Application of a Validated RP-HPLC Method in Solubility and Dissolution Testing for Simultaneous Estimation of Diacerein and Its Active Metabolite Rhein in Presence of Coformers in the Eutectic Tablet Formulation

Abstract

The present study aimed to develop and validate a novel reversed-phase high-performance liquid chromatography method for simultaneous estimation of Diacerein (DIA) and Rhein (Rh, alkaline degradation product and active metabolite) in the presence of various coformers used to prepare eutectic oral formulation. Chromatographic separations were achieved on a Phenomenex Gemini C18 column (250 mm × 4.6 mm, 5 μm) placed in the thermostated column oven at 40°C. The mobile phase, comprising of acetonitrile and 10 mM ammonium acetate (pH 3.0), was eluted through the gradient system with 0.8 mL/min flow rate at 254 nm detection and analytical run time of 14 min. Additionally, the method was validated for specificity, linearity, precision, accuracy, selectivity, limit of quantitation, limit of detection and robustness as per International Conference on Harmonization guideline. The developed method was applied for the comparison of drug release profiles of pure DIA and from prepared eutectic formulations for the quantitation of DIA and Rh in the multicomponent adducts. The achieved method advocated their applicability in routine quality control analysis of DIA formulations without interference of degraded product and excipients.

RP‐HPLC method development, validation and pharmacokinetic applicability in preclinical evaluation of rhein treated with novel diacerein eutectics

“…The eutectics of DIA with DA and FA were formulated as described previously (Patel, Raval, & Pethani, 2020; Patel, Raval, & Sheth, 2020). The prepared eutectics were characterized and analyzed using differential scanning calorimetry (DSC), powder X‐ray diffraction (PXRD), Fourier transmission infrared spectrophotometry (FT‐IR), and scanning electron microscopy (SEM) as described previously (Patel & Raval, 2022; Patel, Raval, & Pethani, 2020; Patel, Raval, Pethani, & Sheth, 2020; Patel, Raval, & Sheth, 2020).…”

“…Before the experiment, the animals were acclimatized for a week and then fasted overnight with free access to water. The experimental protocol was approved by the Institutional Animal Ethics Committee on 12 December 2016 (IAEC/DPS/SU/1609; Patel, Raval, & Pethani, 2020). Rats were randomly distributed into three groups ( n = 12).…”

“…Rats were randomly distributed into three groups ( n = 12). Each group was administered an orally pure DIA suspension, DIA–DA and DIA–FA eutectic suspensions in water containing 0.2% w/v sodium carboxymethylcellulose as a suspending agent at a dose equivalent to 30 mg/kg body weight of DIA, respectively (Patel, Raval, & Pethani, 2020; Tamura et al, 2002). Following the drug treatment, blood samples were collected through the retro‐orbital plexus route using heparinized tubes at predetermined time intervals, 0 (predose), 0.25, 0.5, 0.75, 1, 1.5, 2, 4, 6, 12 and 24 h following oral administration.…”

“…Therefore, we attempted to develop and validate a simple and effective RP‐HPLC technique for estimating DIA in rat plasma. In line with this, we synthesized and characterized new DIA eutectics with fumaric acid (FA) and 2,4‐dihydroxy benzoic acid (DA) (Patel, Raval, & Pethani, 2020; Patel, Raval, & Sheth, 2020). Moreover, the proposed RP‐HPLC method was validated as per the United States Food and Drugs Administration (USFDA) guidelines and applied for quantifying Rh in rat plasma.…”

“…In this regard, reversed‐phase high‐performance liquid chromatography (RP‐HPLC) has been utilized to measure the drugs in biological matrices, which provides robust and accurate results for pharmacokinetic measurements (Prathap et al, 2013; Smolec et al, 2005). As DIA is metabolically transformed to Rh, only Rh has been quantified in biological fluids to evaluate its pharmacokinetics (Patel, Raval, & Pethani, 2020). Very few analytical methods have been reported for the analysis of Rh in biological matrices, including RP‐HPLC with various detection systems (Chakrabarty et al, 2008; Dan & Xinhui, 2003; Duan et al, 2013; Ojha et al, 2009; Tang et al, 2007; Yaroshenko et al, 2014), liquid chromatography–tandem mass spectrometry (Hou et al, 2014; Jiang et al, 2012; Layek et al, 2008; Lee et al, 2003; Lv et al, 2008; Yi et al, 2006) and RP‐HPLC with fluorescence detection (Mohammed et al, 2020; Wan et al, 2013; Zhang et al, 2005).…”