Propylparaben

Parabens are used as preservatives in food, beverages and drug formulations. Methyl, ethyl and propyl esters of 4-hydroxybenzoic acid are used in many formulations, mixtures and compositions as preservatives. Parabens are neutral in nature, and they are retained on Heritage HA mixed-mode column by pure reversed-phase mechanism. The method can be used in the analysis of parabens and other neutral or acidic preservatives by LC-UV and LC-MS. The retention time is controlled by the amount of organic component in the mobile phase. This method is robust and reproducible.

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Conditions of Experiment

Column:	Heritage MA
Separation Modes:	reversed-phase
Column Dimenstions:	4.6x150 mm, 3 um, 100A
Mobile Phase:	60% ACN with 10 mM Am Ac ph 4
Detection:	UV 255 nm
Sample:	0.2 mg/ml
Injection:	3 ul
Flow rate:	1 ml/min

Analytes

Class of compounds:	Aromatic compound, Organic compound, Preservative
Nature of compounds:	Hydrophobic, Neutral
Compounds:	Methylparaben, Ethylparaben, Propylparaben

Parabens are used as preservatives in food and drugs. Methyl, ethyl and propyl esters of 4-hydroxybenzoic acid are used in many formulations, mixtures and compositions. Parabens are neutral and hydrophobic in nature, and they are retained on a Heritage HA mixed-mode column by pure reversed-phase mechanism. The method can be used in the analysis of parabens and other neutral or acidic preservatives by LC-UV and LC-MS. The retention time is controlled by the amount of organic component of the mobile phase. This method is robust and reproducible.

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Conditions of Experiment

Column:	Heritage MA
Separation Modes:	reversed-phase
Column Dimenstions:	4.6x150 mm, 3 um, 100A
Mobile Phase:	60% ACN with 10 mM Am Ac ph 4
Detection:	UV 255 nm
Sample:	0.2 mg/ml
Injection:	3 ul
Flow rate:	1 ml/min

Analytes

Class of compounds:	Aromatic compound, Organic compound, Preservative
Nature of compounds:	Hydrophobic, Neutral
Compounds:	Methylparaben, Ethylparaben, Propylparaben

Seven compounds with different properties were used to study an effect of different acids in the mobile phase on retention of neutral, acidic, basic and zwitterionic compounds on core-shell mixed-mode Coresep 100 column. Retention of neutral compounds (propylparaben and benzoic acid to some extent) are not affected by change of the acid. Basic compounds (norphenylephrine, pyridine, 2-aminopyridine) respond the most to change of the strength of acidic additive. Several folds increase in retention time was observed when sulfuric acid was replaced with formic acid in the mobile phase. Effect of acid on retention time of zwitterionic amino acids was less pronounced but still provided excellent retention and peak shape for zwitterionic compounds (5-aminosalicylic acid, phenylalanine). This methods and Coresep 100 column can be used to retain and separate other acidic, basic, neutral and zwitterionic compounds without the use of ion-pairing reagent in the mobile phase. Various detection techniques can be used based on the properties of the mobile phase and analytes. Methods are compatible with mall major detection techniques 9UV, ELSD, CAD, RI, MS). Core-shell mixed-mode columns provide great combination of unique selectivity, speed and efficiency.

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Conditions of Experiment

Column:	Coresep 100
Separation Modes:	reversed-phase, cation-exchange
Column Dimenstions:	4.6 x 150 mm, 2.7 um, 90A
Mobile Phase:	30% ACN with various acids
Detection:	UV 255 nm
Sample:	0.3 mg/ml
Injection:	3 uL
Flow rate:	1 mL/min

Analytes

Class of compounds:	Amines, Amino acid, Aromatic acid, Aromatic base, Aromatic compound, Catecholamine, Drug, Isomer, Neurotransmitter, Organic acid, Preservative, Pyridine, Supplement
Nature of compounds:	Acidic, Basic, Hydrophilic, Hydrophobic, Neutral, Polar, Zwitterionic
Compounds:	5-Aminosalicylic acid, Norphenylephrine, Phenylalanine, Pyridine, Benzoic acid, 2-Aminopyridine, Propylparaben

United States Pharmacopeia establishes written procedures for analysis of drugs, food ingredients, dietary supplement products and other ingredients. These standard procedures are used by pharmaceutical, chemical and other companies to ensure identity and purity of various compounds. Propylparaben, along with methylparaben and ethylparaben, is a preservative used in water-based cosmetics and food. Parabens are hydrophobic neutral compounds that can be easily retained on a Heritage C18 HPLC column. Analysis and column are robust and reproducible and can be used in determination of parabens in various samples. Retention time is controlled by amount of ACN or MeOH in the mobile phase. Buffers and additives can be chosen to accommodate different detection techniques.

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Conditions of Experiment

Column:	Heritage С18
Separation Modes:	reversed-phase
Column Dimenstions:	4.6x150 mm, 5 um, 100A
Mobile Phase:	50% MeCN with 20 mM NaH2PO4 pH 2.5
Detection:	UV 254 nm
Sample:	0.3 mg/ml
Injection:	5 uL
Flow rate:	1 mL/min

Analytes

Class of compounds:	Aromatic compound, Preservative
Nature of compounds:	Hydrophobic, Neutral
Compounds:	Propylparaben