers specifically selected phases

HILIC-AM and HILIC-OH.

In this context, amino phase can be regarded as classic. Sugar separations by means of this phase have been known for a long time. Separation in HILIC mode is carried out as separation between the polar stationary phase and the also polar mobile phase. Particularly strong polar analytes that are very strongly retarded on RP phases can be perfectly analysed.

HILIC phases

VDSpher PUR HILIC

Phase Pore Particle C % Endcapping PH Surface Size Size Range Area

HILIC 100 Å 5 no 2 - 8 300 m2/g

HILIC-AM 100 Å 5 3.8 no 2 - 8 300 m2/g

HILIC-OH 100 Å 5 4.0 no 2 - 8 300 m2/g

Charts 6

HILIC phases

19

Asia Pacific

IndiaINKARP Instruments Private Ltd.1-2-45/1 Street No. 2Kakatiya Nagar Colony, HabsigudaHyderabad - 500 007Phone: +91 40 27172293 + 27175088Fax: +91 40 [email protected]

Middle East and Africa

EgyptGroup Engineering & Scientific SystemsHouse 18, Str 219, Al MaadiPOB 1024 Al Maadi CairoPhone: +202 25197043Fax: +202 [email protected]

Europe

GermanyVDS optilabChromatographietechnik GmbHWiesenweg 11aD 10365 BerlinPhone: ++49 (0)30 5515 3901Fax: ++49 (0)30 5515 [email protected]

FinlandKrotek OyRantatie 1133250 TamperePhone: +358 3 2535218Fax: +358 3 [email protected]

VDSpher

KoreaWoong Ki Science Co., Ltd277-38 Sungsu 2gaSungdong-GuSeoul, KoreaPhone: +82-2-461-3811Fax: [email protected]

IsraelM. Snir Technologic Services Ltd.P.O. Box 10914M. Haifa 26119Phone: +972 4 8438991/2Fax: +972 4 [email protected]

ItalyItalscientifica S.r.l.Via Assarotti 5/616122 GenovaPhone: +39 10 8424261Fax: +39 10 [email protected]

AustriaHPLC ServiceDr. Manfred Wagner-Löffler Franziska Lechnergasse 24 2384 BreitenfurtPhone: +43 2239 4271Fax: +43 2239 [email protected]

America

MexicoLEACSA, S.A de C.V. Alfonso Esparza Oteo #63, Col. Guadalupe InnC.P. 01020 México, D.F.Phone: +55 56620010Fax: +55 [email protected]

TaiwanScientific Hightek Co., Ltd10F-5, No. 21, Sec. 6ZhongXiao E. Road11575 Taipei, TaiwanPhone: +886 2 26522200Fax: +886 2 [email protected]

MoroccoScomedica22 Rue Mausolée, Résidence AbirQuartier des Hôpitaux20100 CasablancaPhone: +212 22 864578Fax: +212 22 [email protected]

PortugalVia Athena Gestáo de Laboratórios, Lda.Rua Fábrica da Loiça de Sacavém, nº3-B2685-135 SacavémPhone: +351 219 409 940Fax: +351 219 409 [email protected]

HungaryKvalitex Kft.Pannónia U. 5.1136 BudapestPhone: +36 1 3404700Fax: +36 1 [email protected]

Autorized distribution partners of VDS optilab

© VDS optilab, Chromatographietechnik GmbH

VDS optilabChromatographietechnik GmbH

Wiesenweg 11aD 10365 Berlin

Phone: ++49 (0) 30 55 15 39 01Fax: ++49 (0) 30 55 15 39 03

e-mail: [email protected]: www.vdsoptilab.de

Made in Germany

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