NUCLEOSHELL core-shell silica for HPLC

NUCLESHELL core-shell silica for HPLC Core-shell technology µm 0. µm.8 µm 0. µm.7 µm 0. µm.7 µm 0. µm Key feature Solid core of silicon dioxide, homogeneous shell of porous silica Highest efficiency compared to traditional totally porous materials Pore size 90 Å; particle size.7 μm (core.7 μm) and μm (core.8 μm); specific surface 0 (.7 μm) and 90 ( μm) m /g lower back pressure enables use on conventional LC systems Pressure stability 00 bar Demands on HPLC separations are constantly increasing with respect to separation efficiency, detection limits, and the time requirements for each analysis. Several approaches have been made to achieve fast separations without losing chromatographic performance. HPLC columns packed with particles < μm show very high efficiencies (plates/meter) and allow the use of smaller column sizes with the positive side effect of significant solvent saving. However they generate a high back pressure of the mobile phase during column runs which requires specifically designed equipment. Columns: R s = N α α k' i k' i + R s = resolution, α = selectivity (separation factor), k i = retention N = plate number with N /d P, d P = particle diameter Resolution R S as function of particle size 0 x mm MN Appl. No. 70 NUCLESHELL RP 8,.7 μm NUCLEDUR C 8 Gravity, μm NUCLEDUR C 8 Gravity,.8 μm acetonitrile water (0:0, v/v) ml/min Temperature: C UV, nm. Naphthalene. Ethylbenzene R S =.7 0 bar R S =.7 90 bar R S =.7 88 bar 0.0 0..0..0. min Electron microscopic image of NUCLESHELL NUCLESHELL silica particles consist of a non-porous solid core of.7 μm diameter and a porous outer shell of 0. μm thickness. Accordingly the total diameter of the particle is.7 μm. Utilizing a proprietary process of synthesis, NUCLESHELL particles exhibit a distinct narrow particle size distribution (d90/ d0 ~.). Columns packed with NUCLESHELL core shell particles feature exceptional separation efficiencies with theoretical plate numbers easily comparable to totally porous sub micron particles. 9 www.mn-net.com

NUCLESHELL core-shell silica for HPLC Theoretical column efficiency (optimal conditions) Silica d p [µm] L [m] HETP [µm] Efficiency [plates/m] L Analysis [mm] N R s time NUCLESHELL.7 0 000 00 000 % 0 %. 000 0 000 % 0 % NUCLEDUR 7. 0 0 000 00 % 0 %.8. 00 000 0 % 0 %. 80 000 0 0 000 00 % 00 % Benefits of core-shell technology Core-shell particles vs. totally porous silica Short diffusion paths Fast mass transfer (term C of Van Deemter equation) High flow velocity without peak broadening for fast LC Narrow particle size distribution (d 90 /d 0 ~.) Stable packing High heat transfer Minimized influence of frictional heat Efficiency of NUCLESHELL ~ 0 000 m (HETP ~ μm) With conventional fully porous particles the mass transfer between stationary and mobile phase usually results in peak broadening at higher flow rates (C-term in van Deemter equation). The short diffusion paths in the core-shell particles reduce the dwell time of the analyte molecules in the stationary phase, so that even at high flow velocities of the mobile phase, optimal separation results can be obtained. The van Deemter plots demonstrate how efficiency is affected by flow rate. In comparison with fully porous silicas, core-shell particles from various manufacturers maintain the efficiency optimum (max. plates/m) over a long range of increasing linear mobile phase velocity. H = A + B ū -- + C u Van Deemter curves MN Appl. No. 00 Column: 0 x. mm CH CN H (70:0, v/v) Temperature: C Sample: Acenaphthene 0 A term = eddy-diffusion, B term = longitudinal diffusion coeffi cient, C term = mass transfer coeffi cient Plate height h [μm] 8 0 8 NUCLEDUR C 8 Gravity, μm Poroshell 0 EC-C8 NUCLEDUR C 8 Gravity,.8 μm Ascentis Express C8 NUCLESHELL RP 8 Kinetex C8 0 0 8 0 Linear velocity u [mm/s] www.mn-net.com 9

NUCLESHELL core-shell silica for HPLC In direct comparison with conventional sub micron phases, NUCLESHELL columns only generate about 0 % of the back pressure and can be operated with the majority of conventional HPLC systems. In order to develop the maximum performance of NUCLESHELL columns, we recommend reducing extra column voids by using suitable capillaries (< 0. mm inner diameter) and specially adapted detector cells. Moreover detector settings should be optimized by increasing the measuring rate or by decrease of the time constant. P = pressure drop, Φ = fl ow resistance (nondimensional), LC = column length, η = viscosity, u = linear velocity, d P = particle diameter Pressure drop MN Appl. No. 0 Column: 0 x. mm CH CN H (70:0, v/v) Temperature: C Pressure [bar] 00 00 00 00 00 NUCLEDUR C 8 Gravity,.8 μm Poroshell 0 EC-C8 Kinetex C8 Ascentis Express C8 NUCLESHELL RP 8 NUCLEDUR C 8 Gravity, μm 00 0 0 8 0 Linear velocity [mm/s] Core-shell particle technology from MACHEREY-NAGEL is an alternate route to gain highest column efficiency and resolution in HPLC at short run time, but with moderate back pressure. Features of NUCLESHELL particles A criterion for the long-term stability of the column at ph extremes is the percentage decrease of initial retention and initial plates, respectively. The following figure shows a column stability test of NUCLESHELL RP 8 at mobile phase levels ph and ph 0 compared with three competing phases. Stability under acidic and basic conditions MN Appl. Nos. 0 / 0 Columns: 0 x. mm NUCLESHELL RP 8,.7 μm 0 x. mm Kinetex. μm C8 Columns: 0 x. mm NUCLESHELL RP 8,.7 μm 0 x. mm Ascentis Express C8,.7 μm 0 x. mm Poroshell 0 EC-C8 0 x. mm Kinetex, μm C8 acetonitrile % TFA in water, ph (0:0, v/v) 0 mmol/l Na borate 0 mmol/l NaH methanol, ph 0 (:9:0, v/v/v). ml/min. ml/min Temperature: 80 C Temperature: 0 C UV, nm UV, 0 nm Analyt: anthracene Analyt: toluidine % inital retention (anthracene) ph 00 80 0 NUCLESHELL 0 Kinetex 0 0 0 000 0000 000 0000 000 0000 Column volumes Plates (toluidine) 000 ph 0 0000 8000 000 NUCLESHELL 000 Ascentis Express 000 Poroshell 0 Kinetex 0 0 000 0000 000 0000 Column volumes 9 www.mn-net.com

NUCLESHELL core-shell silica for HPLC Columns can be operated at elevated temperatures without loss in retention, efficiency or peak symmetry. Stability test: Column: Temperature stability MN Appl. No. 00 0 x mm NUCLESHELL RP 8,.7 μm A) 0 mmol/l ammonium formate methanol (9:, v/v) + 0 μl formic acid, ~ ph B) 0 mmol/l ammonium formate methanol (:9, v/v) + 0 μl formic acid, ~ ph 0 00 % B in 7 min 0. ml/min, Temperature: 00 C. Phenol. Naphthalene UV, 0 nm Uniformly shaped NUCLESHELL particles combined with optimized bonding technology safeguard tightly packed columns for 00 % reproducible results. Column: Batch-to-batch reproducibility MN Appl. No. 0 0 x mm NUCLESHELL RP 8,.7 μm methanol mmol/l KH P, ph 7 (70:0, v/v) ml/min Temperature: 0 C. Uracil. Toluene. Ethylbenzene. Acenapthene UV, nm. Amitriptyline. o-terphenyl 7. Triphenylene 7 8 h h 0 h h h Batch Batch 0 8 min Effi ciency test: Acetonitrile water (0:0, v/v) 0. ml/min; Temperature: C UV, nm Analyte: Anthracene Batch 0 8 0 min HETP [µm] Asymmetry Start (t = 0). 0.98 End (t = 0 h)..0 www.mn-net.com 9

NUCLESHELL core-shell silica for HPLC Peak capacity The peak capacity is a measure for the number of sample analytes that can be separated on HPLC columns per time unit. Narrow peaks increase the peak capacity and thus the efficiency of the analytical column. n c (normalized) 0 00 0 00 0 0 00 μm μm NUCLEDUR.8 μm.7 μm NUCLESHELL Comparison of peak capacities t g n c = + ( ) W n c : peak capacity t g : gradient time W: peak width (at baseline) The example shows, that in comparison with totally porous NUCLEDUR silica (.8 μm) NUCLESHELL provides % higher peak capacity. Peak capacity MN Appl. No. 0 Columns: 00 x. mm each NUCLESHELL RP 8,.7 μm NUCLEDUR C 8 Gravity,.8 μm NUCLEDUR C 8 Gravity, μm NUCLEDUR C 8 Gravity, μm A) acetonitrile, B) water, 0 00 % A in min. ml/min Temperature: C UV, 0 nm. Acetophenone. Benzoin. Propiophenone. Butyrophenone. Benzophenone. Valerophenone 0 min Max. pressure [bar] Resolution (.) NUCLESHELL,.7 µm. NUCLEDUR,.8 µm 0. NUCLEDUR, µm.97 NUCLEDUR, µm.0 9 www.mn-net.com

NUCLESHELL core-shell silica for HPLC Loading capacity NUCLESHELL columns allow reliable quantification in a wide analytical detection range. Retention time and peak width at 0 % height remain constant with increasing columns load although core-shell particles are suspected of showing a slightly lower loading capacity compared to fully porous silica materials.. 0.8 0. 0. 0. Normalized column parameters Peak width (at 0 % peak height) Retention time 0 0 8 0 Load on column [μg] Column: Loading capacity 0 x mm NUCLESHELL RP 8,.7 μm acetonitrile mmol/l KH P, ph (70:0, v/v) Temperature: 0 C. Valerophenone Area 800 00 00 00 0. ml/min UV, 8 nm R = 0.999 0 0 8 0 Load on column [μg] 9 μg μg 0.9 μg 0. μg 0 min Method transfer of μm particle columns NUCLESHELL is also available in μm particle size to offer all benefits of core-shell technology to all applications which are bound to particle size. Comparison of µm core-shell and totally porous phase Columns: each 00 x. mm A) NUCLESHELL RP 8plus, μm B) NUCLEDUR Gravity C 8, μm methanol water + 0. % formic acid (:, v/v). ml/min Pressure: 8 bar, 9 bar Temperature: C UV, nm Injection:.0 μl Separation of cephalosporin antibiotics MN Appl. No. 0 0.0..0 7. 0.0. min Ret. time [min] Asymmetry (EP) Plates (EP) A B A B A B Cefotaxime.0.9.9. 800 8 Cefoxitin..7..0 99 7 Cefamandole.97.7.. 9 7 Cefalotine..7.. 98 7 www.mn-net.com 97

NUCLESHELL phase overview verview of NUCLESHELL HPLC phases Phase Specification Page Characteristic* Stability Structure RP 8 octadecyl, multi-endcapping 7.8 % C (.7 µm particles). % C ( µm particles) USP L 00 A B C ph, suitable for LC/MS NUCLESHELL (Si- ) n RP 8plus octadecyl (monomeric), multi-endcapping.7 % C (.7 µm particles). % C ( µm particles) USP L 0 A B C - ph 9, suitable for LC/MS NUCLESHELL (Si- ) n Si Si Si Si(CH ) Phenyl-Hexyl phenylhexyl, multi-endcapping. % C (.7 µm particles) USP L 0 A B C ph 0, suitable for LC/MS NUCLESHELL (Si- ) n Si H Si Si(CH ) PFP pentafluorophenyl, multi-endcapping ~ % C (.7 µm particles) USP L 0 A B C ph 9, suitable for LC/MS NUCLESHELL (Si- ) n Si H F F F F F Si Si(CH) HILIC zwitterionic ammonium sulfonic acid. % C (.7 µm particles) 08 A B C - ph 8., suitable for LC/MS NUCLESHELL (Si- ) n CH + N S Si H CH CH + N S Si H CH * A = hydrophobic selectivity, B = polar / ionic selectivity, C = steric selectivity 98 www.mn-net.com

NUCLESHELL phase overview Application Similar phases** Interactions retention mechanism overall sophisticated analytical separations, e.g., analgesics, anti-inflammatory drugs, antidepressants; herbicides; phytopharmaceuticals; immunosuppressants Kinetex C8; Cortecs C8; Raptor C8; Accucore C8; Ascentis Express C8 hydrophobic (van der Waals interactions) Si(CH ) H C CH N overall sophisticated analytical separations, especially for polar compounds, e.g., pharmaceuticals like antibiotics, water-soluble vitamins, organic acids Kinetex XB-C8; Bonshell ASB-C8; Raptor ARC-C8; hydrophobic (van der Waals interactions) Si Si Si Si(CH ) CH N H C aromatic and unsaturated compounds, polar compounds like pharmaceuticals, antibiotics Ascentis Express Phenyl-Hexyl; Kinetex Phenyl-Hexyl; Accucore Phenyl-Hexyl; Ultracore Phenyl-Hexyl; Poroshell Phenyl-Hexyl π-π and hydrophobic N aromatic and unsaturated compounds, phenols, halogenated hydrocarbons, isomers, polar compounds like pharmaceuticals, antibiotics Kinetex PFP; Ascentis Express F; Accucore PFP polar (H bond), dipole-dipole, π-π and hydrophobic F F F H F F H C hydrophilic compounds such as organic polar acids and bases, polar natural compounds ionic / hydrophilic and electrostatic N + S CH CH H N NH H C N + NH S CH ** phases which provide a similar selectivity based on chemical and physical properties www.mn-net.com 99

NUCLESHELL columns NUCLESHELL RP 8 nonpolar high density phase USP L Key feature Core-shell technology for fast and efficient HPLC Suitable for LC/MS and HPLC at ph extremes (ph ) Superior base deactivation, ideal for method development Technical data ctadecyl modification, multi-endcapped; pore size 90 Å, particle size.7 and µm, carbon content 7.8 % for.7 µm,. % for µm; ph stability ; suitable for LC/MS Recommended application verall sophisticated analytical separations, e.g., analgesics, anti-inflammatory drugs, antidepressants; herbicides; phytopharmaceuticals; immunosuppressants NUCLESHELL RP 8 is based on core-shell silica. A unique derivatization process generates a homogeneous surface with a high density of bonded silanes. The following thorough endcapping suppresses any unwanted polar interactions between the silica surface and the sample, which makes NUCLESHELL RP 8 particularly suitable for the separation of basic and other ionizable analytes. The extremely reduced silanol activity of the phase can be demonstrated by applying basic analytes, such as tricyclic antidepressants. The chromatogram below shows a sharp elution profile (superior resolution!) of these highly polar compounds with an excellent asymmetry value for amitriptyline of.. Columns: 0 x. mm each NUCLESHELL RP 8,.7 μm Ascentis Express C8 Kinetex. μm C8 Poroshell 0 EC-C8 methanol acetonitrile mmol/l KH P, ph 7 (.:.:, v/v/v) Pressure: ml/min Temperature: 0 C bar, 9 bar, 8 bar, bar UV, 0 nm Tricyclic antidepressants comparison of selectivity and resolution MN Appl. No. 90. Protriptyline. Desipramine. Maprotiline. Nortriptyline. Doxepin. Imipramine 7. Amitriptyline 8. Clomipramine 9. Trimipramine Asymmetry (amitriptyline) NUCLESHELL.. Ascentis Express.07.9 Kinetex. n.a. Poroshell.0.9 Resolution (8, 9) NUCLESHELL Ascentis Kinetex Poroshell 0 min 7 8 9 0 8 0 8 min 00 www.mn-net.com

NUCLESHELL columns NUCLESHELL RP 8 combines innovative silica technology and excellent surface deactivation, that outperforms conventional C 8 silicas in terms of efficiency, resolution and speed. Due to the applied core-shell particle design the back pressure at elevated flow rates remains at a moderate level and in many cases permits the use of existing HPLC equipment. NUCLESHELL RP 8 with extended ph stability, low bleed characteristics in LC/MS applications, and overall robustness is an ideal tool for method development and routine analyses in modern HPLC. The separation of β-lactam antibiotics illustrates how time of analysis can be shortened to a fractional part by using core-shell particles without loss of resolution at moderate back pressure. Columns: β-lactam antibiotics in less than min MN Appl. No. 90 0 x mm NUCLESHELL RP 8,.7 μm 0 x mm NUCLEDUR C 8 Gravity, μm A) acetonitrile B) 0 mmol/l KH P, ph. 0 % A (0, min) 0 % A in. min (0. min 0 % A) 0 % A ( min) 0 % A in 9 min ( min 0 % A) Pressure: Temperature: C ml/min, ml/min 70 bar, 0 bar UV, 0 nm 0 8 0 min 0 7 8 9 0. Amoxicillin. Ampicillin. Cephalexin. Cefotaxime. Cefoxitin. Cefamandole 7. Cephalothin 8. Piperacillin 9. Penicillin V 0. xacillin. Cloxacillin. Nafcillin. Dicloxacillin. min 70 bar 7 8 9 0.0 0. 0.8...0 min rdering information Eluent in column acetonitrile water ID NUCLESHELL RP 8,.7 µm particle size.7 µm Analytical EC columns Length 0 mm 00 mm 0 mm 0 mm EC guard columns* mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0. mm 7. 7. 7. 78.0 NUCLESHELL RP 8, µm particle size µm Analytical EC columns mm 7.0 7.0 7.0 77.0 78.0 mm 7.0 7.0 7.0 77.0 78.0 mm 7.0 7.0 7.0 77.0 78.0. mm 7. 7. 7. 77. 78.0 EC columns in packs of, guard columns in packs of. Guard column system Guard columns for EC columns with ID mm mm mm. mm Guard column holder * Column Protection System (pack of) EC / () / () / () / () 789 For details of the EC column system please see page 0. www.mn-net.com 0

NUCLESHELL columns NUCLESHELL RP 8plus C 8 phase with polar selectivity USP L Key feature Based on core-shell particle technology for fast and efficient HPLC Hydrophobic C 8 phase with distinct polar selectivity, ideal for method development Excellent performance under highly aqueous conditions Technical data Monomeric octadecyl modification, multi-endcapped; pore size 90 Å, available particle sizes.7 μm and μm, carbon content.7 % for.7 μm,. % for μm; ph stability 9; suitable for LC/MS Recommended application verall sophisticated analytical separations, especially for polar compounds, e.g., pharmaceuticals like antibiotics, water-soluble vitamins, organic acids NUCLESHELL RP 8plus is a C 8 modified core-shell silica. Due to a monomeric bonding chemistry this HPLC phase offers hydrophobic characteristics with distinct polar selectivity. A special derivatization process generates a medium density of bonded silanes with reduced steric selectivity compared to NUCLESHELL RP 8. Column: Bleeding characterisitics MN Appl. No. 0 0 x mm NUCLESHELL RP 8plus,.7 μm A) 0. % formic acid in water B) 0. % formic acid in acetonitrile 9 % A % A in. min (0. min) 9 % A in 0. min (. min) 0. ml/min Temperature: C Total ion chromatogram (TIC), 0 000 m/z MS NUCLESHELL RP 8plus NUCLESHELL RP 8 Poroshell C8 Kinetex XB-C8 Blank 0 7 8 9 0 Retention time [min] NUCLESHELL RP 8plus combines superbly hydrophobic and polar selectivity so it is a useful tool for method development in RP chromatography. Good ph stability and low bleeding characteristics make it ideal especially for LC/MS applications. Column: Eluent ph : Eluent ph 9: ph stability of NUCLESHELL RP 8plus MN Appl. No. 0 00 x mm NUCLESHELL RP 8plus,.7 μm % TFA in water - acetonitrile (0:0, v/v) 0 mmol/l triethylammonium acetate adjusted to ph 9 for ph : 0.8 ml/min, for ph 9: 0. ml/min Temperature: for ph : 0 C, for ph 9: 0 C Injection: Retention time (relative to initial value) Plate number (relative to initial value) 0 % 0 % 00 % 9 % 90 % 0 % 0 % 00 % 9 % UV, nm μl Phenol ph H 0 8 0 8 0 h Anthracene ph 9 90 % 0 8 0 8 0 8 h Also a comparison of retention of the glycopeptide antibiotic vancomycin on several octadecyl modified core-shell phases underlines the polar selectivity of NUCLESHELL RP 8plus. 0 www.mn-net.com

NUCLESHELL columns Columns: Temperature: C Injection: Polar selectivity shown for vancomycin MN Appl. No. 0 0 x mm each NUCLESHELL RP 8plus,.7 μm NUCLESHELL RP 8,.7 μm Kinetex. μm C8 water methanol acetonitrile glacial acetic acid (00:8::0., v/v/v/v) adjusted to ph. with sodium hydroxide solution 0.9 ml/min UV, 0 nm 0 μl H H NH NH H H N N N N H H H HN H N H H H In addition NUCESHELL RP 8plus provides a good stability under highly aqueous conditions. Even by long term usage or storage of the phase phase collapse and loss of retention are hardly observed. The original performance can be regained after a short regeneration procedure. Column: Phase collapse and regeneration MN Appl. No. 70 00 x mm NUCLESHELL RP 8plus,.7 μm 0 mmol/l KH P, ph. 0. ml/min Temperature: 0 C Injection: UV, nm 0. μl 8 h storage in water New column 0 min regeneration H Cl H H H N H Cl Fumaric acid H H 0 min 0 min rdering information Eluent in column acetonitrile water ID NUCLESHELL RP 8plus,.7 µm particle size.7 µm Analytical EC columns Length 0 mm 00 mm 0 mm 0 mm EC guard columns* mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0. mm 7. 7. 7. 78.0 NUCLESHELL RP 8plus, µm particle size µm Analytical EC columns mm 7.0 7.0 7.0 77.0 78.0 mm 7.0 7.0 7.0 77.0 78.0 mm 7.0 7.0 7.0 77.0 78.0. mm 7. 7. 7. 77. 78.0 EC columns in packs of, guard columns in packs of. Guard column system Guard columns for EC columns with ID mm mm mm. mm Guard column holder * Column Protection System (pack of) EC / () / () / () / () 789 For details of the EC column system please see page 0. www.mn-net.com 0

NUCLESHELL columns NUCLESHELL Phenyl-Hexyl nonpolar high density phase USP L Key feature Based on core-shell particle technology for fast and efficient HPLC Hydrophobic phase with alternative selectivity compared to classical C 8 modifications Separation principle based on retention mechanisms: π-π interactions and hydrophobic interactions Technical data Phenyl-Hexyl modification, multi-endcapped; pore size 90 Å, particle size.7 μm; carbon content. %; ph stability 0; suitable for LC/MS Recommended application Aromatic and unsaturated compounds, polar compounds like pharmaceuticals, antibiotics Phenyl-Hexyl modified phases offer an excellent separation efficiency especially for aromatic and unsaturated compounds with electron-withdrawing groups. The combination of hydrophobic and π-π interactions results in an alternative and interesting selectivity profile compared to C 8 or C 8 modifications. NUCLESHELL Phenyl-Hexyl is based on a unique surface bonding chemistry - therefore it is suitable for LC/MS due to low bleeding characteristics and offers high temperature stability and ph stability from to 0. Column: Stability of NUCLESHELL Phenyl-Hexyl at ph 0 MN Appl. No. 0 0 x mm NUCLESHELL Phenyl-Hexyl,.7 μm acetonitrile 0 mmol/l TEA ph 0 (0:0, v/v); ph of the mixture 0. ml/min Temperature: 0 C Injection:. Phenol. Naphthalene. Anthracene UV, nm μl Columns: Bleeding characteristics of NUCLESHELL Phenyl-Hexyl cps.0e8.0e7 0 MN Appl. No. 00 0 x mm each NUCLESHELL Phenyl-Hexyl,.7 μm Kinetex Phenyl-Hexyl A) acetonitrile, B) water 9 % A in min 0. ml/min Temperature: C MS 8 0 min The pyridine-phenol test shows that NUCLESHELL Phenyl-Hexyl provides a symmetrical peak for pyridine and higher resolution in comparison to other core-shell based Phenyl-Hexyl phases, which underlines the excellent base deactivation. Pyridine-phenol test of NUCLESHELL Phenyl-Hexyl % 00 80 0 0 h 0 h 0 min Relative plate numbers MN Appl. No. 0 Columns: 0 x mm each NUCLESHELL Phenyl-Hexyl,.7 μm Kinetex Phenyl-Hexyl Ascentis Express Phenyl-Hexyl acetonitrile water (70:0, v/v) 0. ml/min Temperature: 0 C UV, nm Injection: 0. μl 0 0 0 0 0 0 h NUCLESHELL Phenyl-Hexyl is a robust phase with an alternative RP selectivity for aromatic and unsaturated analytes compared to classical C 8 / C 8 phases it is an additional and useful tool for all chromatography users.. Pyridine. Phenol 0.0 0..0 min 0 www.mn-net.com

NUCLESHELL columns Columns: Temperature: 0 C Injection: Comparing the separation of sulfonamides on NUCLEDUR Phenyl-Hexyl with different particle sizes 0 x mm each NUCLESHELL Phenyl-Hexyl,.7 μm NUCLEDUR Phenyl-Hexyl,.8 μm NUCLEDUR Phenyl-Hexyl, μm NUCLEDUR Phenyl-Hexyl, μm A) methanol B) 0. % formic acid in water 0 80 % A in 0 min 0. ml/min UV, nm 0. μl MN Appl. No. 80 R s. 7 R s.0. Sulfadiazine. Sulfachlorpyridazine. Sulfapyridine. Sulfamerazine. Sulfadimidine. Sulfathiazole 7. Sulfadimethoxine n NUCLESHELL Phenyl-Hexyl the resolution of the last two peaks is higher than on the fully porous.8 μm NUCLEDUR Phenyl-Hexyl. R s.8 R s. 0 8 min The separation of sulfonamides proves the scalability from fully porous NUCLEDUR to NUCLESHELL Phenyl-Hexyl. Hereby the core-shell silica exhibits identical selectivity, narrower peaks and slightly shorter retention under the same conditions. Thus, method transferability between NUCLEDUR and NUCLESHELL is guaranteed, either for speeding up your methods or scaling up for preparative requirements. rdering information Eluent in column acetonitrile water ID Length 0 mm 00 mm 0 mm EC guard columns* NUCLESHELL Phenyl-Hexyl,.7 μm particle size.7 µm Analytical EC columns mm 77.0 77.0 77.0 778.0 mm 77.0 77.0 77.0 778.0 mm 77.0 77.0 77.0 778.0. mm 77. 77. 77. 778.0 EC columns in packs of, guard columns in packs of. Guard column system Guard columns for EC columns with ID mm mm mm. mm Guard column holder * Column Protection System (pack of) EC / () / () / () / () 789 For details of the EC column system please see page 0. www.mn-net.com 0

NUCLESHELL columns NUCLESHELL PFP hydrophobic pentafluorophenyl phase USP L Key feature Core-shell technology for fast and efficient HPLC Hydrophobic phase with alternative selectivity in comparison to classical C 8 modifications Separation principle based on retention mechanisms (polar interactions (H bonds), dipole-dipole, π-π, hydrophobic interactions) Technical data Phase with pentafluorophenylpropyl modification, multi-endcapping; pore size 90 Å, particle size.7 μm; carbon content ~ %; ph stability 9; suitable for LC/MS Recommended application Aromatic and unsaturated compounds, phenols, halogen compounds, isomers, polar compounds like pharmaceuticals, antibiotics; strong retention of basic compounds rthogonality in selectivity Fluorinated stationary phases in HPLC have gained increasing interest over the last years. Most common representative of fluorinated silica phases is the pentafluorophenyl modification (PFP or F ). Especially the orthogonal selectivity compared to traditional alkyl phases widens the scope in analytical HPLC. Thus NUCLESHELL PFP offers an excellent selectivity especially for highly polar analytes, aromatic and unsaturated compounds, phenols or halogenated hydrocarbons. While a typical C 8 phase just provides hydrophobic interactions between stationary phase and analyte NUCLESHELL PFP offers four different retention mechanisms: polar interactions (H bonds), dipole-dipole interactions, π-π interactions and hydrophobic interactions. Especially the pronounced ion exchange capacity and distinct steric selectivity are typical for the character of fluorinated phases. Columns: Temperature: 0 C Sample: 0 00 80 0 Stability of NUCLESHELL PFP at ph MN Appl. No. 0 00 x. mm NUCLESHELL PFP,.7 μm 00 x. mm Kinetex PFP,. μm F acetonitrile 0. % TFA, ph (0:0, v/v). ml/min UV, nm ethylbenzene NUCLESHELL PFP % initial retention 0 % initial plates 0 0 0 000 000 000 Column volumes β-blockers orthogonal selectivity of NUCLESHELL PFP MN Appl. No. 0 Columns: 00 x. mm NUCLESHELL RP 8,.7 μm NUCLESHELL PFP,.7 μm A) acetonitrile + 0. % formic acid B) 0. % formic acid 0 % A in. min, -0 % A in min.7 ml/min Temperature: C UV, 80 nm. Atenolol. Labetalol. Pindolol. Alprenolol. Metroprolol. Propranolol 0 min 0 www.mn-net.com

NUCLESHELL columns Columns: 00 x. mm NUCLESHELL PFP,.7 μm 0 x mm NUCLEDUR PFP, μm 00 x. mm Kinetex. μm F Methanol water (:, v/v). ml/min, ml/min, ml/min,. ml/min Temperature: C UV, nm Methylacetophenones MN Appl. No. 90. o-methylacetophenone. p-methylacetophenone. m-methylacetophenone Kinetex F,. ml/min NUCLESHELL PFP ml/min NUCLESHELL PFP,. ml/min 0 0 min NUCLEDUR PFP, ml/min 0 0 0 0 0 min NUCLESHELL PFP combines the benefits of core-shell technology, high stability, and orthogonal selectivity. Thus it is a useful complementary tool for highly efficient separations especially of isomers, halogenated, aromatic and / or polar compounds. rdering information Eluent in column acetonitrile water ID Length 0 mm 00 mm 0 mm EC guard columns* NUCLESHELL PFP,.7 µm particle size.7 µm Analytical EC columns mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0. mm 7. 7. 7. 78.0 EC columns in packs of, guard columns in packs of. Guard column system Guard columns for EC columns with ID mm mm mm. mm Guard column holder * Column Protection System (pack of) EC / () / () / () / () 789 For details of the EC column system please see page 0. www.mn-net.com 07

NUCLESHELL columns NUCLESHELL HILIC zwitterionic phase Key feature Core-shell technology for fast and efficient HPLC Ideal for reproducible and stable chromatography of highly polar analytes Very short column equilibration times Technical data Ammonium - sulfonic acid modified silica; pore size 90 Å, particle size.7 μm; carbon content. %; ph stability 8.; suitable for LC/MS Recommended application Hydrophilic compounds such as polar organic acids and bases, polar natural compounds, nucleosides, oligonucleotides, amino acids, peptides, water-soluble vitamins Hydrophilic interaction chromatography Good separation of polar compounds like the physiologically important substances creatine and creatinine can be achieved on NUCLESHELL HILIC as well as on NUCLEDUR HILIC,.8 μm at similar retention, but much lower back pressure. NPC adsorbent HILIC eluent RPC analyte IC Columns: Separation of creatine and creatinine MN Appl. No. 990 0 x mm NUCLESHELL HILIC,.7 μm 0 x mm NUCLEDUR HILIC,.8 μm acetonitrile 0 mmol/l ammonium acetate, ph.0 (90:0, v/v) Pressure:.7 ml/min 9 bar 80 bar Temperature: C UV, 0 nm Hydrophilic interaction chromatography (HILIC) is a separation technique using polar stationary phases and organic-aqueous mobile phases. A minimum water content of at least % is indispensable to provide a permanent water layer between the adsorbent surface and the organic fraction of the mobile phase. The sample molecules become separated in a partition chromatography, in which polar analytes are more strongly retained than neutral, less hydrophilic compounds. Consequently, increasing the aqueous part in the mobile phase will diminish retention of the polar sample constituents. In this way HILIC behaves inverse to classical RP chromatography. The particular retention profile of HILIC enables the chromatography of very polar and often small molecules, which won t show any retention on C 8 or C 8 reversed phases. Ultra-fast separations at moderate back pressure NUCLESHELL HILIC is a core-shell technology based stationary phase with a covalently bonded -N,N-dimethylaminopropane sulfonic acid ligand (pat. p nd.). The betaine character of the strong ion-exchanger results in full charge balancing and facilitates fast equilibration times.. Creatinine. Creatine 0.0 0. 0. 0. 0.8 min The following chromatograms show the method transfer from a fully porous μm HILIC phase to.7 μm core-shell silica with equal selectivity features. Run time has been cut down to min. Column back pressure remains modest < 00 bar, while solvent demand is reduced to less than %. Si CH + N S CH 08 www.mn-net.com

NUCLESHELL columns Separation of catecholamines MN Appl. No. 0 Columns: 00 x mm NUCLESHELL HILIC,.7 μm 00 x mm NUCLESHELL HILIC,.7 μm min ml. min. ml min ml (solvent consumption per run) Pressure: 0 x mm NUCLEDUR HILIC, μm acetonitrile 00 mmol/l ammonium formate, ph. (80:0, v/v) ml/min, ml/min, ml/min 9 bar, 9 bar, bar Temperature: C UV, 80 nm. DPAC. Serotonin. Norepinephrine. DPA. Dopamine. Epinephrine 7. DPS 7 7 Core-shell { 7 Fully porous 0 0 min Core-shell silica: separation in min pressure < 00 bar NUCLESHELL HILIC provides stable and reproducible chromatography, comprising all the benefits of a state-of-the-art core-shell silica. rdering information Eluent in column acetonitrile water ID Length 0 mm 00 mm 0 mm EC guard columns* NUCLESHELL HILIC,.7 µm particle size.7 µm Analytical EC columns mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0 mm 7.0 7.0 7.0 78.0. mm 7. 7. 7. 78.0 EC columns in packs of, guard columns in packs of. Guard column system Guard columns for EC columns with ID mm mm mm. mm Guard column holder * Column Protection System (pack of) EC / () / () / () / () 789 For details of the EC column system please see page 0. www.mn-net.com 09