Acetaminophen

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

Cold and cough remedies sold over the counter consist of several active and inactive ingredients, which are different in nature and resin differently in reversed-phase, ion-exchange, and HILIC chromatography. Wal-Dryl is a white label composition sold by Walgreens. Active Ingredients include Acetaminophen (325 mg), Diphenhydramine HCl (25 mg), Phenylephrine HCl (5 mg). Inactive Ingredients in this complex composition are Corn Starch, Croscarmellose Sodium, Crospovidone, FD&C Blue #1 Aluminum Lake, Hypromellose, Magnesium Stearate, Microcrystalline Cellulose, Polydextrose, Polyethylene Glycol, Povidone, Silica Gel, Stearic Acid, Titanium Dioxide, Triacetin. Mixed-mode HPLC columns allow retaining hydrophobic and hydrophilic, neutral and charged compounds in one run. Retention of polar basic compounds does not require the use of ion-pairing reagents (IP) to achieve controllable robust retention control. Order of elution for active and inactive ingredients in the complex mixture can be changed and adjusted with the pH of the mobile phase, as well as the amount of ACN and buffer. Coresep 100 retains and separates components of Wal-Dryl cold remedies with high efficiency and robustness. Acetaminophen or paracetamol is a relatively hydrophilic drug, which is used to treat pain and fever. Pseudoephedrine is a sympathomimetic medication in the phenylamine and amphetamine class of drugs. It is a stimulant. Pseudoephedrine is a part of numerous drug formulations used as cough suppressants. It is used as a precursor and starting material in the illicit production of methamphetamine and methcathinone. Pseudoephedrine is polar and basic in nature. Diphenhydramine is an antihistamine that reduces the effects of the natural chemical histamine in the body. Histamine can produce symptoms of sneezing, itching, watery eyes, and runny nose. Diphenhydramine is used to treat sneezing, runny nose, watery eyes, hives, skin rash, itching, and other cold or allergy symptoms. Diphenhydramine is hydrophobic and basic in nature compound. Excipients are hydrophilic and hydrophobic in nature and are added to improve taste and increase the stability of the formulation

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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:	Aromatic base, Aromatic compound, Drug, Stimulant
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Acetaminophen, Pseudoephedrine, Diphenhydramine

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

Hydrocodone/paracetamol, also known as hydrocodone/acetaminophen or hydrocodone/APAP is the combination of an opioid pain medication, hydrocodone, with paracetamol (acetaminophen). It is used as a prescription drug to relieve moderate to severe pain. It exists in tablet, elixir, and solution forms in various strengths for oral administration. Vicodin is Schedule II controlled substance. Hydrocodone diversion and recreational use has escalated in recent years due to its opioid effects. Norco was analyzed with isocratic conditions to provide fast, reliable and robust method for analysis of both components of the drug combination - hydrocodone and acetaminophen. Compounds are retained and separated on Heritage MA HPLC column in 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-exchange
Column Dimenstions:	4.6 x 150 mm
Mobile Phase:	5% ACN with 50 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:	Aromatic base, Aromatic compound, Controlled substance, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Hydrocodone, Acetaminophen

Codeine/paracetamol, also known as codeine/acetaminophen or hydrocodone/APAP is the combination of an opioid pain medication, codeine, with paracetamol (acetaminophen). It is used as a prescription drug to relieve moderate to severe pain. It exists in tablet, elixir, and solution forms in various strengths for oral administration. Vicodin is Schedule II controlled substance. Codeine diversion and recreational use has escalated in recent years due to its opioid effects. Vicodin was analyzed with isocratic conditions to provide fast, reliable and robust method for analysis of both components of the drug combination - codeine and acetaminophen. Compounds are retained and separated on Heritage MA HPLC column in 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 Coresep 100 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:	5% ACN with 50 mM AmAc pH 4
Detection:	UV 275 nm
Sample:	0.3 mg/ml
Injection:	3 uL
Flow rate:	1 mL/min

Analytes

Class of compounds:	Aromatic base, Aromatic compound, Controlled substance, Drug
Nature of compounds:	Basic, Hydrophilic, Hydrophobic, Neutral
Compounds:	Codeine, Acetaminophen

Core-shell mixed-mode columns offer unique selectivity which comes from the presence of two mechanisms - reversed-phase and ion-exchange. The alternative selectivity of mixed-mode columns can be used when order of elution or resolution between compounds needs improvement. Even for neutral compounds a different selectivity can be obtained. A method for common components of headache medications (acetaminophen, caffeine, and benzoate) was developed on our Coresep 100 column. Efficiency and peak shape are comparable to leading brand core-shell reversed-phase columns.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase
Column Dimenstions:	3.2 x 50 mm, 2.7 um, 90A
Mobile Phase:	20% ACN with 0.1% TFA
Detection:	UV 250 nm, ELSD
Sample:	0.3 mg/ml
Injection:	1 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Aromatic acid, Aromatic compound, Drug, Organic acid, Stimulant, Xanthine
Nature of compounds:	Acidic, Hydrophilic, Neutral
Compounds:	Acetaminophen, Caffeine, Benzoic acid