Simultaneous HPLC determination of ergosterol and 22,23‐dihydroergosterol in Flammulina velutipes sterol‐loaded microemulsion

Abstract

An efficient HPLC method was developed and validated for the simultaneous determination of ergosterol and 22,23-dihydroergosterol in Flammulina velutipes sterol-loaded microemulsions (FVSMs). The different chromatographic conditions for in vitro and in vivo determinations were investigated, with the application examined by tissue distribution. Chromatographic separation was achieved on an Inertsil ODS-SP (250 × 4.6 mm, 5 µm) analytical column using a mobile phase of 98% methanol (in vitro), and 93% methanol for stomach samples and 96% methanol for other samples (in vivo) at 1.0 mL/min. The sterol content was detected at 282 nm. The established in vitro linearity ranges for ergosterol and 22,23-dihydroergosterol were 0.58-72.77 µg/mL (r1 = 0.9999) and 0.59-73.25 µg/mL (r2 = 0.9999), respectively, with the biological (in vivo) samples following the same trend. The accuracy of the method was >99% (in vitro) and between 93%-108% (in vivo). The LOQ was 2.15 µg/L for ergosterol and 2.41 µg/L for 22,23-dihydroergosterol in the in vitro studies. Also, the precisions met the acceptance criterion. These results indicate that the established HPLC method was specific, linear, accurate, precise and sensitive for the separation and simultaneous determination of ergosterol and 22,23-dihydroergosterol.

In depth understanding the molecular response to the enhanced secretion of fatty acids in S accharomyces cerevisiae due to one-step gene deletion of acyl-CoA synthetases

“…Ergosterol was quantified by HPLC (Agilent 1200) equipped with a 4.6 × 150 mm XDB-C18 columns as previously described [26,27]. Methanol: water (97: 3, v/v) was used as mobile phase with a flow rate of 0.8 mL min −1 .…”

Determination of serum brassicasterol in spontaneously hypertensive rats stroke‐prone fed a high‐ergosterol diet by ultra performance liquid chromatography

“…On the basis of several investigations, this method is established as the first UPLC‐UV method applicable to the simultaneous determination of serum ergosterol, brassicasterol, and cholesterol. Since LC‐APCI‐MS/MS and HPLC‐UV are the existing methods for the determination of serum ergosterol, the present method is an alternative tool to measure serum ergosterol together with brassicasterol and cholesterol, which is less expensive to LC‐MS/MS. Moreover, from the animal experiment, it is evident that a high‐ergosterol diet induces the elevation of serum brassicasterol in SHRSP rats, which are a well‐known animal model of phytosterolemia, suggesting a possibility that the absorption of ergosterol as a precursor of brassicasterol is occurred in SHRSP rats.…”

“…LC‐APCI‐MS/MS is highly sensitive and selective, resulting in a crucial tool for the quantitative lipid analysis in biological samples. The other method is by conventional HPLC‐UV, which was developed for the determination of ergosterol in microemulsoins and their tissue distribution in mice . In our previous study, high‐performance liquid chromatography with electrochemical detection (HPLC‐ECD) was suitable to the determination of serum phytosterols in rats , while ultra performance liquid chromatography with UV detection (UPLC‐UV) was applicable to the determination of cellular phytosterols in CaCo‐2 cells .…”

Stability‐indicating HPLC method for acyclovir and lidocaine in topical formulations

“…Additionally, combination formulations require a broad class of method validation steps to monitor the stability of the product over time. Further, adequate separation of the degradation products from the target analyte(s) needs optimization of several chromatographic factors, including column packing, mobile phase composition, the elution mode, and other variables (Tong et al, 2014;Veillette, Winans, Forland, & Maskiewicz, 2016;Yaseen Malik et al, 2017).…”