Columns for HPLC - Western Analytical

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Fast LC of anilinesColumn: 30 x 4 mm NUCLEOSIL ® 100-5 PROTECT IEluent: methanol – water(45:55, v/v)2Flow rate: 2 ml/minDetection: UV, 254 nmPeaks:11. Aniline2. N,N-Dimethylaniline3. N,N-Diethylaniline3RP method development kitsFor selection of the RP phase best suited for your separationwe offer two RP method development kits with 30 mmChromCart ® columns:each kit consists of a CC column holder 30 mm and one columneach 30 x 2 or 30 x 4 mm, respectively, packed with:NUCLEOSIL ® PROTECT I, NUCLEOSIL ® 100-5 C 18 HD,NUCLEOSIL ® 100-5 C 8 HD, NUCLEOSIL ® 100-5 C 18 ABand NUCLEOSIL ® 100-5 C 18Ordering informationDesignationCat. No.Method development kit ChromCart ® 2 mm 721125.20Method development kit ChromCart ® 4 mm 721125.40Method development kit ChromCart ® 4.6 mm 721125.461149800 1 2 minwww.mn-net.comComparison of selectivities for base deactivated RP phasesSeparation of a test mixture on four base deactivated NUCLEOSIL ® phasesThese chromatograms show the differences in retention times and selectivities of the four phases.Columns: A) 250 x 4 mm NUCLEOSIL ® 100-5 C 18 HD; B) 250 x 4 mm NUCLEOSIL ® 100-5 C 18 AB;C) 250 x 4 mm NUCLEOSIL ® 100-5 C 8 HD; D) 250 x 4 mm NUCLEOSIL ® 100-5 PROTECT IEluent: methanol – water (60:40); flow rate 0.8 ml/min; temperature: 35 °CDetection: UV, 230 nmPeaks:1. p-Ethylaniline2. Diethyl phthalate3. N,N-Dimethylaniline4. Ethyl benzoate5. Toluene6. Naphthalene7. EthylbenzeneC 18 HD C 18 AB C 8 HD PROTECT2,32,3133 444246121515755766766.7.050 10 20 min0 10 20 30min0 10 20 min 0 10 min24www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Base deactivated phases for RP chromatography: NUCLEOSIL ® HDNUCLEOSIL ® HD is a family of RP phases with optimisedbase deactivation, which is available with octyl or octadecylmodification and 3 or 5 µm particles.NUCLEOSIL ® 100-5 C 8 HD is an octyl modified RP phasewith 5 µm particles. As can be seen in the figure below, thebasic compound lidocaine is eluted without tailing. Comparedto NUCLEOSIL ® 100-5 C 18 HD introduction of theshorter carbon chain results in a reduction of hydrophobic interactions.The chromatograms show the separation of compounds withdifferent functional groups on both phases. The less polar aromaticstoluene and naphthalene are eluted earlier from theC 8 HD phase resulting in a reversed elution sequence of tolueneand lidocaine.www.mn-net.comComparison of selectivities between NUCLEOSIL ® HD octadecyl and octyl phasesSeparation of a test mixture on NUCLEOSIL ® C 8 HDColumn: 250 x 4 mm NUCLEOSIL ® 100-5 C 8 HDEluent: MeOH – 25 mM NaH 2 PO 4 pH 7.0 (65:35)Flow rate: 0.8 ml/minTemperature: 25 °CDetection: UV, 254 nm13Peaks:1. Phenol242. 5’-(p-Methylphenyl)-5-phenylhydantoin3. Diethyl phthalate4. Lidocaine55. Toluene6. Naphthalene6110000Separation of a test mixture on NUCLEOSIL ® C 18 HDColumn: 250 x 4 mm NUCLEOSIL ® 100-5 C 18 HDEluent: MeOH – 25 mM NaH 2 PO 4 pH 7.0 (65:35)Flow rate: 0.8 ml/minTemperature: 25 °CDetection: UV, 254 nmPeaks:1. Phenol1 32. 5’-(p-Methylphenyl)-425-phenylhydantoin3. Diethyl phthalate4. Lidocaine55. Toluene6. Naphthalene61106500 5 10 15 min0 10 20 minHighest quality! · Batch-to-batch reproducibility of NUCLEOSIL ® HDSeparation of a test mixture on three different batchesof NUCLEOSIL ® 100-5 C 8 HDColumn 250 x 4 mm NUCLEOSIL ® 100-5 C 8 HDEluent methanol – water (55:45, v/v); flow rate 1 ml/mintemperature 40 °C; detection: UV, 254 nm2Peaks:1. Thiourea42. Aniline13. Phenol54. o,m,p-Toluidine36 75. N,N-Dimethylaniline6. Methyl benzoate7. Toluene8 8. Ethylbenzene0 10 20 min11089086420LotACapacity factork’ (toluene)k’ (ethyl benzoate)k’ (MPPH)BCα (MPPH/toluene)α (ethyl benzoate/toluene)DEF7654321SelectivityNUCLEOSIL ® 100-5 C 18 HD, acetonitrile – water 50:50(v/v), 1 ml/min, 25 °C, UV 254 nm6.7.05MNwww.mn-net.com27

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyNUCLEOSIL ® 100-3 C 8 HDOctyl 3 µmspherical 3 µm silica particles with monomeric octylmodificationNUCLEOSIL ® 100-5 C 8 HDOctyl 5 µmspherical 5 µm silica particles with monomeric octylmodificationNUCLEOSIL ® 100-3 C 18 HDOctadecyl 3 µmspherical 3 µm silica particles with monomeric octadecylcoatingNUCLEOSIL ® 100-5 C 18 HDOctadecyl 5 µmspherical 5 µm silica particles with monomeric octadecylcoatingThese packings are manufactured from NUCLEOSIL ® 100with 3 or 5 µm particles, respectively. Thus, surface shielding,capacity factors and selectivities are the same for both materials.However, the number of theoretical plates is higher whenusing the 3 µm starting silica. The 3 µm particle allows applicationof shorter columns (= reduction of time required for theanalysis) and improves the detection sensitivity due to smallerpeak widths.The following chromatogram exhibits the higher efficiency ofthe 3 µm NUCLEOSIL ® C 8 HD column compared to the 5µmequivalent for identical column lengths (curves a and b). Theimproved sensitivity of the 3 µm packing becomes evident bythe increased peak heights in the chromatogram. However, ifthe column length is reduced from 250 mm to 150 mm, baselineseparation, under the given chromatographic conditions,cannot be achieved (c). In this case the use of the 250 mmNUCLEOSIL ® 100-3 C 8 HD column is strongly recommendedto obtain optimal results.Separation of paracetamol, caffeine and acetylsalicylic acidSample preparation:1dissolve 1 tablet in 20 ml ACN/water (50:50), dilute 1:10.Eluent: ACN – water – H 3 PO 4 (20:80:0.1, v/v/v)Columns: a) 250 x 4 mm NUCLEOSIL ® 100-5 C 8 HDTemperature: 30 °Cb) 250 x 4 mm NUCLEOSIL ® 2100-3 C 8 HDFlow rate: 1 ml/minc) 150 x 4 mm NUCLEOSIL ® 100-3 C 8 HDDetection: UV, 254 nmPeaks:1. Paracetamol2. Caffeine3. Acetylsalicylicacid31169606.7.052 4 6 8 10 12 2 4 6 8 10 12 2 4 6 8 minOrdering information · Analytical columns with NUCLEOSIL ® HDLength → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-3 C 8 HDParticle size 3 µm, pore size 100 Å; octyl phase, endcapped, monomer coating, 13% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721656.20 721657.20 721658.20 721669.20 721612.303 mm ID 721656.30 721657.30 721658.30 721669.30 721612.304 mm ID 721656.40 721657.40 721658.40 721669.40 721612.404.6 mm ID 721656.46 721657.46 721658.46 721704.46 721669.46 721612.40Microbore columns 2)1 mm ID 717115.10 717116.10 717117.10 717118.10EC columns 3)2 mm ID 720526.20 720528.20 721612.303 mm ID 720526.30 720528.30 721612.304 mm ID 720526.40 720528.40 721612.404.6 mm ID 720526.46 720527.46 720528.46 721612.40Each column is individually tested and supplied with test chromatogram and test conditions28www.mn-net.comMN

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyOrdering information · Analytical columns with NUCLEOSIL ® HDLength → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 C 8 HDParticle size 5 µm, pore size 100 Å; octyl phase, endcapped, monomeric coating, 13% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721071.20 721499.20 721497.20 721498.20 721500.303 mm ID 721071.30 721499.30 721497.30 721498.30 721500.304 mm ID 721071.40 721499.40 721497.40 721498.40 721500.404.6 mm ID 721071.46 721499.46 721497.46 721501.46 721498.46 721500.40Microbore columns 2)1 mm ID 717043.10 717046.10 717049.10 717057.10EC columns 3)2 mm ID 720195.20 720196.20 721500.303 mm ID 720195.30 720196.30 721500.304 mm ID 720195.40 720196.40 721500.404.6 mm ID 720195.46 720194.46 720196.46 721500.40NUCLEOSIL ® 100-3 C 18 HDParticle size 3 µm, pore size 100 Å; octadecyl phase, endcapped, monomeric coating, 20% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721196.20 721493.20 721491.20 721492.20 721494.303 mm ID 721196.30 721493.30 721491.30 721492.30 721494.304 mm ID 721196.40 721493.40 721491.40 721492.40 721494.404.6 mm ID 721196.46 721493.46 721491.46 721495.46 721492.46 721494.40Microbore columns 2)1 mm ID 717037.10 717038.10 717039.10 717040.10EC columns 3)2 mm ID 720191.20 720192.20 721494.303 mm ID 720191.30 720192.30 721494.304 mm ID 720191.40 720192.40 721494.404.6 mm ID 720191.46 720193.46 720192.46 721494.40NUCLEOSIL ® 100-5 C 18 HDParticle size 5 µm, pore size 100 Å; octadecyl phase, endcapped, monomeric coating, 20% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721072.20 721851.20 721852.20 721850.20 721853.303 mm ID 721072.30 721851.30 721852.30 721850.30 721853.304 mm ID 721072.40 721851.40 721852.40 721850.40 721853.404.6 mm ID 721072.46 721851.46 721852.46 721854.46 721850.46 721853.40Microbore columns 2)1 mm ID 717033.10 717024.10 717015.10 717001.10EC columns 3)2 mm ID 720296.20 720280.20 721853.303 mm ID 720296.30 720280.30 721853.304 mm ID 720296.40 720280.40 721853.404.6 mm ID 720296.46 720294.46 720280.46 721853.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) 30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other lengths are available on requestMNwww.mn-net.com29

OHH OHHOO HHColumns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyNUCLEOSIL ® 100 C 18 NautilusPolSiSithe formula for RP chromatography in aqueous mediaWith highly aqueous mobile phases (> 95 %) conventionalreversed phase columns very often exhibit a plainly visibledeterioration of column performance after a certain period oftime. The non-polar alkyl chains lose their "brush-type structure"for maintaining the hydrophobic interactions betweenstationary phase and analyte. The result of this phase collapseis a drastic decrease of retention time and poor resolution.This phenomenon may occur very fast or after a while.H 2 OH 2 OAAH 2 OH 2 OAH 2 OH 2 OH 2 OAH 2 OH 2 OH 2 OAH 2 OH 2 OAH 2 OH 2 OA = analyteAH 2 OAH 2 OAEven after days or weeks of operation in purely aqueous eluentsthe C 18 chains of NUCLEOSIL ® 100-5 C 18 Nautilus neitherare folded nor show any collapsing. A significant reductionof retention time cannot be observed as is shown in thefollowing diagrams.% of initial retention time10080604020Phase collapse in aqueous mediachange of retention time with 100% aqueous eluentsNUCLEOSIL ® 100-5 C 18 Nautilusconventional RP phaseday00 2 4 6 86.7.05Start2468Phase collapse and decrease in retention times for a conventionalRP phase with 100 % aqueous eluentsNUCLEOSIL ® 100-5 C 18 Nautilus is a reversed phase C 18column based on highly pure silica with thorough endcapping,which is totally stable if the mobile phase is 100%aqueous without any organic modifiers. This property isachieved by a covalently bonded C 18 silane (synthesised inMACHEREY-NAGEL’s R&D laboratories) with an embeddedfunctional group, which is the driving force for the excellentwater stability of the RP surface. Compared to standard C 18phases where hydrophobic interactions between analyte andstationary phase predominate, NUCLEOSIL ® 100-5 C 18Nautilus also shows polar interaction via hydrogen bondingactivities and dipole-dipole forces. This can influence retentiontime (t R ) and improve selectivity (α) particularly for theseparation of polar compounds.Start2461 23454 8 minSeparation of nucleobases in apurely aqueous eluentColumn: 125 x 4 mm NUCLEOSIL ®100-5 C 18 NautilusEluent: 20 mM ammonium formate,pH 4.1Flow rate: 1 ml/minTemperature: 20 °CDetection: UV, 254 nmPeaks:1. Cytosine2. Uracil3. Hypoxanthine4. Xanthine5. AdenineIn spite of the polar character of the embedded functionalgroup NUCLEOSIL ® C 18 Nautilus exhibits sufficient hydrophobicproperties and is very well suited for analysing basiccompounds just like NUCLEOSIL ® HD or NUCLEOSIL ®Protect I. It may be worthwhile to try this chemistry for difficultseparations.11564030Each column is individually tested and supplied with test chromatogram and test conditionswww.mn-net.comMN

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyIn the selectivity test on the right the strong base p-ethylanilineis eluted as a sharp signal with remarkably good peakshape. Another example for the excellent base deactivationof NUCLEOSIL ® Nautilus is the tailing-free elution of pyridine.Chromatographic conditions:Column 250 x 4 mm NUCLEOSIL ® 100-5 C 18 Nautilus; eluentmethanol – water (left 55:45, right 30:70, v/v); flow rateleft 0.8 ml/min, right 1 ml/min; temperature 40 °C; detectionUV, 254 nmPeaks (left): 1. uracil, 2. aniline, 3. phenol, 4. p-ethylaniline,5. N,N-dimethylaniline, 6. toluene, 7. ethylbenzeneReferencesD. Rieger, H. Riering, Int. Laboratory, Vol. 30, p. 41Selectivity test1 234567NPyridine/phenol4 8 12 16 20 24 28 min 4 8 12 16OHOrdering information · Analytical columns with NUCLEOSIL ® 100-5 C 18 NautilusLength → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-3 C 18 NautilusParticle size 3 µm, pore size 100 Å; octadecyl phase, endcapped, embedded polar group, 16% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721649.20 721650.20 721651.20 721652.20 721611.303 mm ID 721649.30 721650.30 721651.30 721652.30 721611.304 mm ID 721649.40 721650.40 721651.40 721652.40 721611.404.6 mm ID 721649.46 721650.46 721651.46 721703.46 721652.46 721611.40Microbore columns 2)1 mm ID 717110.10 717111.10 717112.10 717113.10EC columns 3)2 mm ID 720472.20 720470.20 721611.303 mm ID 720472.30 720470.30 721611.304 mm ID 720472.40 720470.40 721611.404.6 mm ID 720472.46 720471.46 720470.46 721611.40NUCLEOSIL ® 100-5 C 18 Nautilus as above, however, particle size 5 µmChromCart ® columns2 mm ID 721133.20 721134.20 721131.20 721130.20 721140.303 mm ID 721133.30 721134.30 721131.30 721130.30 721140.304 mm ID 721133.40 721134.40 721131.40 721130.40 721140.404.6 mm ID 721133.46 721134.46 721131.46 721132.46 721130.46 721140.40Microbore columns 2)1 mm ID 717066.10 717065.10 717067.10 717068.10EC columns 3)2 mm ID 720430.20 720431.20 721140.303 mm ID 720430.30 720431.30 721140.304 mm ID 720430.40 720431.40 721140.404.6 mm ID 720430.46 720432.46 720431.46 721140.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) 30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other lengths are available on requestMNwww.mn-net.com31

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyNUCLEOSIL ® 100-3 Protect ISpecial RP phase 3 µmspherical 3 µm silica particles with monomeric specialRP modificationNUCLEOSIL ® 100-5 Protect ISpecial RP phase 5 µmspherical 5 µm silica particles with monomeric specialRP modificationAlthough hydrophobic interactions of NUCLEOSIL ® 100-5Protect I are comparatively low and closer to the standardoctyl phase NUCLEOSIL ® 100-5 C 8 , the special surfacemodification combined with a rigorous endcapping of the unreactedsilanol groups reduces interaction successfully andensures excellent peak shapes with basic compounds.In comparison with the well-established NUCLEOSIL ® 100-5C 18 AB, NUCLEOSIL ® 100-5 C 18 HD and NUCLEOSIL ®100-5 C 8 HD a key characteristic of the NUCLEOSIL ® 100-5protect I is its higher polarity. Nevertheless, critical compoundslike amines show outstanding peak symmetries. Thisstationary phase is well suited for the separation of low molecular-weightcompounds (up to 5000 dalton), e. g. drugs orpesticides.For example, the tricyclic amines protriptyline and amitriptylineshow especially strong interactions with residual silanolgroups on a conventional C 18 column, while ourNUCLEOSIL ® 100-5 Protect I features✔ early elution and✔ excellent peak symmetryfor this critical pair of compounds (see chromatogram).The batch test underlines the outstanding base deactivationand unique selectivity of NUCLEOSIL ® 100-3 Protect I forthe successful separation of critical bases such as amitriptylineand protriptyline, which are eluted with excellent peakshapes as for the aromatic hydrocarbons.Separation of a batch test mixtureColumn: 250 x 4 mm NUCLEOSIL ® 100-3 Protect I,Cat. No. 720542.40Eluent: acetonitrile – 25 mM KH 2 PO 4 , pH 7.5 (50:50)Flow rate: 1 ml/minDetection: UV, 254 nmTemperature: 25 °C2Peaks:31. Uracil2. Protriptyline3. Amitriptyline4. Ethyl benzoate5. Toluene6. Ethylbenzene4116970154 8 12 minwww.mn-net.com66.7.0532Each column is individually tested and supplied with test chromatogram and test conditionswww.mn-net.comMN

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyOrdering information · Analytical columns with NUCLEOSIL ® 100-5 Protect ILength → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-3 Protect IParticle size 3 µm, pore size 100 Å; special RP phase, endcapped, monomer coating, 11% C;eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721670.20 721671.20 721672.20 721673.20 721613.303 mm ID 721670.30 721671.30 721672.30 721673.30 721613.304 mm ID 721670.40 721671.40 721672.40 721673.40 721613.404.6 mm ID 721670.46 721671.46 721672.46 721705.46 721673.46 721613.40Microbore columns 2)1 mm ID 717120.10 717121.10 717122.10 717123.10EC columns 3)2 mm ID 720540.20 720542.20 721613.303 mm ID 720540.30 720542.30 721613.304 mm ID 720540.40 720542.40 721613.404.6 mm ID 720540.46 720541.46 720542.46 721613.40NUCLEOSIL ® 100-5 Protect IParticle size 5 µm, pore size 100 Å; special RP phase, endcapped, monomer coating, 11% C;eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721157.20 721152.20 721151.20 721150.20 721154.303 mm ID 721157.30 721152.30 721151.30 721150.30 721154.304 mm ID 721157.40 721152.40 721151.40 721150.40 721154.404.6 mm ID 721157.46 721152.46 721151.46 721153.46 721150.46 721154.40Microbore columns 2)1 mm ID 717034.10 717025.10 717016.10 717007.10EC columns 3)2 mm ID 720175.20 720170.20 721154.303 mm ID 720175.30 720170.30 721154.304 mm ID 720175.40 720170.40 721154.404.6 mm ID 720175.46 720174.46 720170.46 721154.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05MNwww.mn-net.comCustom-packed columns with other lengths are available on request33

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyApplications4Separation of phenolsColumn: 250 x 4 mm NUCLEOSIL ® 100-5 Protect IEluent: acetonitrile – 25 mM KH 2 PO 4 pH 7 (50:50, v/v)Flow rate: 1 ml/minTemperature: 20 °CDetection: UV, 235 nmPeaks:1. p-Benzoquinone2. Phenol3. o-Nitrophenol4. p-Nitrophenol5. 2,3-Dinitrophenol3 66. m-Nitrophenol910 7. 2,5-Dinitrophenol8. 3,4-Dinitrophenol9. 2,6-Dinitrophenol110. 2,4-Dinitrophenol7 82 511. 2,4,6-Trinitrophenol11111540Separation of sunscreen agentsV. Vanquerp et al.J. Chromatogr. 832 (1999) 273 – 277Column: 250 x 4 mm NUCLEOSIL ®100-5 C 8 HDEluents: A) 1% acetic acidin water,B) methanolGradient: 80 – 100% Bin 10 minFlow rate: 1 ml/minDetection: UV, 300 nmPeaks(Injection volume 10 µl):A) Benzophenone-3AB) Methylbenzylidene camphorC) Octyl methyl PABAD) Butyl methoxydibenzoylmethaneE) PEG-25 PABABCDE1147400 10 20 min0 5 10 15 2011018012Analysis of customary heroin, stretched withparacetamol/caffeineColumn 250 x 2 mm NUCLEOSIL ® 100-5 C 18 AB, withguard column 8 x 3 mm NUCLEOSIL ® 100-5 C 18 ABInjection volume 30 µlEluents: A) 2 mmol NaH 2 PO 4 , 1 g/l 1-hexanesulphonicacid adjusted to pH 2.5 with H 3 PO 4 , B) acetonitrileGradient: 8 – 33% B in 30 min, then 5 min 90% BFlow rate: 0.32 ml/min, temperature: 40 °CDetection: DAD, 210 nmPeaks:1. Paracetamol2. Caffeine3. 6-Monoacetylmorphine4. Heroin5. Papaverine6 6. Noscapine4courtesy of Mr. Kothe,Laboratory Dr. Krone,Prof. Hagedorn,Medizinal-UntersuchungsstelleHerford35Analysis of customary ecstasyColumn: 250 x 4 mm NUCLEOSIL ® 100-5 C 18 HDEluents: A) 2 mmol NaH 2 PO 4 adjusted to pH 2.5with H 3 PO 4B) acetonitrile mAUGradient: 8– 33% B in 30 min230Flow rate: 0.8 ml/minDetection: UV, 235 nmPeak:1. Caffeine ?2. S(+)-3,4-Methylenedioxymethamphetaminehydrochloride (Ecstasy)courtesy of Mr. Kothe, 20Laboratory Dr. Krone, Prof. Hagedorn,Medizinal-Untersuchungsstelle Herford110130100110 20 30 min0 10 20 30min6.7.05MNwww.mn-net.com35

Columns for HPLCColumns for HPLCBase deactivated NUCLEOSIL ® phases for RP chromatographyApplicationsSeparation of tricyclic antidepressantsColumn: 250 x 4 mm NUCLEOSIL ® 100-5 Protect IEluent: acetonitrile – 25 mM KH 2 PO 4 , pH 7.0 (35 : 65, v/v)Flow rate: 1 ml/minTemperature: 25 °CDetection: UV, 254 nmPeaks:1. Protriptyline3. DoxepinO5. Amitriptyline1234562. NortriptylineNHCH 3N(CH 3 ) 24. ImipramineNNHCH 3N(CH 3 ) 26. TrimipramineNN(CH 3 ) 2N(CH 3 ) 21100300 5 10 15 minSeparation of tricyclic antidepressantsColumn: 250 x 4 mm NUCLEOSIL ®100-5 C 8 HD3Eluent: MeOH – 25 mM NaH 2 PO 4pH 7.0 (70:30)Flow rate 1 ml/minTemperature 30 °CDetection UV, 254 nmPeaks:141. Protriptyline2. Nortriptyline23. Doxepin4. Amitriptyline5. Trimipramine5110750Separation of tricyclic antidepressantsColumn: 250 x 4NUCLEOSIL ® 100-5 C 18 HDInjection volume 5 µlEluent acetonitrile – 0.1%TEAPO 4 pH 6.9 (45:55)Flow rate 1 ml/minTemperature 25 °CDetection UV, 254 nmPeaks:1. Protriptyline2. Nortriptyline3. Doxepin4. Imipramine5. Amitriptyline6. Trimipramine12 3 4 561107606.7.050 10 20 30min0 10 20min36www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Standard phases for RP chromatographyReversed phase silica phases are by far the most widelyused packings. Our base deactivated high performance RPmaterials are described in the preceding chapter. In thischapter you will find columns packed with standard silicaphases for RPLC. For ordering information of the correspondingbulk packings please see the chapter starting onpage 143. Octadecyl, Octyl, Butyl, Dimethyl and Phenylphases differ in retention time for a given solute. Under identicalconditions (mobile phase, flow rate, temperature etc.),e. g. the retention times on NUCLEOSIL ® C 18 are, for quite anumber of compounds, by a factor of up to 2 larger than onNUCLEOSIL ® C 8 .The mechanism of separation with all NUCLEOSIL ® andNUCLEODUR ® RP phases is the attraction between hydrophobicgroups of the test molecule and the bonded alkylgroups of the stationary phase by van der Waals forces. theless polar the sample molecules, the more they are attracted.Differences in the chemical structure of the sample moleculesresult in a stronger or weaker adsorption to the stationaryphase and hence in a separation.The more polar the sample, the shorter are the retentiontimes and the weaker are the van der Waals forces. The followingis roughly the order of increasing polarity (reverse orderof elution): aliphatic hydrocarbons, induced dipoles(CCI 4 ), permanent dipoles (CHCI 3 ), weak Lewis bases(ethers, aldehydes, ketones), strong Lewis bases (amines),weak Lewis acids (alcohols, phenols) and strong Lewis acids(carboxylic acids). Also of considerable importance for retentiontimes is the chain length of the molecules which are tobe separated.The mobile phase in RP chromatographyη[cP]at20 °C3viscosityViscosity of solvent mixturesas a function of composition3value. Higher concentrations of organic solvent will causeshorter retention times. RP separations can be optimised byutilising the specific selectivity of organic solvents.The viscosity of the eluent should not be neglected. It is oftennot realised in practice, that eluents have very different viscosities(η) and that these differences in viscosity (with thesame flow rate) cause changes in back pressure. The backpressure changes according to the following equation:ηp 11 = ----- ⋅ pη 2221watern-propanol2ethanolmethanolacetonitrile0 20 40 60 80 100% [w/w]organicThe mobile phase for reversed phase chromatography is amixture of water and an organic solvent which is misciblewith water. The time of elution of the substances to be separateddepends to a large degree on the water content of theorganic solvent and is therefore easily adjusted to a suitable1The viscosity of a solvent mixture is often higher than that ofthe single components, i.e. a higher pressure is needed toachieve the same flow rate.The graph on the left illustrates how strongly the viscosity ofthe eluent is influenced by its composition. For example theviscosity (and consequently the back pressure of the column)increases more than double when the eluent ischanged from acetonitrile – water (70:30) to methanol – water(70:30).When water is used as a solvent or as a component in gradientmixtures, it is essential that it is completely degassed.Apparent instability can be caused by air bubbles (from notproperly degassed solvents) which accumulate at the detector.The best way to remove such air is to first pump degassedsolvent through the column and then follow up withdegassed water. One can degas solvents with a vacuumpump, by passing helium through or by ultrasonic treatment.6.7.05MNwww.mn-net.com37

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Test mixture for reversed phase columnsFor reversed phase HPLC columns we supply a test mixturein acetonitrile to determine the quality of the packing. The figureon the right shows a typical test chromatogram of a columnwith the stationary phase NUCLEOSIL ® 100-5 C 18 .Ordering informationDesignation Pack of Cat. No.Test mixture for reversed phasecolumns in acetonitrile1 ml 722394Test mixture for reversed phase HPLC columns(Cat. No. 722394)Column: 250 x 4 mm NUCLEOSIL ® 100-5 C 18Sample volume: 20 µlEluent: acetonitrile – water (70:30)Flow rate: 1 ml/minPressure: 105 barTemperature: 22 °C2 3Detection: UV, 254 nm1Peaks: (concentrationin µg/ml eluent)1. Phenol (400)2. Naphthalene (180)3. Anthracene (12)4 8 min6.7.05Individual test certificate enclosed witheach NUCLEODUR ® and NUCLEOSIL ®column, respectively.38www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Octadecyl phases -(CH 2 ) 17 – CH 3Analysis of nitroaromatic compounds(explosives)Column 250 x 3 mm NUCLEOSIL ® 120-3 C 18 ; eluents: A) methanol,B) water, gradient 35 – 55% A in 12 min, then 40 min isocratic,finally 100% A, flow rate 0.35 ml/min; temperature 30 °C,detection DAD, 230 nm; injection volume 40 µlPeaks:1) 2,6-Diamino-4-nitrotoluene2) 2,4-Diamino-4-nitrotoluene3) Hexogene (Hexahydro-1,3,5-trinitro-1,3,5-triazine)4) 1,3,5-Trinitrobenzene5) 2-Methyl-3-nitroaniline6) 2-Methyl-5-nitroaniline7) 4-Methyl-3-nitroaniline200100mAU123456878) 1,3-Dinitrobenzene9) 2,4,6-Trinitrotoluene10)4-Amino-2,6-dinitrotoluene11)2-Amino-4,6-dinitrotoluene12)2,6-Dinitrotoluene13)2,4-Dinitrotoluene14)2-Nitrotoluene15)4-Nitrotoluene16)3-Nitrotoluene910 111312114810C 18 phases are suited for reversed phase chromatographyand ion-pairing chromatography for separation of nonpolar tomoderately polar compounds such asfatty acidsglyceridespolycyclic aromaticsesters (phthalates)fat-soluble vitaminssteroidsprostaglandinsPTH amino acids etc.Separation of triazines3 4 5Column: 250 x 4 mm NUCLEOSIL ® 50-5 C 18 ecEluent: methanol – acetonitrile – water (40:20:40, v/v/v)Flow rate: 1 ml/min, 30 °C, 220 barDetection: UV, 220 nmPeaks (injection volume 5 µl, 10 µg/ml each):1. Simazine 12. Atrazine3. Prometon 24. Ametryne5. Propazine6. Prometryne7. Terbutryne6 711157014 15 16020 40 mincourtesy of Dr. Steinbach, Inst. f. Org. Chemistry, Univ. Marburg010 20 minOrdering information · Analytical columns with NUCLEOSIL ® C 18Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 50-5 C 18 ecParticle size 5 µm, pore size 50 Å; octadecyl phase, endcapped, 14.5% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721825.20 721826.20 721828.20 721829.303 mm ID 721825.30 721826.30 721828.30 721829.304 mm ID 721825.40 721826.40 721828.40 721829.404.6 mm ID 721825.46 721826.46 721827.46 721828.46 721829.40EC columns 3)4 mm ID 720098.40 721829.404.6 mm ID 720098.46 721829.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other phases and other lengths are available on requestMNwww.mn-net.com39

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 18Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-3 C 18Particle size 3 µm, pore size 100 Å; octadecyl phase, endcapped, 15% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721883.20 721865.20 721866.303 mm ID 721883.30 721865.30 721866.304 mm ID 721883.40 721865.40 721866.404.6 mm ID 721883.46 721806.46 721865.46 721866.40Microbore columns 2)1 mm ID 717029.10 717020.10 717011.10 717003.10EC columns 3)2 mm ID 720150.20 720133.20 721866.303 mm ID 720150.30 720133.30 721866.304 mm ID 720150.40 720133.40 721866.404.6 mm ID 720150.46 720949.46 720133.46 721866.40NUCLEOSIL ® 100-5 C 18Particle size 5 µm, pore size 100 Å; octadecyl phase, endcapped, 15% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721074.20 721781.20 721622.20 721662.20 721602.303 mm ID 721074.30 721781.30 721622.30 721662.30 721602.304 mm ID 721074.40 721781.40 721622.40 721662.40 721602.404.6 mm ID 721074.46 721781.46 721622.46 721642.46 721662.46 721602.40Microbore columns 2)1 mm ID 717028.10 717019.10 717010.10 717002.10EC columns 3)2 mm ID 720002.20 720014.20 721602.303 mm ID 720002.30 720014.30 721602.304 mm ID 720002.40 720014.40 721602.404.6 mm ID 720002.46 720120.46 720014.46 721602.40NUCLEOSIL ® 100-7 C 18Particle size 7 µm, pore size 100 Å; octadecyl phase, endcapped, 15% C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721878.30 721609.304 mm ID 721878.40 721609.404.6 mm ID 721609.46EC columns 3)4 mm ID 720018.404.6 mm ID 720018.4630 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.0540Each column is individually tested and supplied with test chromatogram and test conditionswww.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 18Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-10 C 18Particle size 10 µm, pore size 100 Å; octadecyl phase, endcapped, 15% C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721681.30 721689.304 mm ID 721681.40 721689.404.6 mm ID 721689.46EC columns 3)4 mm ID 720023.404.6 mm ID 720023.46NUCLEOSIL ® 120-3 C 18Particle size 3 µm, pore size 120 Å; octadecyl phase, endcapped, 11% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721075.20 721721.20 721626.20 721666.20 721606.303 mm ID 721075.30 721721.30 721626.30 721666.30 721606.304 mm ID 721075.40 721721.40 721626.40 721666.40 721606.404.6 mm ID 721075.46 721721.46 721626.46 721646.46 721666.46 721606.40Microbore columns 2)1 mm ID 717031.10 717022.10 717013.10 717005.10EC columns 3)2 mm ID 720040.20 720055.20 721606.303 mm ID 720040.30 720055.30 721606.304 mm ID 720040.40 720055.40 721606.404.6 mm ID 720040.46 720740.46 720055.46 721606.40NUCLEOSIL ® 120-5 C 18Particle size 5 µm, pore size 120 Å; octadecyl phase, endcapped, 11% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721070.20 721723.20 721629.20 721712.20 721783.303 mm ID 721070.30 721723.30 721629.30 721712.30 721783.304 mm ID 721070.40 721723.40 721629.40 721712.40 721783.404.6 mm ID 721070.46 721723.46 721629.46 721659.46 721712.46 721783.40Microbore columns 2)1 mm ID 717030.10 717021.10 717012.10 717004.10EC columns 3)2 mm ID 720051.20 720041.20 721783.303 mm ID 720051.30 720041.30 721783.304 mm ID 720051.40 720041.40 721783.404.6 mm ID 720051.46 720730.46 720041.46 721783.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05MNCustom-packed columns with other phases and other lengths are available on requestwww.mn-net.com41

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 18Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columns6.7.05NUCLEOSIL ® 120-7 C 18Particle size 7 µm, pore size 120 Å; octadecyl phase, endcapped, 11% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720042.404.6 mm ID 720042.46NUCLEOSIL ® 120-10 C 18Particle size 10 µm, pore size 120 Å; octadecyl phase, endcapped, 11% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720043.404.6 mm ID 720043.46NUCLEOSIL ® 300-5 C 18Particle size 5 µm, pore size 300 Å; octadecyl phase, endcapped, 6.5% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721790.20 721628.20 721668.20 721608.303 mm ID 721628.30 721668.30 721608.304 mm ID 721628.40 721668.40 721608.404.6 mm ID 721628.46 721648.46 721668.46 721608.40Microbore columns 2)1 mm ID 717045.10 717048.10 717056.10 717059.10EC columns 3)2 mm ID 720713.20 721608.303 mm ID 720713.30 721608.304 mm ID 720065.40 721608.404.6 mm ID 720065.46 721608.40NUCLEOSIL ® 500-7 C 18Particle size 7 µm, pore size 500 Å; octadecyl phase, endcapped, 2% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720074.404.6 mm ID 720074.46NUCLEOSIL ® 1000-7 C 18Particle size 7 µm, pore size 1000 Å; octadecyl phase, endcapped, ~1% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720077.404.6 mm ID 720077.46NUCLEOSIL ® 4000-7 C 18Particle size 7 µm, pore size 4000 Å; octadecyl phase, endcapped,

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Wide pore silica packingsSilica as HPLC packing material has several advantages:pore size and surface are definedit is mechanically stablenumerous methods for surface modification areavailableMany biologically interesting molecules can not be separatedusing conventional narrow pore silicas with pore sizes of about100 Å. This is why we offer a complete line of wide pore packingswith pore sizes of 300, 500, 1000 and 4000 Å. These materialscan also be used for size exclusion chromatography(SEC).Octyl phases -(CH 2 ) 7 – CH 3C 8 phases are suited for reversed phase chromatographyand ion-pairing chromatography for separation of moderatelyto highly polar (water-soluble) compounds such assteroidsnucleosidescyclodextrinspharmacological plant constituentsAnthranoids in senna based drugsL. Kabelitz, K. Reif, Dtsch. Apotheker Ztg. 134 (1994) 37 – 40Column: 250 x 4 mm NUCLEOSIL ® 100-5 C 8Eluents: A) 0.02 M KH 2 PO 4 , adjusted to pH 2 with H 3 PO 4 (85%), B) acetonitrileGradient: flow rate 1 ml/min: 10 min 14% B, in 5 min to 20% B, 10 min at 20% B, flow rate 0.8 ml/min: in 1 min to 21% B, in 29 min to 80% B,flow rate 1 ml/min: in 2 min to 14% B, then 13 min at 14% BPhotodiode array: dianthrones at 270 nm, anthraquinones at 435 nm12Detection:Peaks:1. Aloe emodin-8-glucoside2. Rhein-8-glucoside3: Sennoside D 14: Sennoside D5: Sennoside B6: Sennoside A 17: Sennoside C8: Sennoside A9: Sennidin-B-monoglucoside10: Sennidin-A-monoglucoside11: Aloe emodin12: Rhein13: Sennidin A14: Emodin15: Sennidin B0.60.50.40.30.20.1123 4 56789 10 1113141510263000 10 20 30 40 minOrdering information · Analytical columns with NUCLEOSIL ® C 8Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 50-5 C 8 ecParticle size 5 µm, pore size 50 Å; octyl phase, endcapped, 9% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721830.20 721831.20 721833.20 721834.303 mm ID 721830.30 721831.30 721833.30 721834.304 mm ID 721830.40 721831.40 721833.40 721834.404.6 mm ID 721830.46 721831.46 721832.46 721833.46 721834.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) 30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2) On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other phases and other lengths are available on requestMNwww.mn-net.com43

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 8Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsEC columns 3)4 mm ID 720092.40 721834.404.6 mm ID 720092.46 721834.40NUCLEOSIL ® 100-5 C 8 ecParticle size 5 µm, pore size 100 Å; octyl phase, endcapped, 9% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721804.20 721795.20 721796.20 721805.303 mm ID 721804.30 721795.30 721796.30 721805.304 mm ID 721804.40 721795.40 721796.40 721805.404.6 mm ID 721804.46 721795.46 721797.46 721796.46 721805.40Microbore columns 2)1 mm ID 717035.10 717026.10 717017.10 717008.10NUCLEOSIL ® 100-5 C 8Particle size 5 µm, pore size 100 Å; octyl phase, not endcapped, 8.5% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721194.20 721780.20 721621.20 721661.20 721601.303 mm ID 721194.30 721780.30 721621.30 721661.30 721601.304 mm ID 721194.40 721780.40 721621.40 721661.40 721601.404.6 mm ID 721194.46 721780.46 721621.46 721641.46 721661.46 721601.40Microbore columns 2)1 mm ID 717036.10 717027.10 717018.10 717009.10EC columns 3)3 mm ID 720001.30 720013.30 721601.304 mm ID 720001.40 720013.40 721601.404.6 mm ID 720001.46 720990.46 720013.46 721601.40NUCLEOSIL ® 100-7 C 8Particle size 7 µm, pore size 100 Å; octyl phase, not endcapped, 8.5% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720017.404.6 mm ID 720017.46NUCLEOSIL ® 100-10 C 8Particle size 10 µm, pore size 100 Å; octyl phase, not endcapped, 8.5% C; eluent in column acetonitrile / waterEC columns 3)4 mm ID 720022.404.6 mm ID 720022.4630 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.0544Each column is individually tested and supplied with test chromatogram and test conditionswww.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 8Length → 30 mm 1) 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 120-3 C 8Particle size 3 µm, pore size 120 Å; octyl phase, not endcapped, 6.5% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721720.20 721786.20 721782.20 721785.303 mm ID 721720.30 721786.30 721782.30 721785.304 mm ID 721720.40 721786.40 721782.40 721785.404.6 mm ID 721720.46 721786.46 721722.46 721782.46 721785.40EC columns 3)2 mm ID 720071.20 720703.20 721785.303 mm ID 720071.30 720703.30 721785.304 mm ID 720071.40 720703.40 721785.404.6 mm ID 720071.46 720214.46 720703.46 721785.40NUCLEOSIL ® 120-5 C 8Particle size 5 µm, pore size 120 Å; octyl phase, not endcapped, 6.5% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721868.20 721892.20 721801.20 721787.303 mm ID 721868.30 721892.30 721801.30 721787.304 mm ID 721868.40 721892.40 721801.40 721787.404.6 mm ID 721868.46 721892.46 721521.46 721801.46 721787.40EC columns 3)2 mm ID 720050.20 720052.20 721787.303 mm ID 720050.30 720052.30 721787.304 mm ID 720050.40 720052.40 721787.404.6 mm ID 720050.46 720735.46 720052.46 721787.40NUCLEOSIL ® 300-5 C 8Particle size 5 µm, pore size 300 Å; octyl phase, not endcapped, ~3% C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721103.30 721098.30 721101.304 mm ID 721103.40 721098.40 721101.40EC columns 3)4 mm ID 720062.40 721101.404.6 mm ID 720062.46 721101.4030 mm ChromCart ® columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)30 mm ChromCart ® columns require the CC column holder 30 mm (Cat. No. 721823).2)On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.3)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).www.mn-net.com6.7.05MNCustom-packed columns with other phases and other lengths are available on requestwww.mn-net.com45

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Phenyl phases -(CH 2 ) 3C 6 H 5 phases are suited for reversed phase chromatographyand ion pairing chromatography for separation of moderatelypolar compounds. The retention characteristics are similar tothe C 8 phase, but with different selectivity for polycyclic aromatichydrocarbons, polar aromatics, fatty acids etc.Separation of opium alkaloids and meconic acidN.R. Ayyangar, S. R. Bhide, J. Chromatogr. 436 (1988) 455-465Column: 250 x 4 mm NUCLEOSIL ® 100-7 C 6 H 5Eluent A: methanol – water (5:95) with 1% triethylammoniumphosphate stock buffer, pH 3.2Eluent B: methanol – water (80:30) with 1% triethylammoniumphosphate stock buffer, pH 3.95Gradient: linear from 100% A to 100% B in 20 min,then 12 min eluent BFlow rate: 1,5 ml/minDetection: UV, 280 nmPeaks:1. Morphine2. Meconic acid3. Codeine4. Thebaine5. Laudanosine6. Cryptopine7. Noscapine8. Narceine9. Papaverine% v CH 3 OH10090807060504030201001230 2 4 6 8 10 12 14 16 18 20 22 24 26min45 6 7891012006.7.05Ordering information · Analytical columns with NUCLEOSIL ® C 6 H 5Length → 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 C 6 H 5 ecParticle size 5 µm, pore size 100 Å; phenyl phase, endcapped, 8 % C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721535.30 721533.30 721537.304 mm ID 721535.40 721533.40 721537.40NUCLEOSIL ® 100-5 C 6 H 5Particle size 5 µm, pore size 100 Å; phenyl phase, not endcapped, 8% C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721860.30 721861.30 721862.304 mm ID 721860.40 721861.40 721862.404.6 mm ID 721860.46 721887.46 721861.46 721862.40EC columns 1)2 mm ID 720695.20 721862.303 mm ID 720695.30 720956.30 721862.304 mm ID 720956.40 721862.404.6 mm ID 720956.46 721862.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Each column is individually tested and supplied with test chromatogram and test conditions46www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 6 H 5Length → 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-7 C 6 H 5Particle size 7 µm, pore size 100 Å; phenyl phase, not endcapped, 8% C; eluent in column acetonitrile / waterEC columns 1)4 mm ID 720019.404.6 mm ID 720019.46ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Butyl phases -(CH 2 ) 3 – CH 3C 4 phases are suited for reversed phase chromatographyand ion-pairing chromatography for separation of macromoleculesand hydrophobic substances. Retention times onHemoglobin analysis of the adult Sumatran tigerbutyl phases are shorter than on C 8 and C 18 phases. Forbutyl phases for biochemical separation problems please referto page 107.M. Jahan et al. Biol. Chem. Hoppe-Seyler 370 (1989) 27 – 33Column: 250 x 4.6 mm NUCLEOSIL ® 300-5 C 4Eluents: A) 0.1% trifluoroacetic acid, B) acetonitrileGradient: 0 – 35% B in 2 min, then 35 – 60% B in 60 minFlow rate: 1 ml/minDetection: UV, 230 nmA 230HemeαβIIβI1135000 10 20 30 40 50 60 minOrdering information · Analytical columns with NUCLEOSIL ® C 4Length → 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 120-5 C 4Particle size 5 µm, pore size 120 Å; butyl phase, endcapped, ~2 % C; eluent in column acetonitrile / waterChromCart ® columns3 mm ID 721891.30 721890.30 721889.304 mm ID 721891.40 721890.40 721889.40EC columns 2)4 mm ID 720096.40 721889.404.6 mm ID 720096.46 721889.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.2)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05MNCustom-packed columns with other phases and other lengths are available on requestwww.mn-net.com47

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Ordering information · Analytical columns with NUCLEOSIL ® C 4Length → 70 mm 100 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 300-5 C 4Particle size 5 µm, pore size 300 Å; butyl phase, endcapped, ~2% C; eluent in column acetonitrile / waterChromCart ® columns2 mm ID 721784.20 721627.20 721667.20 721607.303 mm ID 721627.30 721667.30 721607.304 mm ID 721627.40 721667.40 721607.404.6 mm ID 721627.46 721647.46 721667.46 721607.40Microbore columns 1)1 mm ID 717044.10 717047.10 717055.10 717058.10EC columns 2)4 mm ID 720059.40 721607.404.6 mm ID 720059.46 721607.40NUCLEOSIL ® 300-7 C 4Particle size 7 µm, pore size 300 Å; butyl phase, endcapped, ~2% C; eluent in column acetonitrile / waterEC columns 2)4 mm ID 720060.404.6 mm ID 720060.46ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.Guard columns for Microbore columns on request.2) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Dimethyl phases -(CH 3 ) 2Dimethyl phases are suited for reversed phase chromatographyand ion-pairing chromatography. Retention times on thedimethyl phase are much shorter than for the other RP phases.Ordering information · Analytical columns with NUCLEOSIL ® C 2Length → 125 mm 250 mm Guard columnsNUCLEOSIL ® 100-7 C 2Particle size 7 µm, pore size 100 Å; dimethyl phase, not endcapped, 3.5 % C; eluent in column acetonitrile / waterChromCart ® columns4 mm ID 721873.40 721874.40 721069.40EC columns 1)4 mm ID 720089.40 721069.404.6 mm ID 720089.46 721069.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Each column is individually tested and supplied with test chromatogram and test conditions48www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Unmodified NUCLEOSIL ® , polar NUCLEOSIL ® phases and ion exchangersPackings with chemically bonded polar phasesThese materials are characterised by fast mass transfer, becausethe bonded organic layer of these stationary phases ismonomeric, thus minimising diffusion paths.Due to the availability of different functional groups it is possibleto vary the chemical characteristics of the surface andconsequently the adsorption characteristics of the stationaryphase. Silica packings with bonded polar phases thus combinethe advantages of partition chromatography (variation ofadsorption properties) and adsorption chromatography (rapidmass transfer in the stationary phase). Further advantagesof chemically bonded polar phases are:They do not “bleed”; equilibration with a liquid stationaryphase is not necessary as is the case in classical partitionchromatography.The influence of impurities in the eluent (e. g. water) onthe separation qualities of bonded polar phases is muchsmaller than is the case with unmodified silica gels andaluminium oxides for adsorption chromatography.Swelling and consequently variation in the permeabilityof the packed columns never occurs with chemicallybonded polar silica phases. Even the use of water orother aqueous eluents does not present any problems.Although the chemically bonded phases on the surface orthe silica gel form a monomeric organic layer, they have alarge surface concentration of functional groups, which isabout 3 to 4 functional groups per 100 Å 2 , i. e. about5 µeq/m 2 .Influence of functional groups on the separation characteristics of a stationary phasek’10,09,08,07,06,0Effect of modification on the separationof aromatic hydrocarbonsNO 2 phaseThe figure on the left shows the influence of the modificationof silica gel on the k’ values (capacity factors) of aromatic hydrocarbons,with n-heptane as eluent on unmodified NUCL-EOSIL ® as well as three chemically bonded polar phases,namely NUCLEOSIL ® CN, NH 2 and NO 2 . On the y-axis thek’ values of 9 aromatic hydrocarbons have been plotted. Theinfluence of the nitroaromatic groups on the separation is especiallyevident. A strong interaction between the electronicsystems results in the formation of charge transfer complexeswith considerably larger k’ values than for unmodifiedNUCLEOSIL ® .5,0NH 2 phase4,03,02,0NUCLEOSILunmodified1,00BenzeneNaphthaleneBiphenylAnthracenePhenanthrenePyreneChryseneBenzo[a]pyreneCN phaseBenzo[e]pyrene6.7.05MNwww.mn-net.com49

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®NUCLEOSIL ® ion exchangersThese phases have been especially developed for HPLC.Contrary to adsorptions or reversed phase chromatographywith ion exchange chromatography there are three variableparameters for the mobile phase, which can influence theseparationthe type of buffer,the ionic strength andthe pH value.By changing the ionic strength of the eluent, the competitionbetween mobile phase and sample molecules for the functionalionic groups on the stationary phase is influenced.When the retention time of the sample molecules is too long,an increase in the ionic strength of the mobile phase will reduceretention times and vice versa.A change in the pH value of the mobile phase will alter thedissociation of analyte and buffer salt. These two effects influencethe equilibrium between the sample molecules andthe functional groups of the stationary phase. Thus the retentionof weak acids on an anion exchanger will increase whenthe pH value is raised, because a larger part of the acid ispresent in ionic form at higher pH values. If, on the otherhand, e.g. a phthalate buffer is used, increase of the pH valuecan increase the concentration of phthalate ions and thusthe ionic strength. This results in a decreasing retention time.Proper selection of buffer types and pH values can thus resultin enormous selectivity differences, which can be utilisedfor optimisation of a separation.Even for equal ionic strength of the mobile phase, differentions show different degrees of affinity to the ion exchangegroups. The following orders of increasing affinity can be givenfor anion exchangersF – < OH – < Cl – < CN – < Br – < NO – 3 < HSO – 4 < I –and for cation exchangersH + < Na + < NH + 4 < K + < Cs + < Ag +Divalent ions are more strongly attracted to the ion exchangegroups than monovalent, and trivalent more than divalentions. Generally speaking, the greater the charge and effectiveradius of ions, the stronger is the attraction to the ion exchangegroups.+ + ++ +++ + +++ ++++ + + +++++ ++ ++++ +++www.mn-net.com6.7.0550www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Cyano phases (nitrile)Trace analysis of tetracyclinesC. Fu, H. Xu, Proc. 6th Natl. Symp.Chromatogr. (1987) 412 – 4131 2 3Column250 x 4.6 mm NUCLEOSIL ® 100-5 CNEluent acetonitrile – 0.015 M oxalic acidpH 2.35 (20:80); flow rate 1 ml/minDetection UV, 363 nm4 5 6Peaks:1. Terramycin2. E-Tetracycline3. Tetracycline4. E-Aureomycin5. Aureomycin6. Deoxyterramycin0 4 8 12 minOrdering information · Analytical columns with NUCLEOSIL ® CN103760Custom-packed columns with other phases and other lengths are available on request-(CH 2 ) 3 – CNCN phases are suited for reversed phase chromatographyand for normal phase chromatography:Normal phase chromatography:with low-polarity solvents for many compounds, which can alsobe separated on unmodified silica, however, due to therapid equilibration much more suitable for gradient separationsReversed phase chromatography:with different selectivity than C 18 , C 8 or phenyl modified packingsPlease note: the eluent in the columns (except withNUCLEOSIL ® 100-5 CN-RP) is n-heptane. If you want to usean eluent which is not miscible with n-heptane (e. g. water), itis necessary to rinse the column with THF first.Length → 70 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 CNParticle size 5 µm, pore size 100 Å; cyano phase (nitrile), eluent in column n-heptaneChromCart ® columns2 mm ID 721788.20 721624.20 721664.20 721604.303 mm ID 721624.30 721664.30 721604.304 mm ID 721624.40 721664.40 721604.404.6 mm ID 721624.46 721644.46 721664.46 721604.40EC columns 1)4 mm ID 720090.40 721604.404.6 mm ID 720090.46 721604.40NUCLEOSIL ® 100-5 CN-RPParticle size 5 µm, pore size 100 Å; cyano phase (nitrile), eluent in column acetonitrile / waterEC columns 1)4 mm ID 720205.40 721917.404.6 mm ID 720205.46 721917.40NUCLEOSIL ® 100-10 CNParticle size 10 µm, pore size 100 Å; cyano phase (nitrile), eluent in column n-heptaneEC columns 1)4 mm ID 720024.404.6 mm ID 720024.46NUCLEOSIL ® 120-7 CNParticle size 7 µm, pore size 120 Å; cyano phase (nitrile), eluent in column n-heptaneEC columns 1)4 mm ID 720057.404.6 mm ID 720057.46ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05MNwww.mn-net.com51

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Cyano phases · ApplicationLC/MS of the oxidation products of cholesterolM. Careri et al. J. Chromatogr. 794 (1998) 253 – 262Column: 250 x 2 mm NUCLEOSIL ® 100-5 CNEluent: heptane – propan-2-ol (94:6)Flow rate: 0.3 ml/minDetection: MS; (a) EI mode, (b) PCI mode, (c) NCI modeElution sequence: 1 cholesterol, 2 5,6α-epoxycholesterol, 3 25-hydroxycholesterol, 4 7-ketocholesterol, 5 7β-hydroxycholesterol,6 Cholestan-3β-5α-6β-triolNitro phases · ApplicationSeparation of gibberellins by normal phase HPLCP.C. Odén et al. J. Chromatogr. 464 (1989) 195 – 200Column: 125 x 4.6 mm NUCLEOSIL ® 100-5 NO 2Eluent A: n-heptane half saturated with 1 M formic acidEluent B: ethyl acetate with 1% water and 0.5% formic acidGradient: 100% A – 100% B in 60 minFlow rate: 2 ml/minDetection: radioactivity100% = 783151005000 2 4 6 8 10 12 14 16 min100% = 12136(b)1005000 2 4 6 8 10 12 14 16 min(a)Radioactivitykaurene kaurenalkaurenolkaurenoic acidGA 12 -aldGA 9GA 15GA12GA 4GA 20GA 44GA 53GA 1GA 3GA 19GA 29GA 8100% = 19350100(c)1120305000 2 4 6 8 10 12 14 16 min10628010 20 30 40 50 60 70 80 90 100 110 120Elution volume(ml)Nitro phases -(CH 2 ) 3 NO 2NO 2 phases are suited for the separation of compounds withdouble bonds or for aromatic compounds.Please note: the eluent in the columns is n-heptane. If youwant to use an eluent which is not miscible with n-heptane(e. g. water), it is necessary to rinse the column with THFfirst.Ordering information · Analytical columns with NUCLEOSIL ® NO 2Length → 125 mm 250 mm Guard columns6.7.05NUCLEOSIL ® 100-5 NO 2Particle size 5 µm, pore size 100 Å; nItro phase, eluent in column n-heptaneChromCart ® columns3 mm ID 721864.30 721707.30 721863.304 mm ID 721864.40 721707.40 721863.40EC columns 1)4 mm ID 720993.40 721863.404.6 mm ID 720993.46 721863.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Each column is individually tested and supplied with test chromatogram and test conditions52www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Diol phasesSFC separation of steroidsF. Vérillon et al. LC · GC int. 7 (1994) 710 – 716Column: 250 x 2 mm NUCLEOSIL ® 100-7 OH (Diol)Eluent: CO 2 with 1 – 12% methanolFlow rate: 2.5 ml/minOutlet pressure: 14 – 35 MpaETemperature: 85 °CDetection: UV, 210 nmPeaks:KA) CholesterolB) ProgesteroneGC) AndrosteroneA FD) TestosteroneHI JB DE) EstroneF) EstradiolCG) CortisoneH) HydrocortisoneI) PrednisoneJ) PrednisoloneK) Estriol109840-(CH 2 ) 3 – O – CH 2 – CH – CH 2–OH–OHOH phases are suited for reversed phase chromatographyand normal phase chromatography; they are less polar thanunmodified silica and very easily wettable with water.Please note: the eluent in the columns is n-heptane. If youwant to use an eluent which is not miscible with n-heptane(e. g. water), it is necessary to rinse the column with THFfirst.0 2 4 min 6The hollow in the upper trace is caused by instantaneous detection ofthe pressure gradient and delayed detection of the methanol gradient.The lower trace shows a software-corrected chromatogram.Ordering information · Analytical columns with NUCLEOSIL ® OH (Diol)Length → 125 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 OH (Diol)Particle size 5 µm, pore size 100 Å; diol phase, eluent in column n-heptaneChromCart ® columns3 mm ID 721480.30 721471.30 721478.304 mm ID 721480.40 721471.40 721478.40EC columns 1)4 mm ID 720143.40 721478.404.6 mm ID 720143.46 721478.40NUCLEOSIL ® 100-7 OH (Diol)Particle size 7 µm, pore size 100 Å; diol phase, eluent in column n-heptaneEC columns 1)4 mm ID 720070.404.6 mm ID 720070.46ChromCart ® guard columns cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other phases and other lengths are available on requestMNwww.mn-net.com53

Columns for HPLCColumns for HPLC6.7.05Analytical columns packed with NUCLEOSIL ®Unmodified silicaDue to its narrow pore structure, NUCLEOSIL ® 50 is recommendedfor adsorption chromatography, while NUCLEOSIL ®100 can be used for adsorption as well as for partition chromatography.SiOHPlease note: the eluent in the columns is n-heptane. If youwant to use an eluent which is not miscible with n-heptane(e. g. water), it is necessary to rinse the columns with THFfirst.Ordering information · Analytical columns with unmodified NUCLEOSIL ®Length → 70 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 50-5Particle size 5 µm, pore size 50 Å; unmodified, eluent in column n-heptaneChromCart ® columns2 mm ID 721794.20 721620.20 721660.20 721600.303 mm ID 721620.30 721660.30 721600.304 mm ID 721620.40 721660.40 721600.404.6 mm ID 721620.46 721640.46 721660.46 721600.40EC columns 1)4 mm ID 720093.40 721600.404.6 mm ID 720093.46 721600.40NUCLEOSIL ® 50-7Particle size 7 µm, pore size 50 Å; unmodified, eluent in column n-heptaneEC columns 1)4 mm ID 720015.404.6 mm ID 720015.46NUCLEOSIL ® 100-5Particle size 5 µm, pore size 100 Å; unmodified, eluent in column n-heptaneChromCart ® columns2 mm ID 721871.20 721870.20 721872.303 mm ID 721871.30 721870.30 721872.304 mm ID 721871.40 721870.40 721872.404.6 mm ID 721871.46 721516.46 721870.46 721872.40EC columns 1)3 mm ID 720099.30 721872.304 mm ID 720099.40 721872.404.6 mm ID 720099.46 721872.40NUCLEOSIL ® 100-7Particle size 7 µm, pore size 100 Å; unmodified, eluent in column n-heptaneEC columns 1)4 mm ID 720016.404.6 mm ID 720016.46NUCLEOSIL ® 100-10Particle size 10 µm, pore size 100 Å; unmodified, eluent in column n-heptaneEC columns 1)4 mm ID 720021.404.6 mm ID 720021.46ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1) As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Each column is individually tested and supplied with test chromatogram and test conditions54www.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Amino phases -(CH 2 ) 3 – NH 2NH 2 phases feature versatile applicability:– in normal phase chromatography with hexane, CH 2 CI 2 orisopropanol as mobile phase for polar compounds suchas substituted anilines, esters, chlorinated pesticides etc.– in aqueous-organic eluent systems for reversed phasechromatography of polar compounds like carbohydrates– as weak anion exchanger for anions and organic acidsusing common buffers (e. g. acetate or phosphate) inconjunction with organic modifiers (e. g. acetonitrile)Please note: the eluent in the columns is n-heptane. If youwant to use an eluent which is not miscible with n-heptane(e. g. water), it is necessary to rinse the column with THFfirst.NUCLEOSIL ® 100-5 NH 2 RPOrdering information · Analytical columns with NUCLEOSIL ® NH 2– the amino phase for reversed phase applications; eluentin column: acetonitrile / water– rinsing with THF is not requiredLength → 70 mm 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-3 NH 2Particle size 3 µm, pore size 100 Å; amino phase, eluent in column n-heptaneChromCart ® columns3 mm ID 721121.30 721120.30 721122.304 mm ID 721121.40 721120.40 721122.404.6 mm ID 721121.46 721120.46 721122.40EC columns 1)4 mm ID 720275.40 721122.404.6 mm ID 720275.46 721122.40NUCLEOSIL ® 100-5 NH 2Particle size 5 µm, pore size 100 Å; amino phase, eluent in column n-heptaneChromCart ® columns2 mm ID 721789.20 721625.20 721665.20 721605.303 mm ID 721625.30 721665.30 721605.304 mm ID 721625.40 721665.40 721605.404.6 mm ID 721625.46 721645.46 721665.46 721605.40EC columns 1)4 mm ID 720095.40 721605.404.6 mm ID 720095.46 721605.40NUCLEOSIL ® 100-5 NH 2 RPParticle size 5 µm, pore size 100 Å; amino phase, eluent in column acetonitrile / water (80:20)ChromCart ® columns3 mm ID 721625.30RP 721665.30RP 721605.30RP4 mm ID 721665.40RP 721605.40RP4.6 mm ID 721645.46 RP 721605.40RPEC columns 1)3 mm ID 720121.30RP 720095.30RP 721605.30RPNUCLEOSIL ® 120-7 NH 2Particle size 7 µm, pore size 120 Å; amino phase, eluent in column n-heptaneEC columns 1)4 mm ID 720058.404.6 mm ID 720058.46ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05MNCustom-packed columns with other phases and other lengths are available on requestwww.mn-net.com55

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Dimethylamino phases -(CH 2 ) 3 – N(CH 3 ) 2The DMA phase is a weakly basic anion exchanger for theseparation of many anions; it can also be used in a similarway as the NH 2 phase.Please note: the eluent in the columns is n-heptane. If youwant to use an eluent which is not miscible with n-heptane(e. g. water), it is necessary to rinse the column with THFfirst.Separation of sennosides from rhubarbY. Ohshima, K. Takahashi, J. Chromatogr. 258 (1983) 292 – 296Column: NUCLEOSIL ® 100-5 N(CH 3 ) 2 , 250 x 8 mm IDEluent: THF – water – acetic acid (8:2:1)Flow rate: 3,5 ml/minDetection: UV, 280 nmSamples: methanolic extracts ofa) Rheum palmatum var. tanguticumb) Rheum coreanumc) Rheum palmatumd) Rheum coreanum x Rheum palmatum (Shinshu-rhubarb)(d)(c)(b)(a)min 10 0 min 10 0 min 10 0 min 10 06.7.05102590Ordering information · Analytical columns with NUCLEOSIL ® N(CH 3 ) 2Length → 250 mm Guard columnsNUCLEOSIL ® 100-5 N(CH 3 ) 2Particle size 5 ± 1.5 µm, pore size 100 Å; dimethylamino phase, eluent in column n-heptaneEC columns4 mm ID 720994.40 721610.404.6 mm ID 720994.46 721610.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).56Each column is individually tested and supplied with test chromatogram and test conditionswww.mn-net.comMN

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Cation exchangers, strongly acidic -(CH 2 ) 3 SO 3 NaThe cation exchanger NUCLEOSIL ® SA is used for ion exchangechromatography.It is a strongly acidic cation exchanger with sulphonic acidmodification and a capacity of about 1 meq/g.Separation of rare-earth elementsNuryono et al. Chromatographia 48 (1998) 407 – 414Column: 150 x 4.6 mm NUCLEOSIL ® 100-5 SAEluent A: 20 mM α-hydroxyisobutyric acid (HIBA), pH 4.6Eluent B: 12 mM oxalic acid, pH 4.6Gradient: 8 min 5% B, 5 – 50% B in 10 min, 50 – 75% B in 2 min, 5 min 75%BFlow rate: 1 ml/minTemperature: 25 °CDetection: post-column derivatization with Arsenazo I, measurement in visiblelight at 600 nmSample: rare-earth elements, 10 ppm each, 20 µl injected806040mAU600 nmLuYbTmYGdSm Nd PrEuCeLaHoEr DyTb2011403000 5 10 15 20 minOrdering information · Analytical columns with NUCLEOSIL ® SALength → 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 SAParticle size 5 µm, pore size 100 Å; cation exchanger, strongly acidic, eluent in column 0.15 M (NH 4 ) 2 HPO 4 , pH 5ChromCart ® columns3 mm ID 721486.30 721342.30 721487.304 mm ID 721486.40 721342.40 721487.404.6 mm ID 721486.46 721525.46 721342.46 721487.40EC columns 1)4 mm ID 720097.40 721487.404.6 mm ID 720097.46 721487.40NUCLEOSIL ® 100-10 SAParticle size 10 µm, pore size 100 Å; cation exchanger, strongly acidic, eluent in column 0.15 M (NH 4 ) 2 HPO 4 , pH 5ChromCart ® columns3 mm ID 721881.30 721683.30 721706.304 mm ID 721881.40 721683.40 721706.40EC columns 1)4 mm ID 720028.40 721706.404.6 mm ID 720028.46 721706.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).6.7.05Custom-packed columns with other phases and other lengths are available on requestMNwww.mn-net.com57

Columns for HPLCColumns for HPLCAnalytical columns packed with NUCLEOSIL ®Anion exchangers, strongly basic -(CH 2 ) 3 CH 2 – N + (CH 3 ) 3 Cl –The anion exchanger NUCLEOSIL ® SB is used for ion exchangechromatography.It is a strongly basic anion exchanger with a quaternary ammoniummodification and a capacity of about 1 meq/g.www.mn-net.comHPLC of inorganic and organic anionsD. Yan, G. Schwedt, J. Chromatogr. 516 (1990) 383 – 393Column 250 x 4 mm NUCLEOSIL ® 100-10 SBEluent 1 mMdiethylenetriaminepentaaceticacid(DTPA)Flow rate2 ml/minDetectionconductivityAcetateCl – NO 2–FormateBr – NO 3–SO 2–4TartrateOxalate0.2 µS1065400 8 16 min6.7.05Ordering information · Analytical columns with NUCLEOSIL ® SBLength → 125 mm 150 mm 250 mm Guard columnsNUCLEOSIL ® 100-5 SBParticle size 5 µm, pore size 100 Å; anion exchanger, strongly basic, eluent in column 0.15 M (NH 4 ) 2 HPO 4 , pH 5ChromCart ® columns3 mm ID 721688.30 721884.30 721885.304 mm ID 721688.40 721884.40 721885.404.6 mm ID 721688.46 721523.46 721884.46 721885.40EC columns 1)4 mm ID 720996.40 721885.404.6 mm ID 720996.46 721885.40NUCLEOSIL ® 100-10 SBParticle size 10 µm, pore size 100 Å; anion exchanger, strongly basic, eluent in column 0.15 M (NH 4 ) 2 HPO 4 , pH 5ChromCart ® columns3 mm ID 721882.30 721879.30 721886.304 mm ID 721882.40 721879.40 721886.40EC columns 1)4 mm ID 720029.40 721886.404.6 mm ID 720029.46 721886.40ChromCart ® guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.1)As guard columns for EC columns use ChromCart ® guard column cartridges with guard column adaptor EC (Cat. No. 721359).Each column is individually tested and supplied with test chromatogram and test conditions58www.mn-net.comMN