Quick identification of xanthine oxidase inhibitor and antioxidant from Erycibe obtusifolia by a drug discovery platform composed of multiple mass spectrometric platforms and thin-layer chromatography bioautography

Abstract

As a final step of the purine metabolism process, xanthine oxidase catalyzes the oxidation of hypoxanthine and xanthine into uric acid. Our research has demonstrated that Erycibe obtusifolia has xanthine oxidase inhibitory properties. The purpose of this paper is to describe a new strategy based on a combination of multiple mass spectrometric platforms and thin-layer chromatography bioautography for effectively screening the xanthine oxidase inhibitory and antioxidant properties of E. obtusifolia. This strategy was accomplished through the following steps. (i) Separate the extract of E. obtusifolia into fractions by an autopurification system controlled by liquid chromatography with mass spectrometry. (ii) Determine the active fractions of E. obtusifolia by thin-layer chromatography bioautography. (iii) Identify the structure of the main active compounds with the information provided by direct analysis in real time mass spectrometry. (iv) Calculate the IC50 value of each compound against xanthine oxidase using high-performance liquid chromatography. Using the caulis of E. obtusifolia as the experimental material, seven target peaks were screened out as xanthine oxidase inhibitors or antioxidants. Our screening strategy allows for rapid analysis of small molecules with almost no sample preparation and can be completed within a week, making it a useful assay to identify unstable compounds and provide the empirical foundation for E. obtusifolia as a natural remedy for gout and oxidative-stress-related diseases.

Different fingerprinting strategies to differentiate Porana sinensis and plants of Erycibe by high‐performance liquid chromatography with diode array detection, ultra high performance liquid chromatography with tandem quadrupole mass spectrometry, and chemometrics

“…As shown in Supporting Information Fig. S2, eight compounds in the fingerprints were identified by comparing with the reference compounds according to our previous work . The eight compounds were identified as 5‐caffeoylquinic acid ( 1 ), scopolin ( 2 ), chlorogenic acid ( 3 ), 4‐caffeoylquinic acid ( 4 ), scopoletin ( 5 ), 3,5‐dicaffeoylquinic acid ( 6 ), 3,4‐dicaffeoylquinic acid ( 7 ), and 4,5‐dicaffeoylquinic acid ( 8 ).…”

Chemical fingerprint of Ganmaoling granule by double-wavelength ultra high performance liquid chromatography and ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry

“…PCA and OPLS‐DA were used to describe variations in the data with minimum latent variables that were demonstrated with SIMCA‐P + (CA,USA). This facilitated grouping or classification of the fingerprints and detected outliers .…”

Analysis of pharmaceutical products and herbal medicines using ambient mass spectrometry

“…For example, CQAs in Erycibe obtusifolia [83], salvianolic acids in Danshen injection [87], and gallic acid and ellagic acid in the extract of Bergenia crassifolia L. leaves [91] were all unambiguously detected by DART-MS. Gallic acid in Liuwei Dihuang Teapill was directly analyzed by DAPCI-MS without sample pretreatment [98].…”

“…Many commercially available drugs [80] and forensic drugs [81] [83] were successfully analyzed by DART-MS using this strategy. For easily and automated manipulation, the loaded melting point tubes/Dip-it tubes were generally placed on a Dip-it sampler for introduction of samples, and the DART-MS analysis of methanol extract of Radix Aconiti Preparata [84], the boiled water extracts of Chinese star anise and toxic Japanese star anise [85], 70% aqueous ethanol of Morus alba L. [86], Danshen injection [87,88], and a herbal preparation of Sophora flavescens and Heterosmilax japonica Kunth [89] were implemented via this strategy.…”

“…For instance, DESI-MS was performed to analyze isomeranzin in Citrus aurantium L. [64]; and DART-MS was applied for analysis of various coumarin compounds in herbal medicines of Angelica tenuissima, Angelica gigas, Angelica dahurica, Cnidium officinale [78], Angelica gigas roots [79], Erycibe obtusifolia [83], and Angelicae gigantis radix [93].…”