CAS Number | 59-42-7 |
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Molecular Formula | C9H13NO2 |
Molecular Weight | 167.208 |
InChI Key | SONNWYBIRXJNDC-VIFPVBQESA-N |
LogP | -0.310 |
Synonyms |
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Separation type: Bridge Ion Separation Technology, or BIST™ | ||||||||||||||
BIST™ Ionic Modifier Preparation | ||||||||||||||
High Performance Liquid Chromatography (HPLC) Method for Analysis of Phenylephrine, Epinephrine, Norepinephrine
Epinephrine and norepinephrine are key neurotransmitters and adrenal hormones naturally produced by the body to generate the fight-or-flight response, and synthesized by pharmaceutical companies to treat a host of issues, such as allergic reactions and low blood pressure. Phenylephrine is a related, synthetic compound made to treat low blood pressure, congestion, and hemorrhoids. Using SIELC’s newly introduced BIST™ method, these three similar neurotransmitters, which protonate in water, can be retained on a positively-charged anion-exchange BIST™ B+ column. There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged amine analytes to the positively-charged column surface and 2) a mobile phase consisting mostly of organic solvent to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Epinephrine, norepinephrine, and phenylephrine can be retained and UV detected at 200 nm.
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SIELC Technologies’ BIST™ Columns are a new and simple way to achieve many separations that are traditionally difficult or impossible to achieve with any other HPLC columns. When used with our BIST™ mobile phases, these ion exchange columns can generate very strong retention of analytes that have the same charge polarity as the stationary phase, unlocking new chromatography applications that were previously too difficult to achieve.
Select optionsSeparation type: Liquid Chromatography Mixed-mode
High Performance Liquid Chromatography (HPLC) Method for Analysis of Acetaminophen, Phenylephrine, and Ascorbic Acid
Acetaminophen (or paracetamol) is one of the most popular over-the-counter painkillers all over the world (in the US it is best known under the brand name Tylenol). Ascorbic acid (also known as Vitamin C) helps your body build up muscle, cartilage, and other important building blocks in your body. Phenylephrine is a common over-the-counter decongestant, but can also be used for pupil dilation and hemorrhoid treatment. While all of these compounds are available at your local pharmacy, Vitamin C intake can reduce the body’s ability to break down acetaminophen.
All three of these compounds can be measured at low UV. Using a Primsep S normal phase column and a mobile phase consisting of water and acetonitrile (MeCN) with an Ammonium acetate (AmAc) buffer, these three compounds can be separated and UV detected at 270 nm. Varying the buffer concentration changes the order in which phenylephrine and ascorbic acid are retained. This method is compatible with Mass Spectrometry.
Column | Primesep S2, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | Gradient MeCN – 97 to 70% |
Buffer | Gradient Ammonium Formate pH 3.0 – 10 to 50 mM |
Flow Rate | 1.0 ml/min |
Detection | UV 270 nm MS- compatible mobile phase |
Class of Compounds | Drug, Acid |
Analyzing Compounds | Acetaminophen, Phenylephrine, and Ascorbic Acid |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
Select optionsSeparation type: Liquid Chromatography Mixed-mode
High Performance Liquid Chromatography (HPLC) Method for Analysis of Acetaminophen, Phenylephrine, and Ascorbic Acid
Column | Primesep S, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O – 80/20% |
Buffer | Ammonium acetate pH 5.0 |
Flow Rate | 1.0 ml/min |
Detection | UV 270nm MS- compatible mobile phase |
Class of Compounds | Drug |
Analyzing Compounds | Acetaminophen, Phenylephrine, and Ascorbic Acid |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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Application Notes: Neurotransmitters are polar basic compounds. Several HPLC techniques are used to analyze these hydrophilic molecules. The most common techniques are reversed-phase with ion-pairing reagents chromatography, ion chromatography, HILIC, and mixed-mode chromatography. SIELC developed a new approach of analysis, which is based on hydrogen bond interactions. Hydrogen-bonding offers unique and alternative selectivity to traditional approaches like reversed-phase, HILIC, ion-exchange, and mixed-mode chromatography. Our methods are fully compatible with ELSD, LC/MS and prep chromatography.
Application Columns: SHARC 1, 3.2×100 mm, 5 um, 100A. To learn more about SHARC 1 columns click here. To order this column click here. To see more chromatographic separations check our web site.
Application compounds: Pseudoephedrine, norephedrine, phenylephrine and norphenylephrine
Detection technique: UV, LC/MS
Column | Sharc 1, 3.2×100 mm, 5 µm, 100A |
Mobile Phase | MeCN/MeOH |
Buffer | AmFm, Formic acid |
Flow Rate | 1.0 ml/min |
Detection | UV, 270 nm |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds | Pseudoephedrine, Norephedrine, Phenylephrine, Norphenylephrine |
The SHARC™ family of innovative columns are are the first commercially available columns with separation based primarily on hydrogen bonding. SHARC stands for Specific Hydrogen-bond Adsorption Resolution Column. Hydrogen bonding is an interaction (attraction) of bound hydrogen atom to the molecules with electronegative atoms such as oxygen, nitrogen, and fluorine.
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Phenylephrine is a hydrophilic basic compound. Phenylephrine is used primarily as a decongestant in cough composition. Tannic acid is poly-phenolic compound. Tannic acid is also used to produce tannate salts of certain antihistamins and anti-tussives to impart increased stability or slow release properties to the API (active pharmaceutical ingredient). Phenylephrine in the presence of tannic acid is analyzed by mixed-mode HPLC on a Primesep 100 column. Retention time for phenylephrine is controlled by the amount of acetonitrile and buffer concentration. Tannic acid elutes closer to void and not interferes with analysis of phenylephrine. Method can be used with UV detection.
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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Caffeine and phenylephrine are some of the components of various pain killer/fever reducers/cough compositions. In this application, both of these compounds are separated on Primesep 100 and Primesep 200 columns. Caffeine is retained by reversed-phase mechanisms and phenylephrine is retained by combination of reversed-phase and cation-exchange mechanisms. Retention time for caffeine is controlled by the amount of acetonitrile, while retention time of phenylephrine is controlled by amount of the acetonitrile, buffer concentration and buffer pH. The method can be used for analysis of components of pain, cough and cold medication in pharmaceutical production. Method is robust and reproducible and provides good retention and peak shape. Compounds are monitored by UV, ELSD, CAD or LC/MS. Analysis of active components on biofluids (urine, plasma, blood, etc) is possible with additional sample preparation (protein precipitation, SPE, etc.)
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
Select optionsThe Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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Acetaminophen is an over-the-counter pain and fever reducer, and a major component of cold and flue remedies. Phenylephrine is a decongestant. Guaifenesin is a mucolytic agent used to relieve respiratory difficulties. These three compounds of cough medication were separated by mixed-mode chromatography on a Primesep C HPLC column. Retention and order of elution for phenylephrine can be changed by buffer concentration and buffer pH. Method can be used for analysis of cough and cold composition during the production and in QC/QA environment. Method is compatible with LC/MS and can be used to analyze these components in biological fluids. This generic HPLC method is robust and reproducible.
Column | Primesep C, 3×150 mm, 5 µm, 100A |
Mobile Phase | MeCN |
Buffer | AmAc |
Flow Rate | 0.5 ml/min |
Detection | UV, 270 nm |
Class of Compounds |
Drug, Analgetic, Acid, Hydrophilic, Ionizable, |
Analyzing Compounds | Acetaminophen, Pseudoephedrine, Guaifenesin |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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Method for separation of components of cough medication shows how retention for acetaminophen, phenylephrine and guaifenesin is controlled. Order of elution for compounds can be changed based on composition of the mobile phase. The method is compatible with UV, ELSD and LC/MS and can be used for quantitation of drugs in formulation. Primesep C mixed-mode HPLC column is ideal candidate for analysis of cough compositions. Basic compounds are well retained in mixed-mode chromatography without use of ion-pairing reagents. Several companies validated this approach. The method is reproducible and robust and can be used in both production environments and R&D. This HPLC method can be adopted as general approach for analysis of basic compounds in various mixtures.
Column | Primesep C, 3×50 mm, 5 µm, 100A |
Mobile Phase | MeCN |
Buffer | AmAc |
Flow Rate | 0.5 ml/min |
Detection | UV, 270 nm |
Class of Compounds |
Drug, Analgetic, Acid, Hydrophilic, Ionizable, |
Analyzing Compounds | Acetaminophen, Pseudoephedrine, Guaifenesin |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
Select optionsEpinephrine 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.
Column | Obelisc R, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O |
Buffer | AmFm |
Flow Rate | 1.0 ml/min |
Detection | UV, 250 nm |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Hormone |
Analyzing Compounds | Phenylalanine, Norepinephrine, Epinephrine |
SIELC has developed the mixed-mode Obelisc™ columns to be the first commercially available columns with Liquid Separation Cell technology (LiSC™). This cost effective duo can replace multiple HPLC columns such as reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion- exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - Buffer concentration, Buffer pH, and Organic Modifier Concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.
Select optionsPrimesep C separates acetaminophen (paracetamol), phenylephrine, and diphenhydramine– common over-the-counter cold and flu medication ingredients such as Tylenol products—by either an acetonitrile or phosphoric acid gradient. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN), and phosphoric acid with ultraviolet (UV) detection at 210 nm.
Column | Primesep C, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | MeCN |
Buffer | H3PO4 |
Flow Rate | 1.0 ml/min |
Detection | UV, 210 nm |
Class of Compounds |
Drug, Analgetic, Acid, Hydrophilic, Ionizable, |
Analyzing Compounds | Acetaminophen, Pseudoephedrine, Diphenhydramine |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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