Epinephrine

Applications:

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HPLC Separation of Catecholamines

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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Dopamine
Epinephrine

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HPLC Analysis of the Catecholamine Pathway

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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DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine

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Separation of Epinephrine Norepinephrine

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 trifluoroacetic acid (TFA) with UV detection at 210 nm.

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Epinephrine
Norepinephrine

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HPLC Separation of Compounds of Catecholamine Pathway

The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a Primesep 200 column with UV-transparent phosphate buffer. This method can be used for the analysis of catecholamines and related impurities in various matrices. The peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate the desired detection technique. Primesep 200 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitterionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and the nature of the analytes. The column is compatible with LC/MS and does not require the use of ion-pairing reagents.

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DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine

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HPLC Separation of Epinephrine and Epinephrine Sulfonate

Epinephrine and epinephrine sulfonate are separated on combination of Primesep 100 and Primesep SB HPLC columns. Primesep 100 column retains epinephrine by combination of reversed-phase and cation-exchange mechanisms. Epinephrine sulfonate is retained by weak reversed-phase mechanism. Due to strong zwitter-ionic nature of epinephrine sulfonate, ionic interaction is not available for this molecule. Compounds can be monitored by UV, ELSD, CAD or LC/MS.

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Epinephrine
Epinephrine Sulfonate

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HILIC Separation of Epinephrine and Epinephrine Sulfonate

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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Epinephrine
Epinephrine Sulfonate

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Separation of Serotonin, Dopamine, and Related Compounds

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 the 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. The method is very sensitive to amount of ACN, buffer and buffer pH. The retention time changes with variation of the 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 a 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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DOPA (3,4-dihydroxy-L-phenylalanine)
DOPAC (Dihydroxyphenylacetic Acid)
Dopamine
Epinephrine
Homovanillic Acid
Hydroxytryptophan
Norepinephrine
Serotonin

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HPLC Separation of Neurotransmitters and Related Drugs

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. The 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. The method can be used as a stability indicating or a impurity profiling approach to the analysis of neurotransmitters in drug formulation, blood, serum and urine.

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Epinephrine
Norepinephrine
Phenylephrine

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HPLC Separation of Epinephrine and Related Impurities

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. Epinephrine is polar basic compounds and it is retained on mixed-mode cation exchange columns without ion-pairing reagent. Epinephrine is retained by weak reversed phase and strong cation-exchange mechanisms. Formulations for epinephrine might have citric and ascorbic acid and EDTA. These three compounds are not retained by cation-exchange or reversed-phase mechanisms and elute in the void. Current HPLC method can be used for quantitation of epinephrine and recepinephrine (recemate) in various compositions. Epinephrine and related impurity can be monitored by UV, ELCD/MS, ELSD or Corona CAD. Corresponding buffer is required for each detection technique.

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Epinephrine
Racepinephrine

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HPLC Separation of Catecholamines on Primesep 100 Column with Phosphate Buffers

The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a Primesep 100 column with UV-transparent phosphate buffer. This method can be used for analysis of catecholamines and related impurities in various matrices. Peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate desired detection technique. Primesep 100 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitter-ionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and nature of analytes. Column is compatible with LC/MS and does not require use of ion-pairing reagents.

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DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Norepinephrine
Phenylalanine
Tyrosine

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HPLC Separation of Dopamine and Epinephrine in HILIC and Cation-Exchange Modes

Dopamine and epinephrine were separated on a Primesep S2 HILIC mixed-mode column. Dopamine and epinephrine are retained by HILIC and cation-exchange mechanisms. Retention time is controlled by the amount of ACN, buffer and buffer pH. Method is compatible with LC/MS and can be used for analysis of polar metabolites in biofluids.

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Dopamine
Epinephrine

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USP Methods for the Analysis of Epinephrine Using the Legacy L1 Column

 

Application Notes: Epinephrine is a synthetic adrenaline used to treat cardiac arrest and anaphylaxis. Phenylephrine is a decongestant and is often used instead of pseudoephedrine. According to the USP methods epinephrine contains no less than 97% and no more than 100.5 percent of epinephrine calculated on a dried basis. The USP HPLC method for the separation of phenylephrine and epinephrine was developed on Legacy L1 column according to the US Pharmacopeia methodology. L1 classification is assigned to reversed-phase HPLC column containing C18 ligand. Support for the material is spherical silica gel with particles size 3-10 um and pore size of 100-120A. Resolution between critical pairs corresponds to rules and specifications of USP.

Application Columns: Legacy L1 C18 HPLC column

Application compounds: Epinephrine and Phenylphrine
Mobile phase: Water/MeOH (50/50) with 1% 1-Octanesulfonic acid adjust to pH 3.0 with H3PO4

Detection technique: UV

Reference: USP35: NF30

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Epinephrine

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HPLC Separation of Epinephrine on Legacy L1 Column

Epinephrine was analyzed according to the US Pharmacopeia method on traditional reversed-phase HPLC columns, classified as a L1 type of column according to USP classification. Legacy L1 column can be used for the analysis of drug substances in various formulations. Legacy L1 can be used in new HPLC methods as well as in validated methods which require experiments based on USP manuscript. The mobile phase can be modified to accommodate other detection techniques, like LC/MS.

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Epinephrine

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HPLC Analysis of Epinephrine and Epinephrine Sulfonate on Trimodal Column

 

Epinephrine and epinephrine sulfonate were analyzed using Primesep AB, a reverse-phase column with both acidic and basic embedded ion-pairing groups. This method is LC/MS compatible.

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Epinephrine
Epinephrine Sulfonate

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Generic Screening Method for Complex Mixtures

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2,3-Dihydroxybenzoic Acid
2,6-Lutidine
Amitriptyline
Benzoic Acid
Benzylamine
DOPA (3,4-dihydroxy-L-phenylalanine)
Epinephrine
Ethyl Paraben
Homovanillic Acid
Hydroxytryptophan
Methylparaben
Phenol
Toluene
Tryptophan
Uracil

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HPLC Determination of Epinephrine in Human Plasma Sample

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FlipLC™ is an alternative method to avoid the interference of most of the contaminants by the use of an isolation column and a high pressure switching valve before the separation column. This method allows sample cleaning and analyte separation in one automated process. The isolation column and the separation column should have orthogonal retention characteristics to operate efficiently in this setup. Mixed-mode columns with reverse phase and ion-exchange characteristics were used in this analysis.

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Epinephrine
Epinephrine bitartrate

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