InertSustain AQ-C18. HPLC, LC/MS Columns. Maximizing retention for highly polar compounds in reversed phase methods with highly aqueous mobile phases

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1 HPLC, LC/MS Columns InertSustain AQ-C8 Maximizing retention for highly polar compounds in reversed phase methods with highly aqueous mobile phases Physical Properties Silica : ES (Evolved Surface) Silica Gel Particle Size :.9 μm, μm, μm Surface Area : 0 m /g Pore Size : 00 Å (0 nm) Pore Volume : 0.8 ml/g Bonded Phase : Octadecyl Groups End-capping : Complete Carbon Loading :.0 % USP Code : L ph Range : ~0 Authorised Distributor: Hichrom Limited sales@hichrom.co.uk

2 InertSustain AQ-C8 Benefits. Exceptional retention for highly polar compounds (hydrophilic). Highly inert packing material results in less tailing of peaks for virtually any type of analytes. Extreme resistance to low and high ph mobile phases. Endlessly reproducible from column-to-column and batch-to-batch InertSustain AQ-C8 columns are designed to achieve strong retention for highly polar compounds, which is the most challenging goals in developing reversed phase methods. The optimization of bonding of the C8 groups at equal distance to the silica gel enable InertSustain AQ-C8 to offer significant retention for highly polar compounds even under water rich mobile phases. As illustrated in the following plot, InertSustain AQ-C8 provided exceptional retention for highly polar compounds even under water rich mobile phases without dewetting or phase collapse. Retention Properties of InertSustain AQ-C8 Retention Factor of acidic, basic and neutral compounds using 00% water mobile phase InertSustain AQ-C8 Aquasil C8 Atlantis T Inertsil ODS- Inertsil ODS-SP Inertsil ODS- InertSustain C8 CAPCELL PAK C8-AQ Synergi Fusion RP XSelect CSH C8 Retention Factor of amylbenzene using methanol/water=80/0 Remarks:. The horizontal and vertical retention factor values were calculated with reference to the retention factor values obtained from InertSustain AQ-C8.. The horizontal values represents the retention factor of amylbenzene using methanol/water=80/0.. The vertical values represents the retention factor of acidic, basic and neutral compounds using 00% water mobile phase. (For further details, please see the following chart) InertSustain AQ-C8 provided strong retention for all basic, neutral and acidic compounds under 00% water mobile phase Remarks: -methyluridine Neutral Pyridoxamine Basic Dopamine Generally, the retention for acidic compounds become stronger than basic compounds when the surface of the silica gel is basic. The retention for basic compounds become stronger than acidic compounds when the surface of the silica gel is acidic. InertSustain AQ-C8 Uracil Succinic Acid Acidic Acetic Acid CAPCELL PAK C8-AQ Atlantis T Synergi Fusion RP

3 Highly Polar Compound Retention with 00% Aqueous Mobile Phases It is indeed difficult to retain highly polar samples by reversed phase mode as the polar samples tend to elute near the t 0 (void volume) with water rich mobile phases. On the other hand, polar embedded C8 columns or C8 columns with hydrophilic endcapping are available in the market for retaining highly polar compounds. However, they often show tailing or poor peak shapes due to the secondary interaction from their embedded functional groups. InertSustain AQ-C8 not only provide strong retention for highly polar compounds, but also delivery symmetric peak shapes for virtually any type of analytes. InertSustain AQ-C8 Atlants T Synergi Fusion RP t0 t0 t0 + : Not eluted Column : μm, 0. mm I.D. : 0. % HCO in H O Flow Rate :.0 ml/min Col. Temp. : 0 Detection : UV 0 nm. Pyridoxamine (Vitamin B). Thiamin (Vitamin B). Nicotinic acid (Vitamin B). Pyridoxal (Vitamin B). Nicotinamide (Vitamin B). Pyridoxine (Vitamin B) InertSustain C8 (Conventional C8 column) t Time(min) Retentivity Under 0% Organic Solvent As shown below, InertSustain AQ-C8 delivered stronger retention of catechin sample even under 0% organic solvent mobile phase with exceptional peak shapes, while competitive column brand failed. Furthermore, InertSustain AQ-C8 can prevent the co-elution between the targeted polar analytes and solvent peaks or sample matrices due to its enhanced retentivity. InertSustain AQ-C8 Atlants T Complete separation between solvent peak Elution with solvent peak Poor peak shape Column : μm,0. mm I.D. : A) 0. %HCO in H O B) CH CN A/B= 80/0, v/v Flow Rate :.0 ml/min Col. Temp. :0 Detection :UV 80 nm. Gallocatechin (-) (GC). Catechin (+) (C). Epicatechin (-) (EC). Epigallocatechin gallate (-) (EGCg). Epicatechin gallate (-) (ECg). Catechin gallate (-) (Cg) 0 8 0

4 InertSustain AQ-C8 Applications High-Throughput Analysis of Drugs InertSustain AQ C8 (.9 μm) Acquity BEH C8 (.7 μm).0 (x,000,000)..0. Increase in Sensitivity.0 (x,000,000) min min Column Size Flow Rate Detection Injection Vol. : 0. mm I.D. : A) 0.0 % HCO in H O : B) CH CN A/B= 70/0 - (0. min) - 0/90 - (0. min hold) : 0. ml/min : LC/MS/MS(ESI,SRM) : μl. Propranolol(+). Carbamazepine(+). Salicylic acid(-). Ketoprofen(-) (each 00 μg/l in 0 % ACN) (+): Positive mode (-): Negative mode Analysis of Nucleotide in Fish Meat mau 0 8 mau Column : InertSustain AQ-C8 ( μm, 0. mm I.D.) : 0 mm K HPO in H O (ph 7.0, H PO )* Flow Rate :.0 ml/min Col.Temp. : 0 Detection : UV 0 nm Injection Vol. : μl.imp.atp.adp.amp.hyp.ino (each mg/l) * Equilibrate the column with CH CN/H O=/,v/v after the analysis. When storing the column for a long period of time, store it with 00 % CH CN 00 %.

5 Applications Analysis of Water-Soluble Vitamins.9 μm, 0. mm I.D. 0. ml/min μm, 0. mm I.D.0 ml/min Column Col. Temp. Detection :InertSustain AQ-C8 :A)0.% H PO B) CH CN* :0 :UV 0 nm 8 *Gradient Time Program <.9 μm> Time (min) A% B% < μm> Time (min) A% B% Pyridoxamine. Thiamin. Nicotinic Acid. Ascorbic acid. Nicotinamide. Pyridoxal 7. Pyridoxine 8. Pantonenic acid 9. Folic Acid 0. Cyanocobalamin. Riboflavin. Biotin Analysis of Organic Acids 7 Column : μm, 0. mm I.D. : 0 mm NaH PO in H O (ph., H PO ) Flow Rate :.0 ml / min Col. Temp. :0 Detection :UV 0 nm Inj. Vol. :0 μl Oxalic acid 0 mg/l. Tartaric acid 00 mg/l. Formic acid 000 mg/l. Malic acid 000 mg/l. Lactic acid 000 mg/l. Acetic acid 000 mg/l 7. Maleic acid 0 mg/l 8. Citric acid 000 mg/l 9. Pyroglutamic acid 00 mg/l 0. Succinic acid 000 mg/l. Fumaric acid 0 mg/l 0 8

6 InertSustain AQ-C8 Applications Analysis of Catecolamines Column : μm, 0. mm I.D. : 0.% H PO in H O Flow Rate :.0 ml / min Col. Temp. :0 Detection :UV 0 nm Inj. Vol. : μl. Norepinephrine. L-Adrenaline. Dopamine. L-DOPA. L-(-)-Tyrosine HO NH HO N H NH HO O O HO NH HO NH Analysis of Fish Freshness, K-Value Column : μm, 0. mm I.D. : 0 mm NaH PO in H O Flow Rate :.0 ml / min Col. Temp. :0 Detection :UV nm Inj. Vol. : μl Sample. ATP. ADP. IMP. Hypoxanthine. AMP. Inosine :00 ppm 0 0 0

7 Applications Analysis of β blocker Column : μm, 0. mm I.D. : A: CH CN B: 0.% H PO in H O A/B=/7, v/v Flow Rate :.0 ml / min Col. Temp. :0 Detection :UV 0 nm Inj. Vol. : μl. Atenolol. Acebutolol. Oxprenolol. Labetalol. Propranolol. Alprenolol (00 μg/ml each) Analysis of Nucleoside and Nucleic Acid Base Column : μm, 0. mm I.D. : 0. M KH PO in H O + 0. M NaClO in H O (ph.0, H PO ) Flow Rate :.0 ml / min Col. Temp. :0 Detection :UV 0 nm Inj. Vol. : μl mau Cytosine. Uracil. Guanine. Adenine. Cytidine. Uridine 7. Thymine 8. Adenosine 9. Guanosine 0. Thymidine (00 mg/l each)

8 InertSustain AQ-C8 Applications Analysis of Nucleotides via LC/MS/MS Samples such as nucleotides have several phosphate groups which is sensitive to stainless steel hardware. As shown below, the combination of highly inert packing material of InertSustain AQ-C8 and usage of a new *Steel- Coated-PEEK hardware (metal-free) deliver excellent peak shapes with higher sensitivity for phosphate compounds WITHOUT the formation of phosphate-iron complexes found with stainless steel column hardwares. *Ordering information on Steel-Coated-PEEK hardware is available at page 0. System Column size : Nexera LCMS-800 plus system : μm HP, 0 x. mm I.D : A) mm Ammonium formate in H O B) CH CN Flow Rate : 0. ml/min Col. Temp. : 0 Detection : LC/MS/MS (ESI), Positive Injection Vol. : μl B% Sample :.ATP 00 μg/l.adp 00 μg/l.amp 00 μg/l Steel-Coated-PEEK hardware Packed with InertSustain AQ-C8 Stainless Steel Hardware Packed with InertSustain AQ-C8 PEEK Packing Material Packing Material Stainless Steel Stainless Steel.0 (x0,000) RT=. min..0 (x0,000). RT=. min Adsorption to the stainless steel hardware.0. Higher Sensitivity without adsorption t0 0. t min min 8

9 Ordering Information Analytical Columns Particle Size:.9 μm 80 MPa (800 Bar) End-fittings are UP Type end-fittings. Length/ I.D. (mm) End-fitting Format. mm UP Type End-fittings HP Series Particle Size: μm 0 MPa (00 Bar) Length/ I.D. (mm) End-fittings are / Waters-compatible. UHPLC compatible end-fittings are also availa ble upon request for UHPLCsystems (Ex: UPLC )to avoid dead volume. Indicate UP Type end-fittings when ordering. (Please note that UP type is not available for a. mm I.D. column). mm / Waters End-fittings UPLC is a registered trademark of Waters Corporation. Particle Size: μm 0 MPa (00 Bar) Particle Size: μm 0 MPa (00 Bar) Length/I.D. (mm) Length/I.D. (mm) Length/I.D. (mm) Length/I.D. (mm) End-fittings are / Waters-compatible. Other column sizes available upon request. 9

10 InertSustain AQ-C8 Ordering Information Guard Column for UHPLC (Max. Operating Pressure 80 MPa, 800 Bar) I.D. of the Analytical Column Applicable (mm).0 Length (mm) I.D. (mm) Replacement Cartridge ( pcs) For connection details, please refer to the diagram below. Cartridge ( pcs) + Holder ( pcs) Set Replacement Cartridge ( pcs) Cartridge ( pcs) + Holder ( pcs) Set Particle Size.9 μm Particle Size.9 μm Particle Size μm Particle Size μm , , Holder for Guard Column for UHPLC Instrument Side / (No. 0- UNF) Holder PEEK Guide Packing Connection to Analytical Column Cartridge Guard Column / O.D. x 0.8 mm I.D. x 0 mm tubing Cartridge Guard Columns I.D. of the Analytical Column Applicable (mm).0 Length (mm) I.D. (mm) End-fittings are / Waters-compatible. Maximum operating pressure is 0 MPa, 00 Bar. Replacement Cartridge E ( pcs) Particle Size Cartridge E ( pcs) + Holder ( pcs) Set Particle Size μm μm μm μm , , , , , Holder for Cartridge Guard Column E Preparative Columns For 0 mm Length For 0 mm Length Particle Size: μm 0 MPa (00 Bar) Length/ I.D. (mm) Preparative Guard Columns Particle Size: μm 0 MPa (00 Bar) Length x I.D. (mm)

11 Ordering Information Capillary Columns EX-Nano Columns Particle Size: μm MPa (0 Bar) EX Columns Particle Size: μm 0 MPa (00 Bar) EX-Nano Columns Particle Size: μm MPa (0 Bar) EX Columns Particle Size: μm 0 MPa (00 Bar) Length/ I.D. (mm) Length/ I.D. (mm) Length/ I.D. (mm) Length/ I.D. (mm) End-fittings are / Valco (0- UNF). / Valco (-0 UNF) end-fittings ca n also be arranged upon request. Indicate / end-fittings when ordering. Connection Kits for Capillary EX-Nano and EX Columns Column Coupler mm I.D. / O.D. Tubing (Both ends with male nuts including PEEK ferrules) Capillary Tubing Connector mm I.D. 0.7 mm O.D. Tubing mm I.D. 0.7 mm O.D. Tubing mm I.D. 0.7 mm O.D. Tubing (Male nut, PEEK ferrule, / O.D. PTFE with sleeve) For Capillary EX-Nano Columns PTFE Tubing 0 mm pcs / O.D. (O.D. 0.7 mm Connection for Capillary Tubing) Column Coupler 0 0. mm I.D. / O.D.Tubing (Both ends with male nuts including PEEK ferrules) Capillary Tubing Connector mm I.D. 0.7 mm O.D. Tubing (Male nut, PEEK ferrule, / O.D. PTFE with sleeve) For Capillary EX Columns PTFE Tubing 0 mm pcs / O.D. (O.D. 0.7 mm Connection for Capillary Tubing)

12 InertSustain AQ-C8 Ordering Information Metal-Free PEEK Columns PEEK Columns Particle Size: μm 0 MPa (00 Bar) Steel-Coated PEEK Columns Particle Size: μm 0 MPa (00 Bar) Length/ I.D. (mm).. 0 Please inquire - Please inquire Please inquire 0 Please inquire Please inquire 7 - Please inquire 00 Please inquire Please inquire 0 Please inquire Please inquire 00 - Please inquire 0 Please inquire Please inquire 00 Please inquire Please inquire Length/ I.D. (mm).. 0 Please inquire Please inquire 00 Please inquire Please inquire 0 Please inquire Please inquire 0 Please inquire Please inquire End-fitting Format. mm UP Type End-fittings End-fittings are UP type end-fittings. For further product details, please contact your local GL Sciences representative. To order: Tel: + (0) sales@hichrom.co.uk

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