Construction and Chemical Profile on "Activity Fingerprint" of Citri Reticulatae Pericarpium from Different Cultivars Based on HPLC-UV, LC/MS-IT-TOF, and Principal Component Analysis.

Citri Reticulatae Pericarpium (CRP), known as Chenpi (CP) in Chinese, is a medicinal food for health and fitness. In order to find out the characteristic activity chemicals distinguishing various cultivars of CRP and provide a reference for effective development of citrus resources, an "activity fingerprint" of CRP from 21 different cultivars was established based on the evaluation of antitussive and expectorant activities. There were 18 common peaks in the HPLC fingerprint, of which 3 flavonoid glycosides and 14 polymethoxyflavonoids (PMFs) were identified by LC/MS-IT-TOF. Furthermore, five characteristic chemicals were determined and similarity calculation with principal component analysis (PCA) was combined together to compare the similarity and difference among various cultivars. The results showed that some other cultivars were also similar to C. reticulata "Chachi" except for C. reticulata "Tangerina" and C. reticulata "Dahongpao" recorded in Chinese Pharmacopoeia. Most importantly, the peels of C. reticulata "Shiyueju," C. reticulata "Ponkan," C. reticulata "Tribute," and C. reticulata "Bayueju," traditionally rarely used for medicinal food, were highly similar to that of C. reticulata "Chachi" and rich in bioactive flavonoids, which can be considered the effective medicinal resources of CRP.

Identification and determination of chemical constituents of Citrus reticulata semen through ultra high performance liquid chromatography combined with Q Exactive Orbitrap tandem mass spectrometry.

Citrus reticulata semen, a traditional Chinese medicinal material, has desirable medicinal and dietary properties. In this study, a method combining ultra high performance liquid chromatography with Q Exactive Orbitrap tandem mass spectrometry was established and validated for the identification and analysis of the chemical components of C. reticulata semen for the first time. The evaluation of different retention times and fragmentation characteristics, as well as comparative analysis with the literature, resulted in the identification of 35 chemical constituents, including 21 flavonoids and 14 other compounds. The 21 flavonoids derived from C. reticulata semen were reported for the first time. Seven of the chemical components of C. reticulata semen were quantitatively analyzed using the developed method under the optimal conditions. The results showed that the content of limonin, hesperidin, nobiletin, synephrine, tangeretin, 3,5,6,7,8,3',4'-heptamethoxyflavone and 5-hydroxide-6,7,8,3',4'-pentamethoxyflavone in C. reticulata semen was 11.1666, 0.0404, 0.0092, 0.0255, 0.0087, 0.0010, and 0.0008 mg/g, respectively. This study demonstrated that the ultra high performance liquid chromatography Q Exactive Orbitrap mass spectrometry based method can be used to rapidly and reliably analyze the chemical constituents of C. reticulata semen. These results provide a scientific basis for further studies of C. reticulata semen.

Andrographolide inhibits proliferation and induces apoptosis of nasopharyngeal carcinoma cell line C666-1 through LKB1-AMPK-dependent signaling pathways.

BACKGROUND: Andrographolide (Andro) belongs to the main bioactive ingredients of Andrographis paniculata. Many studies have shown that andro has a variety of pharmacological activities such as anti-inflammatory, anti-bacterial, anti-virus, anti-oxidant, immune regulation and liver protective effects. Moreover andro has been reported to have anticancer activity in multiple types of cancer, including gastric cancer, breast cancer, lung cancer and so on. However, there is no report about the effect of andro on the human NPC cell line C666-1 and the molecular mechanisms of andro-mediated apoptosis in C666-1 cells remain to be clarified. METHODS: Cell proliferation was measured by a CCK8 assay, cell apoptosis rate was evaluated by flow cytometric analysis, and the protein expression of LKB1/AMPK signaling pathways was detected by Western blotting. RESULTS: Treatment with andro inhibited cell proliferation and induced apoptosis of C666-1 cells. Moreover, andro could activate LKB1-AMPK signaling. We also demonstrated that Ca2+/calmodulin-dependent protein kinase kinaseß (CaMKKß) was not involved in the regulation of andro on AMPK activation in C666-1 Cells. CONCLUSIONS: Andro suppressed proliferation and induced apoptosis of C666-1 cells through regulating the LKB1/AMPK/mTOR signal pathway.

Evaluation of chemical components in Citri Reticulatae Pericarpium of different cultivars collected from different regions by GC-MS and HPLC.

To discriminate the feasible differences and find potential similarities and relationships of Citri Reticulatae Pericarpium (CRP), this work was accomplished by a comprehensive and reliable method using gas chromatography-mass spectrometer (GC-MS) to analyze the volatile oils and high-performance liquid chromatography (HPLC) simultaneously to determine the contents of five bioactive flavonoids, namely hesperidin, nobiletin, 3,5,6,7,8,3',4'-heptamethoxyflavone, tangeretin, and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone in 25 batches of CRP samples of 10 cultivars collected from different regions in China. The GC-MS analyses indicated that 98 compounds were successfully identified from the volatile oils obtained and the major constituents of volatile oil are d-limonene, γ-terpinene, α-pinene, linalool, and myrcene. Even 2-(methylamino) benzoate was found in all cultivar samples harvested at maturation stage. Under the optimal condition, the quantitative analyses of five bioactive flavonoids were successfully performed by HPLC and hierarchical cluster analysis (HCA). Results showed significant differences among cultivars in the contents of five bioactive flavonoids mentioned earlier. The HCA and GC-MS results provided a convenient approach which might be applied for rapid similarity evaluation and also holds the potential for analysis of compounds present in other plants. Therefore, this work obtained offers scientific basis to control quality and develop medicinal value of the medicinal materials in CRP.