Proteins in Cream Formulation Identified Via Reverse-Phase High-Performance Liquid Chromatography

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Two scientists from Baskent University in Ankara, Turkey analyzed multiple substances in a cream formulation using reverse-phase high-performance liquid chromatography (RP–HPLC), according to a new study in Chromatographia (1).

For this experiment, the scientists focused on analyzing a select few substances: ruscogenin, neoruscogenin, trimebutine, and parabens, including methylparaben and propylparaben. Each of these proteins have different purposes in medicine; for example, ruscogenin is a steroid that helps in treating hemorrhoids, trimebutine is a spasmolytic agent used for treating irritable bowel syndrome (IBS), and parabens can act as endocrine system disruptors (2–4). Using a C18 column at 25°C for the separations, the mobile phase was made of acetonitrile and 20 mM phosphate buffer at pH 3.9, which was mixed in gradient elution mode using a binary pump. This method was validated and held to the requirements of the ICH Q2(R2) guideline.

The methods were deemed receptive towards all active substances and parabens. The limit of detection and limit of quantification values ranged from 0.02 to 0.45 µg mL–1 and from 0.06 to 1.35 µg mL–1, respectively. Additionally, the method’s intraday/interday accuracy (relative error ≤ 1.89) and precision (relative standard deviation ≤ 1.99) values were found in the limits, accounting for all compounds. According to validation studies, the method was found to be selective, sensitive, accurate, precise, and robust.



(1) Özen, G.; Nemutlu, E. Simultaneous Determination of Ruscogenin, Neoruscogenin, Trimebutin, and Parabens in Cream Formulation by Reverse Phase High Performance Liquid Chromatography. Chromatographia 2023, 86, 649–657. DOI:

(2) Ruscogenin. DrugBank. (accessed 2023-11-20)

(3) Trimebutine. DrugBank. (accessed 2023-11-20)

(4) Propylparaben. Environmental Working Group 2023. (accessed 2023-11-20)