Rapid HPLC Method for Determination of Parachloroaniline in Chlorhexidine Antiseptic Agent in Mouthrinses, Ophthalmic and Skin Solution

Abstract

We described a simple and rapid method to quantify simultaneously chlorhexidine (CHD) and its major metabolite, para Chloroaniline (pCA) by HPLC with UV detection without the additional need of mobile-phase amine modifiers or ion-pairing reagents, with good resolution between pCA and CHD, symmetry peak of the compound and short run time. HPLC-UV analyses were performed using a Dionex® Summit liquid chromatograph (Dionex Corp, Sunnyvale, CA, USA). Chromatographic separations were carried out on a Luna® 150 mm×3 mm i.d. column packed with 3 µm CN (cyano) particles (Phenomenex®), guarded by an on-line filter. Mobile phase consist of methanol:water with sodium chloride with 0.02% of formic acid (55:45). Wavelengths for pCA and for CHD are 238 and 255 nm respectively. Influence of methanol and of sodium chloride content in the eluant has been studied. Linearity of CHD is very good, from 0.5 up to 21.2 µg/l while linearity of pCA is in the range of 0.05 to 10 µg/l with correlation coefficients above 0.999. Resolution between the components is above 4, asymmetry is about 1.3 and 1.7 for pCA and CHD respectively and the run time is less than 5 minutes. This method has been applied to CHD solution of different medical devices. No interference has been reported, and the analysis of direct injection of solution, without any treatment is achieved in less than five minutes.In conclusion, we present a validated method for dosage of CHD and its major impurity pCA, known to be carcinogen, available into medical products or medicinal device for in-vitro diagnostic.

Comparing the Effect of Irrigation With 70% Isopropyl Alcohol, Distilled Water and Saline to Remove the Residual Sodium Hypochlorite before Irrigation with Chlorhexidine in Root Canal Therapy – An In Vitro Study

“…Basrani et al (6) found a brown colour precipitate when chlorhexidine was mixed with sodium hypochlorite in the test tube and parachloraniline (PCA) was found when this precipitate subjected to gas chromatography (14,15). This precipitate is an insoluble neutral salt formed by the acid base reaction between NaOCl and CHX.…”

Development and Validation of a Novel RP-HPLC Method for the Determination of Cetrimide and Chlorhexidine Gluconate in Antiseptic Solution

“…On the contrary, an HPLC method for the determination of CHG is described in both the European Pharmacopeia [10] and the USP [16]. Furthermore, numerous analytical methods have been described for the quantification of CHG in different dosage forms, such as oral rinses, ointments, ophthalmic and skin solutions [5,17]. As for the simultaneous determination of CE and CHG, a literature survey revealed a single HPLC method, using a C-18 column (4.6 × 150 mm) and 5µm of a mobile phase consisting of 30%/55%/15% v/v/v ACN:MeOH:Phosphate buffer [18].…”

“…Chlorhexidine (CH) is the most popular substance in the category of biguanides, and due to its low toxicity, is used extensively in various commercial formulations [4]. It is a strong base that consists of two symmetrical chlorophenyl groups connected through a hydrophobic chain (Figure 1) [5]. The molecule is available in three different salts: Acetate (CHA), hydrochloride (CHH) and gluconate (CHG), with the last one being the most stable and the most soluble in water [6].…”

Graphene‐based Disposable Electrochemical Sensor for Chlorhexidine Determination

“…As a chemical anti-plaque agent, chlorhexidine is commonly utilized in mouth rinse solutions and gel formulations [2]. There are several methods of analysis for the determination of CHX reported in the literature including spectroscopic methods [3][4][5][6][7][8], HPLC methods [9][10][11][12][13][14][15][16][17][18], electrochemical methods [19][20][21][22][23][24], and capillary electrophoresis [25]. Although HPLC is a sensitive and selective technique, it is time-consuming, not eco-friendly, and is less economical than electrochemical methods as they consume large amounts of organic solvents [17,26].…”