[Analysis and evaluation on Paeoniae Radix Alba from different cultivars by UPLC-Q-TOF-MS and HPLC].

In this study, an established ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method was combined with multivariate statistical analysis to investigate the commonality and difference of main chemical components in the medicinal parts of Paeonia lactiflora from different cultivars; in addition, a high performance liquid chromatography(HPLC) method was established to simultaneously determine the content of eight active components in Paeoniae Radix Alba. Non-targeted analysis was carried out by UPLC-Q-TOF-MS on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 µm) column with a gradient elution of 0.1% aqueous formic acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 0.2 mL·min~(-1). The column temperature was 30 ℃, and an electrospray ionization source was used to acquire mass spectrometry data in positive and negative ion modes. According to the accurate molecular weight and fragment ion information provided by multi-stage mass spectrometry and by comparison with reference substances and literature reports, thirty-six identical components were identified in Paeoniae Radix Alba from different cultivars with positive and negative ion modes. In the negative ion mode, two groups of samples were well separated; specifically, seventeen components with significant differences in content were screened and identified, and one component unique in "Bobaishao" was obtained. Quantitative analysis was conducted by high-performance liquid chromatography(HPLC) on an Agilent HC-C_(18)(4.6 mm×250 mm, 5 µm) column with a gradient elution of 0.1% aqueous phosphoric acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the detection wavelength was at 230 nm. An HPLC method was developed for the simultaneous determination of eight active components(gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, galloylpaeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, benzoyl-paeoniflorin) in Paeoniae Radix Albaa from different cultivars. Satisfactory linearity was achieved within the investigated linear ranges and with fine coefficients(r>0.999 0), and the methodological investigation showed that the method had good precision, repeatability and stability. The mean recoveries were 90.61% to 101.7% with RSD of 0.12% to 3.6%(n=6). UPLC-Q-OF-MS provided a rapid and efficient qualitative analytical method for the identification of the chemical components in Paeoniae Radix Alba, and the developed HPLC method was simple, rapid and accurate, which could provide a scientific basis for the evaluation of the germplasm resources and herbal quality of Paeoniae Radix Alba from different cultivars.

["Attenuation-preservation effects" of different processed products of Aconiti Kusnezoffii Radix in industrialized production].

To study the correlation between toxicity and efficacy of different processed Aconiti Kusnezoffii Radix productsin industrial production, in order to define the optimal processing method for "attenuation-preservation effects". The HPLC method was used to determine the content of six aconitine alkaloids in Aconiti Kusnezoffii Radix and its different processed products. The Bliss method was used to determine the half-lethal dose(LD_(50)) or the maximum dose of Aconiti Kusnezoffii Radix and its different processed products in mice. The toluene-induced ear swelling method and the acetic acid twist method were used to investigate the anti-inflammatory and analgesic effects of different processed products. The results showed that: ① the total amount of diester alkaloids incrude Aconiti Kusnezoffii Radix was 0.358 8%; the total amount of diester alkaloids in Aconiti Kusnezoffii Radix prepared by pharmacopoeia-based boiling method was 0.002 2%, and the total amount of monoester alkaloids was 0.036 2%; the total amount of diester alkaloids in Aconiti Kusnezoffii Radix produced by atmospheric steaming method was 0.006 0%, and the total amount of monoester alkaloids was 0.056 7%; ② the LD_(50) of Aconiti Kusnezoffii Radix was 5.4 g·kg~(-1),and the maximum dose of processed products by two methods were 133.34 g·kg~(-1); pathological observation showed that compared with the normal group, the two kinds of processed products of Aconiti Kusnezoffii Radix had certain damage to the heart, liver and kidney; ③products processed by pharmacopoeia-based boiling method and atmospheric steaming method had anti-inflammatory and analgesic effects(P<0.01 or P<0.05). The anti-inflammatory and analgesic effects were as follows: the atmospheric steaming method was superior to the pharmacopoeia-based boiling method. The above results suggest that the crude Aconiti Kusnezoffii Radixis more toxic. The atmospheric steaming method can significantly reduce the toxicity, while retaining its good anti-inflammatory and analgesic effects, which is significantly better than the pharmacopoeia-based boiling method. The atmospheric steaming process is simple and easy to operate, and suitable for industrial production.

Polygonatum cyrtonema Hua Polysaccharide Promotes GLP-1 Secretion from Enteroendocrine L-Cells through Sweet Taste Receptor-Mediated cAMP Signaling.

Glucagon-like peptide-1 (GLP-1) secreted from enteroendocrine L-cells is a pleiotropic hormone with beneficial potential related to islet function, diet control, glucose homeostasis, inflammation relief, and cardiovascular protection. The present study aimed at investigating the effect of Polygonatum cyrtonema polysaccharide (PCP) after structural identification on GLP-1 secretion and the possible mechanism involved in the PCP-stimulated secretion of GLP-1. It was found that GLP-1 secretion was effectively promoted (p < 0.01) by PCP both in rats with oral administration for 5 weeks (13.9 ± 0.3-35.8 ± 0.3 pmol/L) and ileal administration within 2 h (13.6 ± 0.4-34.1 ± 1.1 pmol/L) and in enteroendocrine NCI-H716 cells with direct stimulation within 24 h (2.05 ± 0.3-20.7 ± 0.2 pmol/L). The sweet taste receptor T1R2/T1R3 was identified to be essential for NCI-H716 cells to directly recognize PCP. The intervention experiments showed that PCP-stimulated GLP-1 secretion was significantly depressed (p < 0.01) not only by antibodies, siRNA, and the inhibitor of T1R2/T1R3 but also by an adenylate cyclase inhibitor. These results suggest that PCP stimulates GLP-1 secretion from enteroendocrine cells possibly through activation of the T1R2/T1R3-mediated cAMP signaling pathway.

[Improvement researches on quality standard of Anemarrhenae Rhizoma and its raw processed products].

This study is to improve the quality standard and supply the scientific basis for Anemarrhenae Rhizoma and its raw processed products. Steroidal saponin including timosaponin BⅡ, timosaponin AⅢ and flavonoids including neomangiferin and mangiferin were selected as the indicative components. Silica gel G thin layer chromatography(TLC) and polyamide TLC were used to detect the two types of compounds, respectively. The contents of timosaponin BⅡ and timosaponin AⅢ were determined by HPLC-ELSD and the content of neomangiferin, mangiferin and isomangiferin were determined by HPLC-UV. Moisture, total ash and acid insoluble ash were determined according to Chinese Pharmacopoeia(2015 edition). And 80% ethanol was selected as the solvent and the content determination of total extract were determined. The fingerprints of Anemarrhenae Rhizoma and its raw processed products were established by HPLC-UV and HPLC-ELSD. The results showed that the methods of TLC and HPLC have been successfully stablished. There are 2 and 3 peaks which have been identified by HPLC-ELSD and HPLC-UV, respectively. The HPLC fingerprint methods are specific and can be used to identify and quality control for Anemarrhenae Rhizoma and its raw processed products in the mass. Comparing to Chinese Pharmacopoeia(2015 edition), the TLC identification and content determination were revised and the total extract determination and HPLC fingerprints were added in the present study. Our results can be used as the scientific basis of quqlity control for Anemarrhenae Rhizoma and its raw processed products.

[Research on hemostatic effect and mechanism of carbonized Scutellariae Radix on rats of uterine bleeding].

The aim of this paper was to investigate the hemostatic effect and mechanism of carbonized Scutellariae Radix on uterine bleeding in the rats caused by early pregnancy termination. Eight unpregnant female rats were selected as normal group. Forty female rats conceived on the same day received mifepristone(11. 4 mg·kg-1) and misoprostol(125 µg·kg-1) to induce model of incomplete abortion in early pregnancy. Abortion models were randomly divided into model group,carbonized Scutellariae Radix water extract low dose group(0. 55 g·kg-1),medium group(1. 10 g·kg-1),high dose group(2. 20 g·kg-1) and positive control group(0. 45 g·kg-1).The uterine bleeding volume was detected by ultraviolet spectrophotometry. The pathological changes of endometrium were detected by HE(hematixylin-eosin) staining. The levels of interleukin(IL-1ß),IL-6 and tumor necrosis factor(TNF-α) in the plasma of rats were determined by ELISA. The expression levels of IL-1ß,IL-6 and TNF-α mRNA in the uterus of rats were determined by RT-PCR.The protein expression levels of VEGF,MMP-2 and MMP-9 were determined by Western blot. As compared with the normal group,the uterine bleeding volume and histopathological score were increased significantly; microvessel density of endometrial tissues was decreased significantly; the contents of TNF-α,IL-1ß and the levels of TNF-α mRNA and IL-1ß mRNA in the plasma were increased,while the content of IL-6 and level of IL-6 mRNA were decreased significantly. The protein expression levels of VEGF,MMP-2 and MMP-9 in the uterine tissues were also decreased. As compared with the model group,the uterine bleeding volume was decreased significantly in the carbonized Scutellariae Radix medium dose and high dose groups; endometrial repair was promoted,and the microvessel density of endometrial tissues was increased significantly; the contents of IL-1ß and TNF-α in the plasma of rats were decreased significantly,while the content of IL-6 in the plasma of rats was increased significantly; the expression levels of IL-1ß and TNF-α mRNA in the uterus of rats were decreased and the expression level of IL-6 mRNA showed an increase; the protein expressions of VEGF,MMP-2 and MMP-9 were decreased significantly in carbonized Scutellariae Radix medium and high dose groups. In conclusion,carbonized Scutellariae Radix showed good hemostatic effect,and its mechanism may be related to the repair of endometrium and inhibition of inflammatory reaction.

Alkaloids from flowers of Erythrina corallodendron.

Two new Erythrina alkaloids, 10-oxo-erythrinine (1) erythrinine N-oxide (2) together with 23 known ones were obtained from the flowers of Erythrina corallodendron. The structures were determined based on analysis of their spectroscopic data. All compounds were first isolated from plants of Erythrina corallodendron.

[The influence of different processing on the content of flavones of Dendranthema morifolium].

OBJECTIVE: To investigate the effect of different processing methods on the content of luteolin-7-O-beta-D-glucoside, acacetin-7-O-beta-D-glucoside, luteolin, and acacetin of Dendranthema morifolium. METHODS: The optimal conditions were achieved on phenomenex, synergi Fusion-RP (5 microm, 4.6 mm x 250 mm)analytical column with a gradient mobile phase consisting of methanol and 0.1% phosphoric acid and detection wavelength set at 350 nm. RESULTS: The significant effect of different processing methods on the content of luteolin-7-O-beta-D-glucoside, acacetin-7-O-beta-D-glucoside, luteolin and acacetin was proved. CONCLUSION: Different processing methods have significant effect on flavonoids of Dendranthema morifolium, it should be paid attention to drying temperature while sulphur fumigation.

[Chemical constituents of Carya cathayensis and their antitumor bioactivity].

OBJECTIVE: To investigate the chemical constituents of Carya cathayensis and their antitumor bioactivity. METHODS: The compounds were isolated by Sephadex LH-20 and silica gel column chromatography. Their structures were identified by physicochemical properties and spectroscopic analysis. Then their cytotoxic activity was studied. RESULTS: Five compounds were elucidated as chrysophanol (1), physcion (2), beta-sitosterol (3), pinostrobin(4), 4,8-dihydroxy-1-tetralone (5). CONCLUSION: Compounds 2 and 5 are isolated from Carya cathayensis for the first time. In the MTT antitumor experiments, the compounds 1,4 and 5 have the cytotoxic activity to KB cell.

[Study on industrialization production technology of decoction pieces shuangfei of Platycodon grandiflorum].

OBJECTIVE: To establish industrialization production technology parameter for decoction pieces shuangfei of Platycodon grandiflorum. METHODS: Taking platycodin D, polysaccharide and the appearance of the decoction pieces shuangfei of Platycodon grandiflorum as the quality evaluation indexes, we investigated the size of Platycodon grandiflorum, the best slicing extent, the best slicing thickness, drying temperature and drying time. RESULTS: The optimal processing was as follows: the diameter of Platycodon grandiflorum was about 1.5 cm, moistend the selected Platycodon grandiflorum until the water content was 55% - 60%. Then rolled them into 1.5 cm thickness pieces. Dryed them at 60 degrees C for 8 - 9 h and packed them after they were cold. CONCLUSION: The industrialization production technology for decoction pieces shuangfei of Platycodon grandiflorum is stable and the efficiency is improved.

[Study on the chemical composition of triterpenoid from the fruit of Buddleja lindleyana and their neuroprotective activitiy].

OBJECTIVE: To study the chemical composition of triterpenoid from the fruit of Buddleja lindleyana. METHODS: The chemical components were isolated by chromatography. The structures were identified by spectral data. The neuroprotective activity of these compounds were evaluated by using MPP+ induced injury in PC12 cells. RESULTS: 3 compounds were separated and identified as oleanane, alpha-L-msnnopyranoside derive (1), 13, 28-epoxy-3beta,23-dihydroxy-11-oleanene (2), 3, 23, 28-trihydroxyolean-11,13 (18)-diene (3). Compounds 1-3 showed obviously neuroprotective activity. CONCLUSION: The data of compound (1) is reported for the first time. The neuroprotective activities of compounds 1, 2, 3 are reported for the first time.