Determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations by liquid chromatography with mass spectrometry

Abstract

So far, the components responsible for the neuroprotective effects of Calculus bovis are unclear. Cholesterol, one of the major components in Calculus bovis, is easily oxidized into oxysterols, which possess direct or indirect neuroprotective effects proved by our and others' previous studies. Therefore, a liquid chromatography with mass spectrometry method coupled with ultrasonic extraction and solid-phase extraction was developed for the determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations. Chromatographic separation was achieved on a C18 column with isocratic elution at a flow rate of 1 mL/min. The established method showed good linearity (R(2) > 0.998), sensitivity with low limits of detection (0.06-0.39 μg/g), acceptable precisions (relative standard deviations ≤ 7.4%), stability (relative standard deviations ≤ 5.9%), and satisfactory accuracy (92.4-102.9%) for all analytes identified by different retention times, which could be applied for the determination of oxysterols. Five kinds of oxysterols proved to function as neuroprotectants were detected at different concentrations. Among them, 7β-hydroxycholesterol and cholestane-3β,5α,6β-triol were rather abundant in the samples. It could be concluded that the potential neuroprotective components in Calculus bovis may be these oxysterols.

Shexiang Baoxin Pill, a Formulated Chinese Herbal Mixture, Induces Neuronal Differentiation of PC12 Cells: A Signaling Triggered by Activation of Protein Kinase A

“…This drug-pair prevented neurons from apoptosis and relieved the brain damage. Bovis Calculus Artifactus has been used for more than 2,000 years in TCM practice, and one of the major components is the oxygenated derivative of cholesterol-oxysterol, a chemical being shown to possess neuroprotective effects (Wang et al, 2015). Cinnamomi Cortex showed brain protective function by increasing the activity of superoxide dismutase and decreasing the oxidative stress (Lv, 2015).…”

Quantification of sterols from carp cell lines by using HPLC–MS

“…In contrast, the latter consist of less preparation steps resulting in faster analysis and a lower probability of artifact generation [23]. HPLC separations of the target analytes from various matrixes, using normal as well as RP chromatography combined with ultra violet (UV) [24][25][26][27], refractive index (RI) [28] or MS detection [29][30][31][32][33][34][35][36] have been previously reported. Application of MS detection not only allows the detection of all COPs, including those that completely lack chromophores like 5,6-epoxy-cholesterol, but is also more sensitive in comparison to the optical detection methods, reaching detection limits for some COPs as low as 10 ng/mL [31].…”

“…Although several methods for cholesterol and oxysterol analysis have already been published [29][30][31][32][33][34][35][36], no application for fish cell lines in culture has been reported yet. Hence, we report a fast and sensitive quantification method for cholesterol and oxysterols in fish cell cultures consisting of RP-HPLC separation coupled to APCI-MS detection.…”

Efficient discovery and capture of new neuronal nitric oxide synthase–postsynaptic density protein‐95 uncouplers from herbal medicines using magnetic molecularly imprinted polymers as artificial antibodies

“…The literature and clinical evidence suggests that the Chinese herbal medicines commonly used in stroke therapy are both neuroprotective and have few to no adverse effects . Herbal medicines have been used to prevent and treat diseases for thousands of years because of their high efficacy, low toxicity, and low costs . However, herbal medicines are complex mixtures with many components , making the extraction of active compounds very time‐consuming and labor‐intensive isolation.…”