Chemical profiling and quantitative analysis on the aqueous extract of Pimpinella anisum fruit by liquid chromatography-tandem mass spectrometry.

Anise fruit (Aniseed) has been used for many years as a traditional medicine in various countries throughout the world; however, the chemical material basis of Aniseed water extract (AWE) has not been examined in detail, limiting our understanding of its pharmacological mechanism and methods for practical quality control. A high-efficiency and high-sensitivity LC-triple time-of-flight tandem mass spectrometry (MS/MS) analysis method using data processing method combined with product ion and neutral loss filtering for systematic screening and identification of the constituents of AWE was established. A quantitative method was established by using LC-MS/MS with multiple reaction monitoring for 10 min to determine the concentration of 17 representative constituents. A total of 89 compounds were discovered in AWE, of which 31 were confirmed by the reference standards, while 24 were found in Aniseed for the first time. The qualification analysis results showed that chlorogenic acids and luteolin derivatives were the major compounds. The linearity, sensitivity, precision, stability, repeatability, and accuracy of the method were verified, which demonstrated that the method could meet the requirements for quantification. This work contributes to a better understanding of the chemical material basis of Aniseed and assists in the development of effective analytical methods for natural medicines.

Comprehensive chemical profiling and quantitative analysis of ethnicYi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS.

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

Exploring the effect of the Uyghur medicine Munziq Balgam on a collagen-induced arthritis rat model by UPLC-MS/MS-based metabolomics approach.

ETHNOPHARMACOLOGICAL RELEVANCE: Munziq Balgam (MBm) is a classic preparation of a traditional Uyghur medicine used for many years to treat abnormal body fluid diseases. The formula, as an in-hospital preparation, has already been used in the Hospital of Xinjiang Traditional Uyghur Medicine to treat rheumatoid arthritis (RA) with significant clinical effects. AIM OF THE STUDY: This study intends to reveal the intervention effect of MBm on collagen-induced arthritis (CIA) rats, discover the potential biomarkers with efficacy, and explore the mechanisms of metabolic regulation by using metabolomics method. MATERIAL AND METHODS: Sprague Dawley (SD) rats were randomly divided into five groups: blank group, CIA model group, Munziq Balgam nomal-dosage, Munziq Balgam high-dosage group and control group. Body weight, paw swelling, arthritis index, immune indices and histopathological experiments were carried out. Plasma from rats were detected by UPLC-MS/MS. Metabolomics of plasma was performed to analyze metabolic profiles, potential biomarkers, and metabolic pathways of MBm for CIA rats. The main metabolic result of Uyghur medicine MBm was compared with that of Zhuang medicine Longzuantongbi granules (LZTBG) to explore the characteristics of two ethnic medicines from different regions for RA. RESULTS: MBm could significantly alleviate symptoms of CIA rats by relieving arthritis symptoms on paw redness and swelling, inflammatory cell infiltration, synovial hyperplasia, pannus, cartilage and bone tissue destruction, as well as inhibiting the expression of IL-1ß, IL-6, TNF-α, UA and ALP. Linoleic acid, alpha-linolenic acid, pantothenate and CoA biosynthesis, achidonic acid, gycerophospholipid, sphingolipid metabolism, primary bile acid biosynthesis, porphyrin and chlorophyll metabolism and fatty acid degradation served as the main nine pathways of the interventional effect of MBm on CIA rats. Twenty-three different metabolites were screened out and strongly associated with the indicator makes of RA. Eight potential efficacy-related biomarkers were finally discovered in metabolic pathway network (phosphatidylcholine, bilirubin, sphinganine 1-phosphate, phytosphingosine, SM (d18:1/16:0), pantothenic acid, l-palmitoylcarnitine, chenodeoxycholate). Three metabolites (chenodeoxycholate, hyodeoxycholic acid and O-palmitoleoylcarnitine) were changed in both the metabolic study of MBm and LZTBG intervention effects on CIA rats. Additionally, MBm and LZTBG shared the same 6 metabolic pathways including linoleic acid, alpha-linolenic acid, pantothenate and CoA biosynthesis, achidonic acid, gycerophospholipid, and primary bile acid biosynthesis. CONCLUSION: The study suggested that MBm may effectively alleviate RA by regulating inflammation, immunity-related pathways and multiple targets. Metabolomics analysis showed that MBm (Xinjiang, the north of China) and LZTBG (Guangxi, the south of China), two ethnic medicines from different regions in China, share common metabolites and pathways but also have distinct differences in their interventions for RA.

Metabolomics reveals the intervention effect of Zhuang medicine Longzuantongbi granules on a collagen-induced arthritis rat model by using UPLC-MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is known as "Fawang" in Zhuang medical theory. Longzuantongbi granules (LZTBG) is an in-hospital preparation used at the First Affiliated Hospital of the Guangxi University of Chinese Medicine. This medicine is based on traditional Zhuang medicine theory for the treatment of "Fawang", and has an effectiveness of over 86.67%. It comprises eight medicinal materials, including the main drug Toddalia asiatica (L.) Lam. and Kadsura coccinea (Lem.) A.C. Smith, the assisting drugs Alangium chinense (Lour.) Harms, Zanthoxylum nitidum (Roxb.) DC., Sinomenium acutum (Thunb.) Rehd.et Wils., Bauhinia championii (Benth.) Benth., Spatholobus suberectus Dunn, and Ficus hirta Vahl. All of these herbs are commonly used in Zhuang medicine. AIM OF THE STUDY: This study aims to reveal the effect of LZTBG on collagen-induced arthritis (CIA) rats, to discover the potential efficacy-related biomarkers and explore the intervention mechanism of LZTBG from a molecular level, based on metabolomics. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were randomly assigned into a normal group, a CIA model group, a positive control (MTX) group and two different LZTBG treatment groups (5.4 g/kg/d and 2.7 g/kg/d). Body weight, arthritis index (AI), paw swelling, and hematoxylin and eosin (HE) staining experiments were used to evaluate the efficacy of the established model. A metabolomics method based on an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique was established to analyze plasma taken from the rats, and to explore the interventional mechanism of LZTBG. RESULTS: LZTBG showed a positive effect on the CIA model rats. Thirty-one differential metabolites were screened out, and combined with pathway analysis, 11 potential efficacy-related biomarkers were then mapped in the pathway. These included linoleic acid (LA), phosphatidylcholine (PC), lysophosphatidylcholine (LPC), arachidonic acid (AA), 12-HETE, alpha-linolenic acid (ALA), 13(S)-HOT, 2-oxobutanoate, 3-hydroxybutyric acid, L-Valine, and acetylcholine. Furthermore, it was found that these metabolites may exhibit an intervention effect by means of modulating pathways related to both lipid metabolism and amino acid metabolism to associated with inflammation. CONCLUSION: LZTBG can effectively alleviate symptoms of RA, an effect that can primarily be attributed to the regulation of multiple pathways and multiple targets These results demonstrate that LC-MS/MS-based metabolomics is an advantageous technique for the investigation of the intervention effect and molecular mechanism of traditional compound medicine.