Separation of Structurally Similar Steroids on HALO C18 and PFP

Transcription

1 Application Note: 047-STR Separation of Structurally Similar Steroids on HALO C8 and PFP, 4 G009 HALO C8 HALO PFP 4. Prednisone. Cortisone. Prednisolone 4. Hydrocortisone Columns: 4.6 x 00 mm, HALO C8 4.6 x 00 mm, HALO PFP Part Numbers: C8, PFP, Mobile Phase: 50/50: water/methanol Flow Rate:.0 ml/min. Pressure: About 0 Bar Temperature: 5 C Detection: UV 40 nm, VWD Injection Volume: 0.5 µl Sample Solvent: 80% methanol in water Response Time: 0.0 sec. Flow Cell:.5 µl semi-micro ECV: ~4 µl Prednisone Prednisolone Cortisone Hydrocortisone The unique selectivity of HALO PFP is useful in the separation of the closely related steroids prednisolone and hydrocortisone. The electron-deficient ring structure of the perfluorophenyl group aids in separating compounds through pi-pi interactions with the sample.

2 Application Note: 57-AM Isocratic Separation of Amphenicols on HALO Phenyl-Hexyl Phase G009. Thiamphenicol. Chloramphenicol 40 nm 80 nm Column: 4.6 x 50 mm, HALO Phenyl-Hexyl Part Number: Mobile Phase: 55/45-A/B A= 0.05 M Ammonium acetate buffer, ph=5.8 B=Acetonitrile Flow Rate:.0 ml/min. Pressure: 94 Bar Temperature: 5 C Detection: UV 40/80 nm, VWD Injection Volume: 0. μl Sample Solvent: Acetonitrile Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro Extra column volume: ~4 μl Thiamphenicol Chloramphenicol This separation shows a rapid HPLC method for the analysis of amphenicols on HALO Phenyl-Hexyl stationary phase. To improve the sensitivity of detection the first peak was 40 nm and the 80 nm. :

3 Application Note: 5-BZ Separation of Benzodiazepines on HALO Phenyl-Hexyl, C8, and PFP Phases STRUCTUR G00A G00B G00C. Oxazepam. Lorazepam. Nitrazepam 4. Alprazolam 5. Clonazepam 6. Temazepam 7. Flunitrazepam 8. Diazepam Columns: 4.6 x 50 mm, HALO Phenyl-Hexyl, C8, and PFP Part Numbers: , , and , respectively Mobile Phase: A= 5 mm Ammonium acetate in water (ph=5.8 not adjusted), B= Acetonitrile Gradient from 4-6% B in.5 minutes Flow Rate:.5 ml/min. Pressure: 00 Bar Temperature: 5 C Detection: UV 54 nm, VWD Injection Volume:.0 μl Standard diluted with Acetonitrile and buffer Response Time: <0. sec. Flow Cell: 5 μl semi-micro LC System: Agilent 00 Gradient dwell volume= 0.88 ml Oxazepam Lorazepam Alprazolam Clonazepam Flunitrazepam Diazepam These separations of benzodiazepines on three different HALO Fused-Core HPLC stationary phases show the utility of having a variety of phases to optimize selectivity and/or to shorten analysis time. Nitrazepam Temazepam

4 Application Note: 04-ST Separation of Statin Drugs on HALO C8 G006 4 STRUCTUR. Pravastatin. Atorvastatin. Mevastatin 4. Simvastatin Column: 4.6 x 50 mm, HALO C8 Part Number: Mobile Phase: 0/80: A/B A= 0.0 M formic acid in water B= 0.0 M formic acid in methanol Flow Rate:.0 ml/min. Pressure: 40 Bar Temperature: 0 C Detection: UV 40 nm, VWD Injection Volume:.0 μl Sample Solvent: mobile phase Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro ECV: ~4 μl Pravastatin Mevastatin The statin drugs can be rapidly separated using short HALO C8 columns. Atorvastatin Simvastatin

5 Application Note: 65-B Separation of Antiulcer Drugs on HALO Penta-HILIC G0047 (54 nm) 4 STRUCTUR. Cimetidine. Nizatidine. Famotidine 4. Ranitidine Column: 4.6 x 00 mm, HALO Penta-HILIC Part Number: Mobile Phase: 0/90: A/B A= 0.04 M Ammonium formate, ph=.0 (adj.) B= Acetonitrile Flow Rate:.0 ml/min. Pressure: 0 Bar Temperature: 0 C Detection: UV 54 nm, VWD Injection Volume: μl Sample Solvent: mobile phase Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro ECV: ~4 μl The strongly basic antiulcer drugs can be rapidly separated on HALO Penta-HILIC phase using a mobile phase that works well with a mass spectrometer detector. Cimetidine Nizatidine Famotidine Ranitidine

6 Application note: 64-B Separation of 5 Beta Blocker Drugs on HALO Penta-HILIC (54 nm) 4 G0046 STRUCTUR 5. Alprenolol. Propranolol. Pindolol 4. Acebutolol 5. Atenolol Column: 4.6 x 00 mm, HALO Penta-HILIC Part Number: Mobile Phase: 0/90: A/B A= 0.04 M Ammonium formate buffer, ph=.0 B= Acetonitrile Flow Rate:.0 ml/min. Pressure: 5 Bar Temperature: 0 C Detection: UV 54 nm, VWD Injection Volume:.0 μl Sample Solvent: mobile phase Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro ECV: ~4 μl Alprenolol Propranolol Pindolol Acebutolol The HALO Penta-HILIC stationary phase can rapidly separate highly basic compounds with good peak shapes in a mass spectrometry friendly mobile phase. Atenolol

7 Application Note: 60-CB Separation of Carbamate Pesticides on HALO ES-CN Phase G004. Carbetamide. Propham. Chlorpropham Column: 4.6 x 50 mm, HALO ES-CN Part Number: Mobile Phase: 40/60-A/B A= Water B=Acetonitrile Flow Rate:.0 ml/min. Pressure: 65 Bar Temperature: 0 C Detection: UV40 nm, VWD Injection Volume: 0. μl Sample Solvent: Acetonitrile Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro Extra column volume: ~4 μl Carbetamide Propham This separation illustrates a rapid HPLC determination of three carbamate pesticides on HALO ES-CN phase in just over half a minute. The unique Fused-Core technology allows the use of high flow rates at moderate pressures while retaining high efficiency. Chlorpropham :

8 Application Note: 54-P Isocratic Separation of Phenyl Ureas on HALO ES-CN G006. Fenuron. Monuron. Fluomethuron 4. Diuron 5. Linuron 6. Neburon Column: 4.6 x 50 mm, HALO ES-CN Part Number: Mobile Phase: 50/50-A/B A=0.0 M Phosphate buffer, adj. to ph=.5 B=Acetonitrile Flow Rate:.0 ml/min. Pressure: 00 Bar Temperature: 0 C Detection: UV 45 nm, VWD Injection Volume: 0.5 μl Sample Solvent: Acetonitrile/water Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro Extra column volume: ~4 μl Fenuron Fluomethuron Linuron Monuron Diuron Neburon This separation is an example of a rapid analysis of phenyl urea compounds on a HALO ES-CN column in under minutes. :

9 Application Note: 0-P Rapid Isocratic Separation of Fibrates on HALO C-8 Phase. Bezafibrate. Gemfibrozil. Fenofibrate t Column: 4.6 x 50 mm, HALO C-8 Part Number: Mobile Phase: 0/80-A/B A= 0.0 M Phosphate buffer, ph=.0 B=Acetonitrile Flow Rate:.5 ml/min. Pressure: 50 Bar Temperature: 45 C Detection: UV 0 nm, VWD Injection Volume: 0. μl Sample Solvent: 50/50-methanol/acetonitrile Response Time: 0.0 sec. Flow Cell:.5 μl semi-micro Extra column volume: ~4 μl Bezafibrate Fenofibrate Fibrates are a class of cholesterol lowering drugs that can be rapidly analyzed using Halo C-8 phase to obtain widely separated peaks in about 0 seconds. Gemfibrozil :

10 Application Note: 6-PT Separation of Peptides and Small Proteins on HALO Peptide ES-C G STRUCTUR. Gly Tyr. Val Tyr Val. Angiotensin ( 7) amide 4. Met Enk 5. Angiotensin ( 8) amide 6. Angiotensin II 7. Leu Enk 8. Ribonuclease A 9. Angiotensin ( ) (human) 0. Angiotensin ( ) (mouse). Porcine insulin Time (min.) Column: 4.6 x 50 mm, HALO Peptide ES-C8 Part Number: Mobile Phase: A= 90%(0.% TFA in water)/0% acetonitrile B= 0% (0.% TFA in water)/70% acetonitrile Gradient: 0% B to 87% B in minute Flow Rate: 5.0 ml/min. Pressure: 0 Bar Temperature: 60 C Detection: UV 0 nm, VWD Injection Volume:.0 μl Sample Solvent: Mobile Phase A Response Time: < 0. sec. Flow Cell: 5 μl semi-micro LC System: Quaternary Agilent 00 Gradient dwell volume is 0.88 ml DISCUSSION: This separation shows the utility of the HALO Fused-Core Peptide ES-C8 stationary phase for the separation of peptides by HPLC. An average pore size of about 60 Angstroms enhances the mass transfer of peptides and small proteins of up to a molecular weight of approximately 5 kd, depending on the molecular configuration. Also, the stationary phase is a sterically protected C8 bonded silane to increase resistance to low ph mobile phases and elevated temperatures (up to 00 C.) that are commonly used in the separation of many biological materials. HALO and Fused Core are registered trademarks of Advanced Materials Technology, Inc.