Method Development and validation for determination of Parthenolide in mice plasma using reverse phase high performance liquid chromatography

For accurate, fast, and selective determination of PTL, a RP-HPLC method was developed and validated. The objective of the study is to develop a reverse-phase high-performance liquid chromatography (RP-HPLC) method and validate it to determine Parthenolide (PTL) in the mice plasma. Chromatographic separation was carried out using a Persil BDS (base deactivated silica) C18 column (250 mm 4.6 mm, 5 µm particle size) in isocratic mode. Flow rate was adjusted at 1.5 mL/min. The composition of the mobile phase was acetonitrile and water (55:45 v/v). Ultraviolet detection was carried out at a wavelength of 210 nm. RP-HPLC method for PTL analysis in mice's plasma and mobile phase was developed. The validation was done according to the guidelines set by ICH standards. A well-constructed chromatographic peak of PTL was obtained at 6 min, regarded as its retention time and a tailing factor of 0.998. A liquid-liquid extraction technique was used, and the PTL was extracted from plasma. Within the 5–80 µg/mL range, a high level of linearity (R2 = 0.999) was observed. Precision (Intra-day and inter-day) was conducted, and the results were not more than 2 %, indicating the method’s reproducibility was good. The plasma PTL exhibited a limit of detection (LOD) and a quantification (LOQ) of 1.596 and 5.174 µg/mL, respectively. This successful approach was utilized to ascertain PTL levels in mice plasma following validation. This method was found to be very sensitive, precise and accurate for the determination of PTL in pharmaceutical dosage form.