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Separation of Catecholamines |
 | Application Notes:
Primesep 200 separates catecholamines in less than 8 minutes. Tyrosine, dl-DOPA, dopamine, epinephrine are baseline resolved by a combination of reversed-phase, ion-exchange, and ion-exclusion mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
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 | Application Notes:
Primesep 200 separates catecholamines in the catecholamine pathway in 10 minutes. Phenylalanine, tyrosine, DOPA, dopamine, norepinephrine, and epinephrine are baseline resolved by a combination of reversed-phase, ion-exchange, and ion-exclusion mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
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Separation of Epinephrine Norepinephrine |
 | Application Notes:
Primesep 200 separates the catecholamines, norepinephrine and epinephrine, less than 8 minutes. Norepinephrine and epinephrine are baseline resolved by a combination of reversed-phase, and ion-exchange mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
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HPLC Separation of Compounds of Catecholamine Pathway |
 | Application Notes:
The catecholamine neurotransmitters are amino acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline resolved on a Primesep 200 column. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN), trifluoracetic acid (TFA) or ammonium formate and ultraviolet (UV) detection at 210 nm. Peak order and retention time can be changed by switching from TFA to ammonium formate in the mobile phase.
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HPLC Separation of Epinephrine and Epinephrine Sulfonate |
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HILIC Separation of Epinephrine and Epinephrine Sulfonate |
 | Application Notes:
Epinephrine (also referred to as adrenaline) is a hormone and neurotransmitter. It is a catecholamine, a sympathomimetic monoamine derived from the amino acids phenylalanine and tyrosine. This method can be used to determine and quantitate epinephrine and epinephrine sulfonate in biological fluids (urine, blood, serum) and drug formulations. Obelisc N columns are used to retain and separate epinephrine and epinephrine sulfonate in mixed-mode hydrophilic interaction chromatography. Epinephrine is retained by the combination of cation-exchange and HILIC mechanisms. Epinephrine sulfonate is retained by HILIC mechanism. Buffer concentration and pH, as well as the amount of acetonitrile, can be used to adjust retention of both compounds. Both compounds can be detected by UV, ELSD and LC/MS. Preparative separation is possible with volatile mobile phases (ammonium formate or ammonium acetate.
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Separation of Serotonin, Dopamine, and Related Compounds |
 | Application Notes:
Catecholamines are chemical compounds derived from the amino acid tyrosine containing catechol and amine groups. Some of them are biogenic amines. Retention of compounds of catecholamine pathway is achieved on Obelisc N column. All polar compounds are well retained by combination of HILIC and ion-exchange mechanisms. Obelisc N columns produce very good peak shapes for all analytes. Method is very sensitive to amount of ACN, buffer and buffer pH. Retention time changes with variation of main parameters. This method can be used for quantitation of biogenic amines and related compounds (homovanillic acid, dihydroxyphenyl acetic acid, serotonin, dopamine, epinephrine, hydroxytryptophan, epinephrine and DOPA) in urine, blood and other biological fluids. Further optimization of this HPLC method can be used during screening and validation. Amines and acids can be analyzed in the same run and retained by combination of polar organic mode, cation-exchange and anion-exchange modes. Various buffers within specified pH can be employed (ammonium formate, ammonium acetate, sodium phosphate, etc.).
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HPLC Separation of Neurotransmitters and Related Drugs |
 | Application Notes:
Epinephrine and norepinephrine (adrenaline and noradrenaline) are hormones and neurotransmitters. Epinephrine is synthesized from norepinephrine in a synthetic pathway shared by all catecholamines, including L-dopa, dopamine, norepinephrine, and epinephrine. Phenylephrine is used as a decongestant, available as an oral medicine or as a nasal spray. Phenylephrine is now the most common over-the-counter (OTC) decongestant. All three compounds are used in various drug compositions. Separation of epinephrine and norepinephrine is a challenging task due to polarity and close properties of two compounds. Epinephrine, norepinephrine and phenylephrine are separated in this method on Obelisc R mixed-mode HPLC columns. Method is very sensitive to variation of pH and pH adjustment can be used to achieve desired selectivity and retention time. Other catecholamines can be analyzed using this HPLC method. Method can be used as stability indicating or impurity profiling approach to analysis of neurotransmitters in drug formulation, blood, serum and urine.
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