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Sanguinarine (SAN), as the main active component of a traditional Chinese veterinary medicine, has been widely used in the animal husbandry and breeding industry. However, the metabolites of SA are still uncertain. Therefore, this research aimed to investigate the metabolites of SA based on rats in vivo. The blood, feces, and urine of rats were collected after the oral administration of 40 mg/kg SAN. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the metabolites of SAN. The elemental composition of sanguinarine metabolites was inferred by analyzing their exact molecular weight, and the structures of the metabolites were predicted based on their fragment ions and cleavage pathways. A total of 12 metabolites were identified, including three metabolites in the plasma, four in the urine, and nine in the feces. According to the possible metabolic pathways deduced in this study, SAN was mainly metabolized through reduction, oxidation, demethylation, hydroxylation, and glucuronidation. This present research has summarized the metabolism of SAN in rats, which is helpful for further studying the metabolic mechanism of SAN in vivo and in vitro. [ABSTRACT FROM AUTHOR]

Ulcerative colitis (UC) is an inflammatory bowel disease (IBD), and its pathogenesis is related to intestinal mucosal barrier damage and gut microbiota imbalance. Protopine (PRO), an isoquinoline alkaloid, is one of the main anti-inflammatory ingredients of traditional Chinese medicine Macleaya cordata (Willd.) R. Br. This study investigated the effects of PRO on the intestinal mucosal barrier and gut microbiota in dextran sodium sulfate (DSS)-induced colitis mice. C57BL/6J mice were treated with 3% DSS in drinking water to induce acute colitis, while PRO was administered orally once daily for 7 days. The results showed that PRO administration significantly alleviated the symptoms of DSS-induced colitis in mice and inhibited the expression of inflammation-related genes. In addition, PRO restored the integrity of the intestinal barrier in colitis mice by restoring colonic mucin secretion and promoting the expression of tight junction proteins. Furthermore, PRO alleviated the DSS-induced gut microbiota dysbiosis by decreasing the abundance of Proteobacteria, Escherichia-Shigella and Enterococcus, as well as enhancing the abundance of beneficial bacteria, such as Firmicutes and Akkermansia. These findings suggested that PRO effectively alleviated DSS-induced ulcerative colitis by suppressing the expression of inflammation-related genes, maintaining the intestinal mucosal barrier and regulating the intestinal microbiota. [ABSTRACT FROM AUTHOR]

Bitter orange, Citrus aurantium L. var. amara (CAVA), is an important crop and its flowers and fruits are widely used in China as a food spice, as well as in traditional Chinese medicine, due to its health-promoting properties. The secondary metabolites that are present in plant-derived foods or medicines are, in part, responsible for the health benefits and desirable flavor profiles. Nevertheless, detailed information about the bioactive ingredients in CAVA is scarce. Therefore, this study was aimed at exploring the phytochemicals of CAVA by high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Here, a systematic screening method combined with HPLC-Q-TOF-MS was presented. This technique was used to systematically screen metabolites, primarily from the complex matrix of CAVA, and to identify these compounds by their exact masses, characteristic fragment ions, and fragmentation behaviors. A total of 295 metabolites were screened by the screening method and 89 phytochemicals were identified in the flowers, fruits, roots, leaves, and branches of CAVA. For the first time, 69 phytochemicals (flavonoids, alkaloids, terpenoids, etc.) were reported from CAVA. The results highlight the importance of CAVA as a source of secondary metabolites in the food, medicine, and nutraceutical industries. [ABSTRACT FROM AUTHOR]

Aurantii fructus (zhiqiao, ZQ) is a traditional Chinese medicine (TCM) and raw material of TCM healthcare food (TCM-HF), mainly focused on the regulation of gastrointestinal disorders and the abundant application of antioxidants. Pharmacological investigations of ZQ flavonoids have identified them as the main bioactive components in recent years, but little has been reported on the extraction processes of antioxidant flavonoids (AFs). The aim of this study was to establish an efficient ultrasonic-assisted extraction (UAE) method for the extraction of AFs from ZQ using a response surface methodology (RSM), analyze the composition of AFs, and develop a qualitative evaluation method for ZQ. Flavonoid yield and antioxidant ability were selected as the responses to optimize the extraction of AFs, and the multiple effects of independent variables were investigated. The optimized conditions for the extraction of AFs based on the Box-Behnken design (BBD) were as follows: ethanol concentration, 58%; extraction temperature, 70 °C; and extraction time, 17 min. The flavonoid yield and antioxidant activity reached 241.70 mg/g and 59.42%, respectively, which matched the predicted values. Furthermore, optimized UAE processes were first established for the efficient and fast extraction of AFs. Flavanones and polymethoxyflavonoids (PMFs) were identified as potential AFs using time-of-flight mass spectrometry. Meanwhile, the quality of ZQ was evaluated using the criteria importance through intercriteria correlation (CRITIC) method for the first time, and Yuanjiang ZQ was considered as an excellent raw material of TCM-HF. [ABSTRACT FROM AUTHOR]

Edible dock, also known as Rumex patientia L.× Rumex tianschanicus A. LOS., exhibiting the characteristics of high yield, high protein and high nutrition, is widely used in the functional food industry and Chinese herbal medicine while short of secondary metabolites research. This study used UPLC-qTOF MS with two ion polarity in conjunction with Molecular Networking from Global Natural Products Social (GNPS) platform. Then, these 16 flavonoid substances including quercetin derivatives and kaempferol derivatives, firstly were identified from leaves in edible dock and were systematically investigated by UPLC-qTOF MS/MS. MS/MS experiments found that quercetin derivatives have five common characteristic ions, including m/z 300, m/z 271, m/z 255, m/z 243, and m/z 151. In addition, kaempferol derivatives have three common characteristic ions, including m/z 284, m/z 255, and m/z 227. Subsequently, 7 representative constituents were quantified via UPLC-QQQ MS with chemometrics, and the total content of 7 major flavonoids in its dry leaves is 6.988 mg/g in the 21st day samples. The work provided abundant metabolites information of flavonoids in edible dock, and the common characteristic ions will facilitate the building blocks-based molecular network (BBMN) to predict novel framework compounds from quercetin derivatives and kaempferol derivatives, laying the basis for exploitation of edible dock in the fields of pharmaceuticals, health food, and livestock feed. [Display omitted] • GNPS with HR-MS enables rapid screening secondary metabolites in edible dock. • Flavonoids in edible dock were systematically investigated by LC-MS/MS. • 16 flavonoids identified for the first time in edible dock via Molecular Networking. • The total content of main flavonoids in its dry leaves was as high as 6.988 mg•g−1. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a LC-based identification strategy was proposed and off-line two-dimensional liquid chromatography was developed for the separation and identification of flavonoids from complex samples. The highly orthogonal separation system was composed of a CD column, an OEG column and an XTerra C18 column. The CD column was employed for the first dimensional separation and the OEG column and XTerra C18 column were used for the second dimensional separation in parallel. A mixture of six traditional Chinese medicines was used as a complex sample in testing this method. Unknown peaks were identified by matching their retention times and accurate mass molecular weights with those of references. The identification efficacy of parallel unidimensional liquid chromatography and off-line two-dimensional liquid chromatography was compared, and the latter was demonstrated to be more efficient for the identification of target compounds—especially co-eluted compounds or minor compounds from complex samples. [Copyright &y& Elsevier]

Lonicerae Japonicae Caulis, the dried stem and branch of Lonicera japonica Thunb., is a Chinese Materia Medica known as Ren Dong Teng in Chinese with long use history in the traditional Chinese medicine (TCM) prescriptions. Lonicerae Japonicae Caulis possesses heat-clearing and detoxifying functions according to the TCM theory. In recent years, a large amount of experimental and clinical studies proved good anti-inflammatory effects of some heat-clearing and detoxifying herbs. The present study aims to reveal the anti-inflammatory property and functional substances of Lonicerae Japonicae Caulis. For anti-inflammatory activity test, LPS-induced RAW 264.7 macrophages, DSS-induced SPF male C57BL/6J mice model, and LPS-induced SPF male ICR mice model were used in vitro and in vivo , respectively. The behavioral changes, organ damage, and the expression of inflammatory factors such as TNT-α and IL-6 mRNA expression were measured for activity evaluation. Lonicerae Japonicae Caulis samples were prepared by solvent extraction and subsequent column chromatography. The main components were identified and determined using UPLC-UV analysis as well as NMR interpretation after purification. To testify the contribution of main components for the anti-inflammatory activity, different samples were also prepared by compound-knockout strategy. Ethanol extract of Lonicerae Japonicae Caulis could attenuate sickness symptoms in mice such as diarrhea, less activity, and depression. It could also alleviate multiple organ damage, and significantly inhibit the expression of pro-inflammatory factors such as TNF-α, IL-1β, IL-6 and IFN-γ in mice. Furthermore, the isochlorogenic acid-rich and biflavonoid-rich fractions and isochlorogenic acids A and C, and ochnaflavone could significantly down-regulate the mRNA expression of TNF-α and IL-6 in LPS-induced RAW 264.7 macrophages. Lonicerae Japonicae Caulis possesses anti-inflammatory property. Its isochlorogenic acid-rich and biflavonoid-rich fractions do the major contribution. And their main components, isochlorogenic acids A and C, and ochnaflavone, take main responsibility for the anti-inflammatory property. [ABSTRACT FROM AUTHOR]