Benzoic Acid

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Denatonium is the most bitter chemical compound known.
Denatonium benzoate is a substance that’s put into household, garden and automotive products to ensure that if a child or animal ever tried to swallow it, they would spit it straight out before they could be poisoned.

 

Condition 2

Column	Primesep SB, 4.6×150 mm, 5 µm, 100A
Mobile Phase	Gradient MeCN – 10-90%, 10 min
Buffer	Gradient H2SO4 – 0.9-0.1%, 10 min
Flow Rate	1.0 ml/min
Detection	UV, 275 nm

 

Description

Class of Compounds	Drug, The bitter chemical compound, Basic, Hydrophobic, Ionizable
Analyzing Compounds	Denatonium benzoate

 

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A general approach for analysis of various organic acids with MS detection in negative mode was developed using hydrogen-bonding stationary phase – SHARC 1. A highly sensitive method allows to analyze traces of organic acids in various matrices using ACN/MeOH/ammonia mobile phase. In hydrogen-bonding chromatography acetonitrile is a weaker solvent and alcohol is a stronger solvent. Various gradient and isocratic conditions can be used

Phthalic acid, phthalic acid isomers, and related products present in the production of phthalic acid were separated on the Primesep D column, based on reversed-phase and in-exchange mechanisms. Neutral, hydrophobic compounds of the phthalic acid production are retained by a reversed-phase mechanism, and phthalic acid and other acidic compounds are retained by a combination of reversed-phase and anion-exchange mechanisms. Resolution and selectivity of this separation can be modified by varying the amount of acetonitrile, buffer concentrations, and buffer pH. This method can be used for monitoring the production cycle of phthalic acid and related impurities.

Application Notes: Guaifenesin is common, over the counter expectorant. Guaifenesin contain no less than 98 percent and not more than 102 percent of the labeled amount of guaifenesin calculated on a dried basis, according to the USP methods. the The USP HPLC method for the analysis of guaifenesin was developed on our 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.

Application Columns: Legacy L1 C18 HPLC column

Application compounds: Guaifenesin, benzoic acid

Mobile phase: MeOH/H2O/AcOH 40:60:1.5

Detection technique: UV

Reference: USP 35- NF30

Application Notes: Acetametaphin, aspirin, and caffeine tablets contain not less than 90 percent and not more than 110 percent of the labeled amounts if acetametaphin, asprin, and caffeine according the USP methods. USP HPLC method for separation of acetaminophen, aspirin and caffeine was developed on Legacy L1 column according to US Pharmacopeia methodology. L1 classification is assigned to reversed-phase HPLC column contains C18 ligands. Support for the material is a 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: Acetaminophen, Aspirin, Caffeine, benzoic acid, and salicylic acid

Mobile phase: MeOH/H2O/AcOH 28/69/3

Detection technique: UV

Reference: USP30: NF35

Application Notes: Dopamine is a naturally occurring neurotransmitter found in the brain. Dopamine is a well studied compound because dopamine is an important neurotransmitter known to regulate many functions from pain response to behavior disorders. According to USP methods, dopamine hydrochloride contains not less than 98% and not more than 102% dopamine hydrochloride calculated on the dried basis. The USP HPLC method for the separation of hydrocortisone 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: Dopamine hydrochloride

Mobile phase: Water with 1% acetic and 5mM octanesulfonic acid/MeCN

Detection technique: UV

Reference: USP35: NF30

Mixed-mode HPLC columns allow to analyze compounds with drastically different properties in one run. Acidic, basic, and neutral compounds can be separated in one run using either isocratic or gradient conditions. In this application, neutral hydrophilic (uracil, phenol and hydroquinone), neutral hydrophobic (toluene), hydrophilic acidic (benzoic acid), hydrophilic basic (lutidine) and hydrophobic basic (amitriptyline) are separated using gradient of ACN. Neutral compounds are retained by reversed-phase mechanism, hydrophilic acidic compound become more hydrophobic at lower pH and retain by reversed-phase mechanism too. Basic compounds are retained by cation exchange mechanism, and hydrophobic basic compounds are retained by reversed-phase and cation-exchange mechanisms. All compounds are resolved within 17 minutes on a short column. Method can be applied to various polar and hydrophobic compounds, which can be separated on one column and in one run. Mixed-mode columns can operate in single or combination of several modes: reversed-phase, ion-exchange, ion-exclusion and HILIC. This mixed-mode HPLC column can be used as a general column for separation of wide range of compounds.

Primesep 100 and Primesep 200 columns can be used as a universal column for analysis of wide range of compounds. These mixed-mode reversed-phase ion-exchange HPLC columns can provide a valuable alternative to traditional reversed-phase column. Amines, amino acids, quaternary amines, and various zwitter-ions can be analyzed along with hydrophobic compounds and organic and inorganic counter-ions. In this application, 8 compounds with different hydrophobic, hydrophilic, basic and acidic properties are separated based on their properties. Primesep 100 column is a mixed-mode HPLC column with a C12 carbon chain and carboxylic acid on the surface with pKa of 1. Primesep 200 column is a mixed-mode HPLC column with a C12 carbon chain and carboxylic acid on the surface with pKa of 2. These columns can be used with 100% organic (ACN) and 100% aqueous mobile phases. This HPLC method can be adopted as a generic and robust approach for analysis of acidic, basic and neutral compounds within the same run.

Mixture of polar acidic and basic compounds is separated on a Primesep AB mixed-mode HPLC column. Dopamine and tyrosine are retained by combination of reversed-phase and cation-exchange mechanisms. Maleic acid is retained by anion-exchange mechanism, and benzoic acid is retained by reversed-phase mechanism. Primesep AB is a trimodal column with a C12 hydrophobic chain and cation-exchange and anion exchange groups on the surface. Method utilizes UV detection but can be used with other detection techniques (ELSD, LC/MS, Corona).

Primesep mixed-mode columns are designed to separate basic, acidic, neutral and zwitter-ionic compounds. Elution of these compounds is facilitated by acetonitrile and ions in the mobile phase. In all cases the retention time on mixed-mode columns is significantly longer than on traditional reverse-phase columns. Presence of cation-exchange mechanism in Primesep 100 column allows to achieve better retention and peak shape for analytes like phenylalanine and benzylamine. Various buffers can be used along with different detection techniques. This HPLC method can be adopted as a universal approach for analysis of basic, acidic and zwitter-ionic compounds in complex mixtures and formulations. Compounds can be monitored by UV, ELSD, CAD and LC/MS.

This application shows the effect of mobile phase composition on retention of acidic, basic and neutral compound. Retention of basic and acidic compounds is controlled by buffer pH and concentration, while retention of neutral compound is controlled by the amount of acetonitrile. Buffer pH changes not only ionization state of ionizable basic and acidic compounds, but ionization state of trimodal stationary phase Obelisc R. Available detection techniques include UV, ELSD, LC/MS, and Corona.

Excedrin is over-the-counter pain reliever containing acetaminophen, caffeine and aspirin as active ingredients of this drug composition. Acetaminophen (paracetamol) is used as analgesic and pain reliever. It is a neutral compound with low hydrophobicity. Aspirin or acetylsalicylic acid is used as analgesic and anti-inflammatory component of many OTC compositions. It is weakly acidic and slightly hydrophobic compound. Caffeine is xanthine alkaloid which is psychoactive stimulant drug. All four compounds are separated on mixed-mode Primesep 100 HPLC column with acetonitrile/water/TFA mobile phase. In this HPLC application compounds are retained by reversed phase mechanism. This HPLC method is short and robust.

Alginate is used in various pharmaceutical preparations. Chemically, it is a linear copolymer with homopolymeric blocks of (1-4)-linked ?-D-mannuronate (M) and its C-5 epimer ?-L-guluronate (G) residues, respectively, covalently linked together in different sequences or blocks. Alginic acid can be separated from benzoate, citric acid and saccharin by mixed-mode chromatography on Primesep C HPLC column. This method can be used to quantitate alginic acid, citric acid or saccharin in complex mixtures. Various detection technique can be used (UV, ELSD, LC/MS), based on mobile phase selection.

Obelisc N column are used for separation of weak and strong organic acids in mixed-mode HILIC. Benzoic and naphthalenesulfonic acids are retained based on polar interaction mode and anion-exchange mode. Order of elution and retention pattern can be changed by modifying mobile phase. PH of the mobile phase changes ionization state of stationary phase and analytes. Fast quantitation method for benzoic and naphthalenesulfonic acid can be developed using UV, ELSD or LC/MS detection. HPLC Method can be used for mixture of organic and inorganic strong and weak acids.

Mixed-mode chromatography allows separating, in single run, compounds with vastly different properties. A method for separation of amino acids (cysteine, methylcysteine, cystine and dimethylcysteine) in the presence of carboxylic acid (benzoic) and hydrophobic neutral compounds was developed on Primesep 100 mixed-mode column. At lower pH ionization of carboxylic acids is suppressed. Amino acids are retained as basic compound based on reverse phase and cation exchange mechanisms. Carboxylic acids are retained on this column based on weak reverse phase mechanisms. Neutral compounds are retained by reverse phase mechanism as on any other column. Retention time of basic, zwitter-ionic and hydrophobic compound can be adjusted by manipulation of mobile phase composition. ELSD, UV or LC/MS detection can be used based on the properties of analytes and mobile phase selection.

Separating basic, acidic and zwitterionic compounds in one run in reverse-phase HPLC can be very challenging. The methods might require the use of ion-pairing reagents and complex gradients that can make MS-compatibility difficult. Obelisc R column which has both positive and negative ion-pairs embedded in the stationary phase allows for fine tuning and separation of a wide range of compounds with different ionic properties. Acids, bases, amino acids and neutral compounds were separated isocratically in one run using a simple MS-compatible mobile phase of acetonitrile (ACN) and water with Ammonium Acetate (AmAc) buffer. Can also be UV detected at 250nm.

Primesep D separates organic acids such as fumaric, benzoic, phthalic, naphthoic, and maleic acids by a mixture of anion exchange and reversed phase. Retention times and elution order can be changed by adjusting the percentage of acetonitrile in the mobile. This can not be done by traditional ion-exchange and ion-exclusion chromatography. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoroacetic acid (TFA) and UV detection at 250 nm.

Primesep 100 separates a mixture of amino acids (tyrosine, phenylalanine), organic acids (benzoic acid, mandelic acid), amines (benzylamine, pyridine), and neutrals (benzonitrile, toluene) in one HPLC run by combining reversed-phase, cation-exchange, and polar interactions. The method is tunable and peak order can be changed significantly by adjusting acetonitrile and trifluoroacetic acid concentrations. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) and compatible with UV, mass spec (LC/MS) and evaporative light scattering (ELSD) detection.

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Parabens possess antibacterial and antifungal properties and are therefore widely used in pharmaceutical and cosmetic industries as preservatives in products. Parabens and benzoic acid can be baseline separated in a short time frame using Primesep B2 reverse-phase HPLC column with a simple mobile phase of water, acetonitrile (ACN, MeCN) and phosphoric acid of 0.1% as buffer. UV detection at 210nm.

Primesep B combines a hydrophobic, reversed-phase mechanism with ion exchange to separate the diacids, fumaric, benzoic, phthalic, naphthoic, and maleic acids. Changing the acetonitrile content of the mobile phase reverses the peak order for naphthoic and maleic acids. Primesep B combines reversed-phase and anion-exchange mechanism with a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) and UV detection at 250 nm.

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HPLC separation of carboxylic acids (nicotinic acid, benzoic acid and acetylbenzoic acid) on Primesep B2 column using an isocratic method by reverse-phase and anion-exchange mechanisms. The elution of carboxylic acids using the same isocratic method and mobile phase can be reversed by using a reverse-phase cation-exchange mechanisms of the Primesep 100 column. The mobile phase is water, acetonitrile (MeCN, ACN) and formic acid with a UV detector.

An HPLC method for the separation of active drug compounds on a Primesep 200 column. The retention of compounds is achieved through reverse-phase, cation exchange and hydrophobic interactions. Benzoic acid, hyoscyamine sulfate and phenyl salicylate are baseline separated using simple mobile phases of water, acetonitrile (MeCN, ACN) and trifluoroacetic acid (TFA) with a UV detector.

Primesep 100 separates a mixture of polar and nonpolar compounds in one analytical run. The amino acid cysteine; amino acid derivatives L-cystine, 2,2-dimethylcystine, and 2-methylcysteine; the polar acid benzoic acid; and the nonpolar neutral toluene are separated by a gradient using a combination of polar and hydrophobic interactions. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and sulfuric acid (H2SO4) with UV detection at 210 nm.