Dextromethorphan

There are several groups of compounds which are used as active and inactive ingredients in various over-the-counter drug compositions. These compounds are different in nature and can be organic and inorganic. Most of the active ingredients are either hydrophobic basic, hydrophilic basic or hydrophilic neutral compounds. Basic active ingredients of cough and cold remedies used as salts of organic and inorganic acids. The most commonly used hydrophobic active ingredients are doxylamine, pyrilamine, chlorpheniramine, brompheniramine, and dextromethorphan. Doxylamine is an antihistamine drug, used to relieve symptoms of allergy, hay fever, and the common cold. Doxylamine is used in combination with other active ingredients to provide effective nighttime relief from cough and cold symptoms. Pyrilamine is an antihistamine drug, which targetsH1 receptors as an inverse agonist. Brompheniramine is an antihistamine drug of propylamine nature. Chlorpheniramine is an antihistamine used to treat symptoms such as allergic rhinitis. Dextromethorphan is a medication mostly used as a cough suppressant. It is a morphine type of medication with sedative, dissociative, and stimulant properties. Succinic acid is a hydrophilic diacid that is used as a counterion. Maleic acid is hydrophilic diacid which is used as a counterion. Both compounds have very limited retention even in RP-AQ columns. Due to the strongly hydrophobic and basic properties of these compounds, they demonstrate a poor peak shape in traditional reversed-phase chromatography due to the overloading of residual silanol groups on the surface when these basic drugs are analyzed. In addition to this, hydrophilic counterions are not retained in RP because there is no mechanism of retention for these acidic hydrophilic compounds. Both problems were eliminated by using Heritage MA reversed-phase anion-exchange column. Column and method demonstrate high efficiency and good peak shape for acidic and basic analytes. It can be used to develop methods for various cough, cold, and congestion compositions without the use of ion-pairing reagents. Compounds are well retained by a combination of one or more mechanisms. The column is compatible with 100% ACN and 100% water as well as with most buffers with pH 1.5-7. Various additives to the mobile phase can be used based on the nature of samples and detection techniques.

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Conditions of Experiment

Column:	Heritage MA
Separation Modes:	reversed-phase, cation-exclusion, anion-exchange
Column Dimenstions:	4.6x150 mm, 3 um, 100A
Mobile Phase:	5% ACN with 30 mM AmFm pH 3
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Drug, Organic acid, Pyridine
Nature of compounds:	Acidic, Basic, Hydrophilic, Hydrophobic, Polar
Compounds:	Doxylamine, Pyrilamine, Succinic acid, Chlorpheniramine, Brompheniramine, Maleic acid, Dextromethorphan

Most of the over-the-counter cold and cough remedies contain a complex mixture of basic, acidic, neutral, hydrophilic, and hydrophobic compounds. Analysis of drugs and counterions usually require two methods - one for the active drug and related impurities and another one for analysis of counterions. The method can be simplified by switching to mixed-mode stationary phases. Mixed-mode stationary phases have ion-pairing reagent attached to the surface of silica gel. The presence of ionizable groups on the surface allows us to achieve 2-D separation on a single column. We have developed a reliable, robust high-efficiency method for analysis of active and inactive ingredients of Mucinex on Heritage MA mixed-mode column. The active ingredients of the formulation include guaifenesin and dextromethorphan. Guaifenesin is a hydrophilic organic compound that is used as a drug that is used to help to remove mucus from the mouth due to the common cold and flu. Dextromethorphan is a hydrophobic basic compound, which is used as an over-the-counter cough suppressant. Excipients are hydrophilic and hydrophobic in nature and are added to improve taste and increase the stability of the formulation. Heritage MA column shows exceptional peak shape and does not demonstrate poor peak shape for basic analytes like dextromethorphan. Heritage MA column can be used for analysis and validation of methods for separation of hydrophobic, hydrophobic basic, hydrophobic acidic and hydrophilic acidic compounds. There are several mechanisms available to be used with this column - reversed-phase, anion-exchange, cation-exclusion, and combination of the above.

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Conditions of Experiment

Column:	Heritage MA
Separation Modes:	reversed-phase, cation-exclusion, anion-exchange
Column Dimenstions:	4.6x150 mm, 3 um, 100A
Mobile Phase:	8% ACN with 30 mM NaH2PO4 pH 3.1
Detection:	UV 255
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug, Inorganic anion, Organic acid
Nature of compounds:	Acidic, Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Bromide ion, Dextromethorphan, Guaifenesin

Vick’s NyQuil Cold and Cough remedy temporarily relieves common cold/flu symptoms: cough due to minor throat & bronchial irritation, sore throat, headache, minor aches & pains - fever - runny nose & sneezing. The active ingredients include acetaminophen, chlorpheniramine, and dextromethorphan. Acetaminophen or paracetamol is a relatively hydrophilic drug, which is used to treat pain and fever. Dextromethorphan is a hydrophobic and basic compound, which serves as a cough suppressant in over-the-counter (OTC) medications. Chlorpheniramine hydrophobic and basic antihistamine drugs are used to treat symptoms of allergic conditions. All three active ingredients retain very differently, and usually, dextromethorphan and chlorpheniramine show a poor peak shape in RP chromatography due to residual silanol interactions. This problem does not exist on Coresep 100 mixed-mode column, where the polar ionizable group shields analytes from interacting with the residual silanols. Coresep 100 column demonstrates high efficiency and selectivity of separation. Methods for this drug composition are robust and reproducible and can be used in method development and validation. The column is compatible with 100% ACN and 100% water and can be used in a single or dual mode depending on the nature of the analyte and mobile phase composition. Very often order of elution can be changed by changing the amount of ACN, buffer pH, buffer concentration, and buffer nature.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Acetaminophen, Chlorpheniramine, Dextromethorphan

Mucinex Fast-Max Severe Congestion & Cough is a combination medicine used to treat cough, stuffy nose, and chest congestion caused by allergies, the common cold, or the flu. It is a complex mixture of several active and inactive ingredients. Active ingredients of this OTC drug composition are phenylephrine, guaifenesin, and dextromethorphan. Phenylephrine is a hydrophilic basic drug, which is primarily used as a decongestant, to dilate the pupil, to increase blood pressure, and to relieve hemorrhoids. Guaifenesin is a hydrophilic neutral compound which is added to cough and cold formulations to help with coughing thick mucus it is used in combination with dextromethorphan to alleviate symptoms related to cold, flu and congestion. Dextromethorphan is a hydrophobic and basic compound, which serves as a cough suppressant in over-the-counter (OTC) medications. Guaifenesin is retained by a pure reversed-phase mechanism, and dextromethorphan is retained by a combination of reversed-phase and cations-exchange mechanisms. Inactive ingredients consist of croscarmellose sodium, FD&C blue no. 2 aluminum lake, FD&C red no. 40 aluminum lake, methacrylic acid ethyl acrylate copolymer, mica, microcrystalline cellulose, polyethylene glycol 3350, polysorbate 80, polyvinyl alcohol, povidone K29/32, sodium bicarbonate, talc, titanium dioxide. Baseline separation of active and inactive ingredients was achieved on Coresep 100 mixed-mode column. Coresep 100 column provides good retention of basic compounds without the use of ion-pairing reagents and high pH. It retains compounds of different natures by a combination of reversed-phase, cation-exchange, and anion-exclusion mechanisms. The retention time of the robust separation is controlled by the amount of ACN, buffer concentration, and buffer. Various buffers and additives to the mobile phase can be used to control retention and be suitable with a chosen detection technique. This method and HPLC column can be used to achieve highly efficient, robust baseline separation of complex mixtures.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	A: 15% ACN with 0.02% H2SO4 B: 50% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Catecholamine, Drug, Neurotransmitter
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Phenylephrine, Guaifenesin, Dextromethorphan

Cough and cold drug composition are a complex mixture of compounds of different nature. Analysis of such drug composition might be challenging and require several methods of analysis including chromatography and titration. The main components of the medication are acetaminophen, pseudoephedrine and dextromethorphan. The liquid caps are used to temporarily relieve common cold and flu symptoms, like nasal congestion, cough due to minor throat & bronchial irritation, headache, minor aches & pains, fever and sore sore throat. Acetaminophen or paracetamol is a relatively hydrophilic drug, which is used to treat pain and fever. Dextromethorphan is a hydrophobic basic compound, which is used as an over-the-counter cough suppressant. Pseudoephedrine is a hydrophilic basic compound which is used as nasal and sinus decongestant or as a stimulant. Acetaminophen is retained by pure reversed-phase mechanisms and you can control retention time by the amount of ACN in the mobile phase. Dextromethorphan is retained by reversed-phase and cation-exchange mechanisms. The retention time of the dextromethorphan can be controlled by the amount of acetonitrile, buffer pH, and buffer concentration. Pseudoephedrine is mostly retained by the cation-exchange mechanism. Each mechanism of interaction can be controlled separately resulting in unprecedented selectivity of mixed-mode columns. Core-shell particles of the Coresep 100 HPLC column ensure high efficiency of separation associated with core-shell particles. Various buffers and acids can be used to accommodate specific detection techniques like UV, ELSD, CAD, and mass spectrometry. Coresep 100 columns can be used to analyze basic compounds without ion-pairing reagent in the mobile phase. Hydrophobic neutral, hydrophobic basic, hydrophilic basic, hydrophobic basic and weak carboxylic acids can be analyzed with Coresep 100 column. Hydrophilic acids might have limited retention and require a different column (Coresep SB, Heritage MA). Mixed-mode HPLC columns provide 2-D selectivity for the retention of hydrophobic and hydrophilic ionizable compounds.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Acetaminophen, Dextromethorphan

Robitussin Severe Cough, Cold & Flu Formulation is a complex mixture of organic and inorganic compounds. The main active drug ingredients are acetaminophen, phenylephrine, guaifenesin, and dextromethorphan. Inactive ingredients include anhydrous citric acid, edetate disodium, FD&C red no. 40, glycerin, menthol, natural and artificial flavors, polyethylene glycol, propyl gallate, propylene glycol, purified water, sodium benzoate, sodium citrate, sorbitol solution, sucralose, triacetin, xanthan gum. Acetaminophen or paracetamol is a relatively hydrophilic drug, which is used to treat pain and fever. Phenylephrine is a hydrophilic basic drug, which is primarily used as a decongestant, to dilate the pupil, to increase blood pressure, and to relieve hemorrhoids. Guaifenesin is a hydrophilic organic compound which is used as a drug that is used to help to remove mucus from the mouth due to the common cold and flu. Dextromethorphan is a hydrophobic basic compound, which is used as an over-the-counter cough suppressant. Excipients are hydrophilic and hydrophobic in nature and are added to improve taste and increase stability of the formulation. Several complex methods are required to fully analyze this drug composition. We have developed a simple UV method using Coresep 100 mixed-mode column. Coresep 100 column is a reversed-phase cation-exchange column, based on 2.7 um core-shell articles. It is designed to retain various analytes without the use of ion-pairing reagents. This approach makes these columns fully compatible with mass spec detection. The method for analysis of this drug formulation is reproducible and robust and provides baseline separation of hydrophobic basic, hydrophilic basic, neutral, hydrophobic, and hydrophobic acidic compounds. Hydrophilic acidic compounds might not be retained. The column is compatible with 100% water and 100% ACN and totally resistant to phase collapse, due to polar-embedded ionizable groups on the surface of core-shell particles. Mixed-mode HPLC columns provide 2-D selectivity for the retention of hydrophobic and hydrophilic ionizable compounds.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.03% H2SO4 B: 55% ACN with 0.25% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Catecholamine, Drug, Neurotransmitter
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Acetaminophen, Phenylephrine, Guaifenesin, Dextromethorphan

Robitussin Severe Cold & Sore is a drug combination consisting of active ingredients and excipients. Active ingredients include acetaminophen and dextromethorphan. Excipients or inactive ingredients include anhydrous citric acid, edetate disodium, FD&C red no. 40, glycerin, menthol, natural and artificial flavors, polyethylene glycol, propyl gallate, propylene glycol, purified water, sodium benzoate, sodium citrate, sorbitol solution, sucralose, triacetin, xanthan gum. Complex composition like this can be successfully analyzed in isocratic or gradient mode on Coresep 100 mixed-mode core-shell reversed-phase cation-exchange column. Compounds are retained and separated by reversed-phase, cation-exchange and anion-exclusion mechanisms. Retention time is controlled by the amount of ACN, buffer concentration and buffer pH. Method is robust and reproducible and can be used for analysis of other over-the-counter (OTC) formulations. Depending on the properties of the compounds and detection technique, other buffers and additives can be used to achieve the desired separation, selectivity, and resolution between critical compounds. Compounds are well retained without the use of ion-pairing reagent. The column is compatible with 100% water and 100% ACN. Other drug formulation can be analyzed on mixed-mode columns with perfect peak shape, retention control, and robustness. Coresep 100 columns can be used with any of the common detection techniques (UV, ELSD, RI, mass-spec, CAD). Mixed-mode HPLC columns provide 2-D selectivity for the retention of hydrophobic and hydrophilic ionizable compounds.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Acetaminophen, Dextromethorphan

Alka-Seltzer Plus MAX Strength Severe Sinus is a complex mixture of active ingredients and excipients. It is used in the treatment of cough, stuffy nose, body aches, headache, and a sore throat caused by common cold and flu. The active ingredients include acetaminophen, phenylephrine, doxylamine, and dextromethorphan. Inactive ingredients and excipients include D&C yellow #10, FD&C blue #1, gelatin, glycerin, polyethylene glycol, povidone, propylene glycol, purified water, shellac, sodium hydroxide, sorbitol sorbitan solution, titanium dioxide. These components are different in nature. Acetaminophen is a neutral relatively hydrophobic compound, doxylamine is a basic hydrophobic compound. Dextromethorphan is a hydrophobic basic compound. Phenylephrine is a hydrophilic basic compound. Excipients are polar in nature with some acidic and basic additives. Analysis of such complex mixtures is a difficult task and requires a different methodology. We have developed a robust and reproducible method for the analysis of these drugs, impurities, and excipients. Baseline HPLC separation is achieved on Corespe 100 mixed-mode reversed-phase cation-exchange core-shell column. Column demonstrates good peak shape and retention control. Unique selectivity of mixed-mode columns allows controlling the retention of compounds of different nature independently by changing mobile phase composition. The selectivity of the separation controlled by the amount of ACn, buffer concentration, and buffer pH. Various buffers and acids can be used as additives to the mobile phase. The choice of the buffer depends on the detection technique and properties of the column and analytes. Core-shell mixed-mode columns offer unprecedented selectivity associated with multi-mode interactions and high efficiency of core-shell particles. Compounds are well retained without the use of ion-pairing reagent. The column is compatible with 100% water and 100% ACN. Other drug formulation can be analyzed on mixed-mode columns with perfect peak shape, retention control, and robustness. Coresep 100 columns can be used with any of the common detection techniques (UV, ELSD, RI, mass-spec, CAD).

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Catecholamine, Drug, Neurotransmitter
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Acetaminophen, Phenylephrine, Doxylamine, Dextromethorphan

Tylenol Severe Cold and Flu formulation includes active ingredients as well as excipients. Active components include acetaminophen, phenylephrine, guaifenesin, and dextromethorphan. Inactive ingredients include carnauba wax, croscarmellose sodium, D&C yellow no. 10 aluminum lake, flavor, hydroxypropyl cellulose, hypromellose, magnesium stearate, microcrystalline cellulose, polyethylene glycol, pregelatinized starch, sucralose, titanium dioxide. The complex mixture of hydrophobic, hydrophilic, basic, acidic, and neutral components of Tylenol medication was fully separated on Coresep 100 mixed-mode core-shell reversed-phase cation-exchange column. The method is robust and reproducible and can be used for the analysis of drugs and drug compositions in various sample matrices. Various buffers and additives to the mobile phase can be used depending on the detection technique. The retention time of active ingredients, impurities, and excipients is controlled by the amount of acetonitrile (ACN), buffer pH, and buffer concentration. Core-shell columns provide very high efficiency of separation and mixed-mode provides unique selectivity and ability to retain hydrophilic compounds without the use of ion-pairing reagents. Mixed-mode columns are compatible with 100% ACN and 100% water due to the presence of polar and ionizable groups on the surface of silica gel. The column can be effectively used in isocratic and gradient runs to achieve baseline resolution between various active and inactive ingredients.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.03% H2SO4 B: 70% ACN with 0.25% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Catecholamine, Drug, Neurotransmitter
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Acetaminophen, Phenylephrine, Guaifenesin, Dextromethorphan

Tussin Cough and Cold formulation consists of medication to relieve cough caused by the common cold, bronchitis, or other respiratory issues. The active ingredients include guaifenesin and dextromethorphan. Guaifenesin belongs to a class of drugs known as expectorants. It helps to deal with mucus, clearing congestion and making breathing easier. Dextromethorphan belongs to a class of drugs known as a cough suppressant, it works on the part of the brain to reduce the urge to cough. Inactive ingredients include anhydrous citric acid, FD&C red 40, flavor, glucose, glycerin, high fructose corn syrup, menthol, purified water, saccharin sodium, sodium benzoate. Active ingredients and excipients were separated on Coresep 100 mixed-mode reversed-phase cation-exchange column with a simple gradient. Baseline separation was achieved under 10 minutes and showed perfect peak shape and robustness. The method uses ACN/water/sulfuric acid to elute all compounds. Other buffers and additives can be used to retain and separate hydrophobic, hydrophilic, neutral, and basic compounds. Other buffers and additives can be used to replace sulfuric acid and accommodate detection techniques where the volatile mobile phase is required (ELSD, CAD, mass-spectrometry). The retention time of active ingredients, impurities, and excipients is controlled by the amount of acetonitrile (ACN), buffer pH, and buffer concentration. Core-shell columns provide very high efficiency of separation and mixed-mode provides unique selectivity and ability to retain hydrophilic compounds without the use of ion-pairing reagents.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Guaifenesin, Dextromethorphan

Dimetapp Cold & Cough for kids is a liquid medicine containing three active ingredients: a nasal decongestant (Phenylephrine HCI), an antihistamine (Brompheniramine maleate) and a cough suppressant (Dextromethorphan HBr). There are multiple excipients in the formulation for the flavor, thickness, and preservation. The nasal decongestant works to temporarily restore free breathing through the nose to help clear a stuffy nose. The antihistamine/cough suppressant in this liquid cough medicine is tough on common upper respiratory symptoms like runny nose, sneezing, itchy, watery eyes, and itching of the nose or throat, as well as coughing due to minor throat irritation. Phenylephrine is a hydrophilic basic compound with poor retention in reversed-phase chromatography. Brompheniramine is a hydrophobic basic compound with strong retention and poor peak shape on traditional reversed-phase columns. Dextromethorphan is a hydrophobic basic compound with strong retention and poor peak shape when reversed-phase columns are used. We have developed a simple but robust method for analysis of active ingredients and excipients of Dimetapp formulation. All active compounds and excipients are retained and separated by several mechanisms - reversed-phase, cation-exchange, and anion-exclusion. Inactive ingredients in the formulation include Anhydrous Citric Acid, Artificial Flavor, Fd&C Blue No. 1, Fd&C Red No. 40, Glycerin, Propylene Glycol, Purified Water, Sodium Benzoate, Sodium Citrate, Sorbitol Solution, Sucralose. The retention of compounds is adjusted by changing the amount of ACN, buffer pH, and buffer concentration. The method can be applied to the analysis of drugs in biofluids. Most common detection techniques can be used (UV, ELSD, mass-spec, etc.). Core-shell columns provide very high efficiency of separation and mixed-mode provides unique selectivity and ability to retain hydrophilic compounds without the use of ion-pairing reagents.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	"A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold "
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Catecholamine, Drug, Neurotransmitter
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Polar
Compounds:	Phenylephrine, Dextromethorphan, Brompheniramine

Cough, cold and flu medications are a complex mixture of active ingredients and excipients. It usually contains hydrophilic and hydrophobic drugs, counterions, other organic and inorganic additives. Analysis of such complex mixtures are a challenging task. Analysis is usually done with several methods, including reversed-phase and ion-exchange chromatography. Active ingredients of Wal-Tussin DM MAX (guaifenesin and dextromethorphan and excipients were separated on Coresep 100 mixed-mode reversed-phase and cation-exchange HPLC column. Guaifenesin is a hydrophilic neutral compound which is added to cough and cold formulations to help with coughing thick mucus it is used in combination with dextromethorphan to alleviate symptoms related to cold, flu and congestion. Dextromethorphan is a hydrophobic and basic compound, which serves as a cough suppressant in over-the-counter (OTC) medications. Guaifenesin is retained by a pure reversed-phase mechanism, and dextromethorphan is retained by a combination of reversed-phase and cations-exchange mechanisms. This HPLC method demonstrates baseline separation and robustness in analysis of cough and cold formulations. Coresep 100 columns can be successfully used for analysis of various compounds, drug compositions and formulations. The column is compatible with 100% water and 100% ACN and can be used in isocratic or various gradient modes. The retention of compounds is adjusted by changing the amount of ACN, buffer pH, and buffer concentration. The method can be applied to the analysis of drugs in biofluids. Most common detection techniques can be used (UV, ELSD, mass-spec, etc.). Core-shell columns provide very high efficiency of separation and mixed-mode provides unique selectivity and ability to retain hydrophilic compounds without the use of ion-pairing reagents.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6x150 mm, 2.7 um, 90A
Mobile Phase:	A: 10% ACN with 0.02% H2SO4 B: 70% ACN with 0.2% H2SO4 from 100% A to 100% B in 8 min, 4 min hold
Detection:	UV 275
Sample:	various
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Aromatic compound, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral, Polar
Compounds:	Guaifenesin, Dextromethorphan

Dextromethorphan is a drug of the morphinan class with sedative, dissociative, and stimulant properties (at higher doses). It is a cough suppressant in many over-the-counter (OTC) cold and cough medicines including Benylin DM, Mucinex DM, Robitussin, NyQuil, Dimetapp, Vicks, Coricidin, Delsym, TheraFlu, Cheracol D, and others. Dextromethorphan has also found numerous other uses in medicine, ranging from pain relief (as either the primary analgesic or an opioid potentiator) over psychological applications to the treatment of addiction. Dextromethorphan and related impurities were analyzed by mixed-mode chromatography on Heritage MA HPLC column. Compounds are separated by reversed-phase and cation-exclusion mechanisms. Retention time, selectivity, and resolution between peaks can be adjusted by modification of amount of ACN, buffer and buffer pH. Various mobile phases can be used with Heritage MA column to accommodate different detection techniques (UV, MS, CAD, ELSD detection).

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Conditions of Experiment

Column:	Heritage MA
Separation Modes:	reversed-phase and cation-exclusion
Column Dimenstions:	4.6 x 150 mm
Mobile Phase:	15% ACN with 30 mM AmAc pH 4
Detection:	UV 215 nm
Sample:	0.3 mg/ml
Injection:	3 uL
Flow rate:	1 mL/min

Analytes

Class of compounds:	Amines, Aromatic base, Drug
Nature of compounds:	Basic, Hydrophobic
Compounds:	Dextromethorphan

Dextromethorphan and bromide counter-ion were analyzed by mixed-mode HPLC on core-shell reversed-phase anion exchange column. Dextromethorphan is retained by strong hydrophobic interaction and the bromide ion is retained by an anion-exchange interaction. Order of elution for basic drug and it’s acidic counter-ion can be changed by modifying the amount of ACN, buffer pH, and buffer concentration. The Coresep SB column can be used for analysis of hydrophobic basic drugs and acidic counter-ions in one run without a ion-pairing reagent. Compounds can be monitored by all common detection techniques.

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Conditions of Experiment

Column:	Coresep SB
Separation Modes:	reversed-phase, anion-exchange and cation-exclusion
Column Dimenstions:	3.2 x 100 mm, 2.7 um, 90A
Mobile Phase:	20% ACN with 0.1% or 0.2% TFA
Detection:	UV 255 nm
Sample:	0.3 mg/ml
Injection:	3 uL
Flow rate:	0.6 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Drug
Nature of compounds:	Basic, Hydrophobic
Compounds:	Dextromethorphan

Three hydrophobic drugs were analyzed with excellent peak shape on the core-shell mixed-mode column. Poor peak shape is very often observed when hydrophobic basic drugs, like dextromethorphan and trimipramine, are analyzed by traditional reversed-phase chromatography. This poor peak shape is caused by the interaction with residual silanol groups on the surface of RP columns. Since the amounts of these groups are not great, easy overloading and tailing occur. In case of mixed-mode chromatography, the polar basic group on the surface of silica gel provides a shielding effect, preventing the interaction of basic groups of analyte and acidic residual silanols. The mixed-mode core-shell column also has a high capacity towards retention of acidic drugs, which contributes to the good peak shape and retention. Method and column can be used to develop robust and reproducible methods for a hydrophobic basic, hydrophilic acidic and hydrophobic acidic compounds and counter-ions.

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Conditions of Experiment

Column:	Coresep SB
Separation Modes:	reversed-phase and cation-exclusion
Column Dimenstions:	3.2 x 100 mm, 2.7 um, 90A
Mobile Phase:	25% ACN with 120 mM of AmFm pH 3
Detection:	UV 250 nm, ELSD
Sample:	0.3 mg/ml
Injection:	1 uL
Flow rate:	0.8 ml/min

Analytes

Class of compounds:	Amines, Aromatic base, Drug
Nature of compounds:	Basic, Hydrophobic
Compounds:	Dextromethorphan, Verapamil, Trimipramine