Columns for HPLC - Western Analytical

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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
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Columns for HPLC - Western Analytical

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