Background
A precise, simple, accurate, and quick HPLC–PDA method for the determination of eluxadoline and rifaximin in rat plasma was developed and validated in this study. In this method, Loperamide hydrochloride was used as the internal standard and plasma samples were prepared using a liquid–liquid extraction technique for which acetonitrile was a solvent. An Agilent Symmetry C8 column (5 µm, 250 mm × 4.6 mm) at 283 nm and isocratic elution using HPLC grade acetonitrile and 7 mM TEA (pH 2.5) with a ratio of (40: 60 v/v) was used as a mobile phase and the flow rate employed was 1 mL min−1. A satisfactory chromatographic separation was accomplished.
Results
An HPLC–PDA method for the determination of eluxadoline and rifaximin with retention times of 3.06 and 7.82 min, respectively, was developed. The calibration curves appear linear for both eluxadoline and rifaximin in the range of 5–200 ng mL−1 and 10–400 ng mL−1, and the corresponding correlation coefficient values were found to be 0.9999 and 0.9998 respectively. Lower limits of quantification (LLOQ) for eluxadoline and rifaximin were evaluated to be 5.0 ng mL−1 and 10.0 ng mL−1, respectively. The accuracy and precision results in all validation experiments were within the acceptance limits of FDA guidelines.
Conclusion
The developed HPLC–PDA approach was fully validated to meet the USFDA guidelines for bioanalytical method validation in terms of precision, accuracy, and stability. The presented approach could be beneficial for the determination of ELX and RFX in rat plasma, according to validation parameters. This is one of the efficient method to study the pharmacokinetics of ELX and RFX in rats.
Graphical abstract
Benzothiazole (BTZ)-zinc porphyrin (ZnP) dyads, Dyad-1 and Dyad-2 connected together with two different spacers, ester and ethoxy esters, were synthesized and light induced energy and electron transfer events were investigated. Within these dyads, due to the spectral overlap of the BTZ emission with the ZnP absorption, a selective photoexcitation of BTZ at 325 nm resulted in the photo-induced energy transfer (PEnT) from 1BTZ* to ZnP displaying the quenching of the BTZ emission followed by the concurrent appearance of the ZnP emission at 600 and 650 nm suggesting the formation of the 1ZnP* [Formula: see text]. 1BTZ*-ZnP [Formula: see text] BTZ-1ZnP*. When the dyads are titrated with imidazole appended fullero[C[Formula: see text]/C[Formula: see text]]pyrrolidines, four supramolecular triads, involving the axial co-ordination of the imidazole to the zinc center of the ZnP, were formed and the assembly formation was systematically monitored by the optical absorption technique. Cyclic voltammetry and the density functional theory calculations have revealed that, in these triads, the zinc porphyrin acts as an electron donor and fullerene moiety as the electron acceptor. Steady state fluorescence studies revealed that, upon selective excitation of the ZnP moiety at 550 nm, the emission of ZnP at 600 and 650 nm was quenched revealing the occurrence of photo-induced electron transfer (PET) from 1ZnP* to fullerene moiety leading to the formation of charge separated state [Formula: see text]. BTZ-1ZnP* : (ImC[Formula: see text] BTZ-ZnP[Formula: see text]:(ImC[Formula: see text]. More importantly, when the supramolecular triads were excited at 325 nm, the wavelength at which the BTZ absorbs predominantly, the emission of the BTZ moiety which was quenched due to PEnT from 1BTZ* to ZnP followed by the PET from 1ZnP* to fullerene indicates the probability of occurrence of 1BTZ*-ZnP:(ImC[Formula: see text] [Formula: see text] BTZ-1ZnP*[Formula: see text]: (ImC[Formula: see text] BTZ-ZnP[Formula: see text]:(ImC[Formula: see text].
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