Improvement Effects of Alisol B 23-acetate on Glycolipid Metabolism Disorder in Obesity Model Mice / 中国药房

OBJECTIVE：To st udy the improvement effects and its mechan ism of alisol B 23-acetate on glycolipid metabolism disorder in obesity model mice. METHODS ：The mice was given high-fat diet for 10 weeks to induce obesity model. Model mice were randomly divided into model group ，orlistat group （positive control ，15.6 mg/kg）， alisol B 23-acetate low-dose ， medium-dose and high-dose groups （7.5，15，30 mg/kg），with 10 mice in each group. Another 10 mice fed with normal diet were set as normal group. The mice in normal group and model group were given water intragastrically ，and administration groups were given the corresponding drugs intragastrically ，with the volume of 20 mL/kg，once a day ，for consecutive 4 weeks. After last medication，body weight ，waist circumference ，body fat ，muscle and body fluid mass were measured ；the serum levels of blood lipids indicators （TC，TG，HDL-C，LDL-C）and blood glucose were determined. The levels of PPAR-γ，NF-κB and IL-6 in liver tissue as well as serum level of TNF-α were determined by ELISA. The pathomorphological changes of visceral fat and liver tissue in mice were observed by HE staining. RESULTS ：Compared with normal group ，body weight ，waist circumference ，body fat and body fluid mass were significantly increased in model group （P＜0.01）；serum levels of TC ，TG，HDL-C，blood glucose and TNF-α，the levels of PPAR-γ，NF-κB and IL-6 in liver tissue were increased significantly （P＜0.05 or P＜0.01）；the structure of adipocytes was ruptured ，the volume of adipocytes was increased ，accompanied by inflammatory cell infiltration ；a large number of liver cells were edema ，and cytoplasm was loose and light stained，accompanied by fatty degeneration. Compared with model group ，the body weight ，body fat and body fluid mass as well as serum le vels of TG and TNF-α in alisol B 23-acetate groups were significantly reduced （P＜0.01）；the levels of TC and blood glucose in serum ，IL-6 in liver tissue were significantly decreased in alisol B 23-acetate medium-dose and high-dose groups （P＜0.05 or P＜0.01），and the level of PPAR-γ in liver tissue was increased significantly（P＜0.05 or P＜0.01）； the waist circumference and NF-κB levels in liver tissue in alisol B 23-acetate high-dose group were decreased significantly （P＜ 0.01）；serum level of HDL-C in alisol B 23-acetate medium-dose group were decreased significantly （P＜0.01）；the adipocytes were closely arranged and small in size ；the hepatocytes were mild to moderate swelling ，a small amount of cytoplasm was loose ， light stained or vacuolated ，and a small number of hepatocytes were accompanied by steatosis and small focal infiltration of inflammatory cells. CONCLUSIONS ：Alisol B 23-acetate can improve the disorder of glucose and lipid metabolism in obesity model mice ，and its mechanism may be related to the regulation of PPAR-γ，NF-κB，IL-6 levels in liver tissue and TNF-α levels in serum.

Analysis on material basis and efficacy network of Huopo Xialing Decoction in treating damp pathogen stagnation of lung syndrome of early COVID-19 / 中草药

Objective: To screen the material basis of Huopu Xialing Decoction in the treatment of damp pathogen stagnation of lung syndrome of early COVID-19 and predict its mechanism. Methods: Literatures and clinical reports were reviewed to analyze the relationship between Huopu Xialing Decoction and damp pathogen stagnation of lung syndrome of early stage of COVID-19. TCMSP database was used to screen the potential active components in Huopo Xialing Decoction. Molecular docking of the active components with SARS-CoV-2 hydrolase and ACE2 was also carried out. According to the binding ability, the core components with both were screened. The interaction network of key components target proteins was constructed by using the software of Cytoscape to screen the main targets; The GO analysis of the main targets was carried out by using the STRING database, and the Pathway and KEGG enrichment analysis was carried out by using the plug-in of the software ClueGO of Cytoscape. Results: The Huopo Xialing Decoction was used to treat the early coronavirus pneumonia with damp pathogen and lung depression syndrome in the relationship analysis between prescriptions and syndrome, and the potential components of the 12 ingredients in Huopo Xialing Decoction were selected, with 67 core targets. Among them, paryriogenin I from Tetrapanax papyrifer, ergosterol peroxide from Poria cocos and Polyporus umbellatus, baicalin from Pinellia ternata had good binding activity with SARS-CoV-2 3CL hydrolase and ACE2. The results of enrichment analysis of GO, Pathway and KEGG showed that 12 potential components in Huopo Xialing Decoction were involved in regulating the biological processes such as stimulation response, signal transduction, cell death and the related signaling pathways including interleukins, EGFR in cancer, tyrosine kinases, programmed cell death and MAPK signaling pathways. Conclusion: For the early COVID-19 patients with the syndrome of damp pathogen stagnation of lung, Huopo Xialing Decoction was used to resolve dampness and detoxification, and ventilate lung and promote pathogenic penetration. Phenanthrone, baicalin, jujuboside_qt, cerevisterol, hederagenin, ergosterol peroxide, citrostadienol, ergosterol-7,22-diene-3-one, paryriogenin I, alisol B,23-acetate, alisol B, neohesperidin may be the main material basis, and play a role by blocking the protein synthesis of SARS-CoV-2 virus, preventing the virus from entering the host cells, regulating the IL signaling pathway, MAPK signaling pathway, PI3K-Akt signaling pathway, T cell receptor signaling pathway, C-type lectin receptor signaling pathway and inhibiting the expression of related inflammatory factors.

Influence of alisol B 23-acetate on blood glucose in type 2 diabetic mice / 中国药理学通报

Aim To explore whether alisol B 23-acetate possesses the therapeutic potential for treatment of type 2 diabetes mellitus ( T2DM ). Methods T2DM mouse model was established by combined administration of streptozotocin and nicotinamide. After three weeks of oral administration of rosiglitazone or alisol B 23-acetate, the blood glucose of type 2 diabetic mice was measured. Oral glucose tolerance test(OGTT) was carried out the next day. Rosiglitazone was chosen as positive drug. 2-[N-(7-nitrobenz-2-oxa-l, 3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) uptake assay in adipocytes was adopted to test whether alisol B 23-acetate had effect on glucose uptake in cells. 3T3-Ll pre-adipocytes differentiation model was performed to evaluate whether alisol B 23-acetate promoted adipo-genesis. Results Mice exhibited significantly higher blood glucose concentration after intraperitoneal injection of streptozotocin and nicotinamide for three weeks, as examined by blood glucose concentration on day 21 and OGTT on day 22, compared with normal mice in blank control group. After orally administrating alisol B 23-acetate at dose of 5 mg • kg-1 , 10 mg • kg-1 ,20 mg • kg-1 daily for three weeks, respectively, or orally administered rosiglitazone at dose of 10 mg • kg-1 daily for three weeks, blood glucose greatly decreased in type 2 diabetic mice. Moreover, insulin resistance was also improved to a certain degree during OGTT. Furthermore, alisol B 23-acetate not only increased insulin-induced glucose uptake in adipocytes at the concentration of 30 mmol • L-1 glucose, but also accelerated 3T3-L1 pre-adipocytes differentiation process at concentration of 1 μmol • L-1 and 10 μmol • L-1 . Conclusions Alisol B 23-acetate reduces blood glucose of type 2 diabetic mice, promotes pre-adipocyte differentiation and increases glucose uptake in adipocytes; however, the mechanism of action needs further exploration.

Research progress of Alismatis Rhizoma and prediction analysis on its Q-marker / 中草药

Alismatis Rhizoma was first published in the “Shennong’s Classic of Materia Medica”, listed as top grade, and most of the ancient herbals have been collected. Alismatis Rhizoma is mainly distributed in Fujian, Sichuan, and Jiangxi provinces. Alismatis Rhizoma has the effect of diffusing water and dampness, releasing heat, removing turbidity, and reducing lipid. Its modern pharmacological activity is extensive, and its clinical research is deepening gradually. This paper summarizes the research status of Alismatis Rhizoma from the aspects of herbal textual research, chemical composition, and pharmacological action. On this basis, the differences of different radicals of Alismatis Rhizoma are clarified. Based on the concept of quality marker (Q-marker), the Q-markers of Alismatis Rhizoma are predicted from the point of origin, combined with the research of pharmacodynamics, new medicinal uses and processing, in order to perform qualitative and quantitative analysis of the effective components of Alismatis Rhizoma and provide scientific basis for formulation of new standards.

Preparation and quality evaluation of standard decoction of Alismatis Rhizoma / 中草药

Objective: To prepare standard decoction of Alismatis Rhizoma (AR) and establish its quality evaluation system, and provide reference for the development of dispensing granules of AR. Methods: A total of 18 batches of AR decoction pieces were collected to prepare standard decoction of AR according to the standard process. Quality evaluation system of standard decoction of AR was established with pH value, dry extract rate, fingerprint similarity and transfer rate of alisol B 23-acetate as indexes. Results: The mass fraction of alisol B 23-acetate in AR decoction pieces was 0.057%—0.267% with the average value of 0.156%, water content was 9.2%—12.8% with the average value of 10.44%; the pH value of standard decoction of AR was 4.11—5.60, dry extract rate was 10.25%—17.09%; transfer rate of alisol B 23-acetate from decoction pieces to standard decoction was 10.49%—17.49%. Conclusion: The established quality evaluation method is stable and feasible, which is suitable for the development and quality evaluation of standard decoction of AR, which can provide reference for the development of dispensing granules of AR and related classic formulas.

Exploration of objective quality evaluation parameter of Alismatis Rhizoma decoction pieces / 中国中药杂志

To establish a quality constant evaluation system of Alismatis Rhizoma decoction pieces,in order to provide reference for regulating the market circulation of this decoction pieces. A total of 18 batches of Alismatis Rhizoma decoction pieces were collected from different pharmaceutical factories,and the morphological parameters of each sample were tested. The content of alisol B 23-acetate in Alismatis Rhizoma decoction pieces was determined by HPLC in the 2015 edition of Chinese Pharmacopoeia,and the parameters such as quality constant and relative quality constant were calculated. The quality constant range of 18 batches of Alismatis Rhizoma decoction pieces was 0. 390-2. 076. If 18 batches of Alismatis Rhizoma decoction pieces were divided into 3 grades,taking 80% of the maximum quality constant as first grade,50% to 80% as second grade,and the rest as third grade,then the quality constant of firstgrade samples was ≥1. 66,the quality constant of second-grade samples was ≥1. 04 and <1. 66,and the quality constant of third-grade samples was <1. 04. The established quality constant evaluation method is objective and feasible,which can be used to classify the grade of Alismatis Rhizoma decoction pieces and provide a reference method to control the quality of this decoction pieces.

Simultaneous determination of 11 constituents in Shenqi Shiyiwei Granule by HPLC-DAD / 中草药

Objective: To establish HPLC-DAD method for the simultaneous determination of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside in Shenqi Shiyiwei Granule (SSG). Methods: The chromatographic separation was achieved on an Waters XBridge-C18 (250 mm × 4.6 mm, 5.0 μm) column with methanol-acetonitrile-water (15:80:5) and methanol-0.1% phosphoric acid (10:90) as mobile phases for gradient elution, at the flow rate of 1.0 mL/min; The column temperature was 35℃. Results: The linear ranges of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.4-8.0 μg/mL (r = 0.999 2), 0.2-4.0 μg/mL (r = 0.999 5), 0.2-4.0 μg/mL (r = 0.999 5), 0.1-2.0 μg/mL (r = 0.999 6), 0.1-2.0 μg/mL (r = 0.999 7), 0.1-2.0 μg/mL (r = 0.999 4), 0.5-10 μg/mL (r = 0.999 2), 0.6-12 μg/mL (r = 0.999 2), 0.4-8.0 μg/mL (r = 0.999 4), 1.0-20 μg/mL (r = 0.999 6), and 0.8-16 μg/mL (r = 0.9993). The average recoveries (n = 6) were 98.1% (RSD = 0.9%), 98.1% (RSD = 1.6%), 99.1% (RSD = 1.6%), 98.3% (RSD = 1.8%), 99.5% (RSD = 1.5%), 99.9% (RSD = 0.6%), 98.5% (RSD = 0.7%), 100.4 (RSD = 0.8%), 101.6% (RSD = 0.4%), 99.7% (RSD = 0.9%), and 101.2% (RSD = 1.1%), respectively. The contents of nine batches of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.081-0.089, 0.261-0.269, 0.060-0.069, 0.038-0.047, 0.030-0.037, 0.042-0.049, 0.420-0.428, 0.141-0.151, 0.178-0.189, 0.107-0.117, and 0.069-0.078 mg/g. The results showed that there was little difference among the batches. Conclusion: The method is accurate, sensitive, credible, and repeatable. It can be applied to the quality control of SSG.

Optimization of flash-type extraction technology of alisol B 23-acetate from Alismatis Rhizoma by response surface methodology / 中国中药杂志

Response surface methodology was used to optimize and obtain the optimal flash-type extraction technology of alisol B 23-acetate from Alismatis Rhizoma. With the extraction rate of alisol B 23-acetate as an indicator, single-factor test was used to investigate the effect of ethanol volume fraction, liquid-solid ratio, extraction times and extracting time on the extraction rate of alisol B 23-acetate.The results were combined with Box-Benhnken design and response surface analysis to optimize the technology parameters for extraction process of Alismatis Rhizoma and obtain the optimal flash-type extraction technology under the following conditions： ethanol volume fraction 80%, liquid-solid ratio 12∶1, extraction 4 times, 114 s/time. Flash-type extraction technology of alisol B 23-acetate by response surface methodology is stable, time-saving, efficient, and with the advantages of room temperature extraction and no component damage, so it can be used for massive production.

Research on effective components compatibility of Alismatis Rhizoma resisting calcium oxalate calculus in vitro based on uniform design / 中草药

Objective: To investigate the inhibitory effect of six active triterenoids and their compatibility from Alisma orientale on the formation of urinary calcium oxalate calculus in vitro. Methods: A uniform design method was used to design the different compatibilities from six triterenoids. The inhibitory effects of the six triterenoids along with their different compatibility groups were evaluated in vitro using a standard seeded crystallization technique. Results: The six active triterenoids and their compatible preparations could significantly inhibit calcium oxalate crystal formation in vitro (P < 0.05), and the group of alisol A-alisol A 24-acetate-alisol B-alisol B 23-acetate-alisol F-alisol F 24-acetate (2.2∶3.8∶1∶3.5∶2.2∶1) was the most efficient with an inhibitory index of 188.29%. Conclusion: The triterenoids in A. orientale play a key role in the inhibition of calcium oxalate calculus formation and compatibility of alisol A, alisol A 24-acetate, alisol B, alisol B 23-acetate, alisol F, and alisol F 24-acetate can inhibit the calcium oxalate calculus well in vitro especially cooperated with each other, and the best compatibility proportion is 2.2∶3.8∶1∶3.5∶2.2∶1, respectively.

Simultaneous determination of 11 triterpenoids in Alismatis Rhizoma by RP-HPLC-DAD / 中草药

Objective: To establish a RP-HPLC-DAD method for simultaneous determination of alisol C, alisol F, alisol C 23-acetate, alisol L, alisol F 24-acetate, alisol A, alisol A 24-acetate, alisol G, alisol B, alisol B 23-acetate, and 11-deoxy-alisol B in Alismatis Rhizoma. The content of 11 components from 25 batches of samples collected from different habitats was determined and compared by means of this established method. Methods: The separation was achieved on a Ultimate XB-C18 column (150 mm × 4.6 mm, 5 μm) with the mobile phase consisting of water-acetonitrile to be gradient elution. Flow rate was 1 mL/min and column temperature was 30℃. The optimum detection wavelengths were 208 and 245 nm. Results: The linear ranges of alisol C, alisol F, alisol C 23-acetate, alisol L, alisol F 24-acetate, alisol A, alisol A 24-acetate, alisol G, alisol B, alisol B 23-acetate, and 11-deoxy-alisol B were 0.313-17.210 (r = 0.999 9), 0.504-11.880 (r = 0.999 9), 0.284-9.369 (r = 0.999 9), 0.146-2.680 (r = 0.999 9), 0.254-5.596 (r = 0.999 8), 2.500-66.000 (r = 0.999 8), 1.620-35.640 (r = 0.999 9), 1.575-20.475 (r = 0.999 9), 1.525-57.268 (r = 0.999 6), 3.035-118.362 (r = 0.999 9), and 0.816-16.880 μg/mL (r = 0.999 8), respectively. The average recoveries (n = 6) were 98.76%, 100.24%, 97.97%, 100.14%, 99.88%, 96.54%, 97.27%, 98.85%, 99.45%, 98.85%, and 98.13% respectively. Conclusion: The RP-HPLC-DAD method is feasible and accurate, which could provides the scientific evidence for quality control of Alismatis Rhizoma.

Preparation process of lipid-lowering medicines by solid state fermentation of Crataegi Fructus, Alismatis Rhizoma, and Cassiae Semen mixed with monascus / 中草药

Objective: To obtain a new lipid-lowering product that combines the advantages of lipid-lowering Chinese materia medica (CMM) and statin drugs. Solid-state fermentation was studied using medicinal and edible CMM as media components, Lovastatin yielding monascus screened in our laboratory was used as strain and Lovastatin yielding was used as index. Methods: Solid-state fermentation was optimized by single factor and orthogonal test, qualitative and quantitative analysis was conducted of the product via TLC and HPLC. Results: The optimal fermentation conditions contained 100 g/solid medium in 500 mL flask bottling capacity, material thickness of 2.5 cm, 10 mL liquid seed of 48 h, 10% inoculation volume, incubating at 30℃, breaking up the medium on day 2, adding 24% sterile water on day 3, cooling to 25℃ and culturing for 18 d in total, and Lovastatin yielding was up to 5.127 mg/g. Fermentation product of CMM contained more components compared with that without medicines, Lovastatin yielding increased by 42.27%, but γ-aminobutyric acid yielding decreased by 17.89%. Contents of main active ingredients ursolic acid, 2,3-acetyl alisol-B, chrysophanol, and physcion were increased by 232.7%, 173.7%, 767.6%, and 888.4%. Conclusion: Active ingredients of hawthorn, alisma, and cassia are released into the products after fermentation and contents of lipid-lowering active ingredients were improved significantly, new active ingredients are also noticed. Therefore, the fermentation process of lipid-lowering medicines obtained in this experimental study has some practical values.

Simultaneous determination of eleven ingredients in Xinnaokang Tablets by HPLC / 中草药

Objective: To establish a method of HPLC for thesimultaneous determination of eleven ingredients (tanshinone IIA, puerarin, β-ecdysone, liquiritin, paeoniflorin, spinosin, ferulic acid, kaempferide, emodin, tenuifolin, and 23-acetate alisol B) in Xinnaokang Tablets. Methods: The contents of tanshinone IIA, puerarin, β-ecdysone, liquiritin, paeoniflorin, spinosin, ferulic acid, kaempferide, emodin, tenuifolin, and 23-acetate alisol B were determinedby HPLC. Separation was performed on an InertSustain C18column (250 mm × 4.6 mm, 5 μm); Mobile phase was methanol-acetonitrile (1∶1, A)-0.1% phosphoric acid (B), gradient elution: 0-10.0 min, 80% B; 10.0-20.0 min, 80%-70% B; 20.0-35.0 min, 70%-50% B; and 35.0-50.0 min, 50%-30% B; Flow rate was 0.9 mL/min; Column temperature was 40 ℃; Injection volume was 10 μL. Results: The separations of tanshinone IIA, puerarin, β-ecdysone, liquiritin, paeoniflorin, spinosin, ferulic acid, kaempferide, emodin, tenuifolin, and 23-acetate alisol B were good.The linear ranges were good,28.24-282.42, 15.89-158.90, 20.53-205.26, 4.09-40.94, 12.08-120.76,40.03-404.30, 4.08-40.75, 2.13-21.25, 7.91-79.14, 20.30-202.96, and 4.10-40.96μg/mL for tanshinone IIA, puerarin, β-ecdysone, liquiritin, paeoniflorin, spinosin, ferulic acid, kaempferide, emodin, tenuifolin,and 23-acetate alisol B, respectively, with the correlation r> 0.999 0. The extraction recoveries varied from 98.15% to 101.84% (RSD varied from 0.62% to 1.71%). The contents of tanshinone IIA, puerarin, β-ecdysone, liquiritin, paeoniflorin, spinosin, ferulic acid, kaempferide, emodin, tenuifolin,and 23-acetate alisol Bwere 0.870-0.887, 6.321-6.329, 0.857-0.866, 0.171-0.179, 0.369-0.377, 2.128-2.136, 0.088-0.099, 0.148-0.155, 0.113-0.121,0.371-1.380, and 0.101-0.109 mg/gin six batches of samples, respectively. Conclusion: This method is rapid and has high sensitivity, high accuracy, and good specificity,which can provide the basis for the quality control of Xinnaokang Tablets.

Comparison on six active constituents between formula granule and traditional decoction of Rehmanniae Decoction of six ingredients / 中草药

Objective: To compare the content of chemical components in each drug in formula granule and traditional decoction of Rehmanniae Decoction with six ingredients. Methods: By using high performance liquid chromatography analysis and various chromatographic conditions, the contents were determined respectively, which were listed as follows: acteoside, loganin, paeonol, allantoin, pachymic acid and alisol B 23-acetate. Results: The contents of acteoside, loganin, paeonol, allantoin, pachymic acid and alisol B 23-acetate in traditional decoction were 304.5, 2 473.6, 3 135.1, 708.8, 5.9, and 104.4 μg/g, and they were 289.6, 3 685.7, 706.5, 714.2, 17.4, and 217.8 μg/g correspondingly in formula granule. The contents of acteoside and allantoin were basically the same between them; The contents of loganin, pachymic acid, and alisol B 23-acetate in formula granule were significantly higher than those in the traditional decoction; The content of paeonol in formula granule was significantly lower than that in the traditional decoction. Conclusion: The content difference of the chemical components is related to its chemical character between formula granule and traditional decoction.

Comparison on Contents of 23-Acetate Alisol B and Infrared Spectrometry Fingerprint Among Rhizoma Alismatis of Different Specifications / 中国中医药信息杂志

Objective To compare the chemical components among Rhizoma Alismatis of different specifications. Methods Rhizoma Alismatis of 8 different weights were chosen, and then contents of 23-acetate alisol B were determined by HPLC, and infrared spectrometry fingerprint was determined by Fourier transform infrared spectroscopy. Results The contents of 23-acetate alisol B in Rhizoma Alismatis of 8 different specifications were over 0.06%, and had no relation with specification of Rhizoma Alismatis (P>0.05). The similarities of infrared spectrometry fingerprint were above 0.9. Conclusion The chemical components among Rhizoma Alismatis of different specifications were basically the same. Contents of 23-acetate alisol B of Rhizoma Alismatis of 8 different specifications conformed to regulation of China Pharmacopoeia.

Extraction and isolation of 23-acetate alisol B from Alisma orientalis by supercritical fluid extraction and high-speed counter-current chromatography / 中草药

Objective: To develop an effective and rapid method for the preparation of 23-acetate alisol B from Alisma orientalis. Methods: The SFE-CO2 extract from A. orientalis was injected into high speed counter current chromatography (HSCCC) directly, and eluted with difierent solvent systems. The crystalline purity was detected by HPLC. The structure of the target compound was identified by UV, IR, MS, and NMR. Results: The solvent system composed of n-hexane-ethylacetate- methanol-water (3∶2∶3∶2) was the best. The lower phase was used as the mobile phase and performed at a flow rate of 2 m/min, while the apparatus rotated at 800 r/min, and detected at 254 nm. The prepared alisol B 23-acetate was identified with infrared spectrometry (IR), mass spectrometry (MS), and nuclear magnetic resonance (NMR) detection, and its purity was 99.8% analyzed by HPLC. Conclusion: The established method is relatively simple, fast, and suitable for the fast isolation and separation of alisol B 23-acetate.

Chemical constituents from Alisma orientalis extracts with hypoglycemic effect / 中草药

Objective: To study the chemical constituents in Alisma orientalis extracts with hypoglycemic effect. Methods: To study the in vivo hypoglycemic effects of A. orientalis extracts, high fat diet (HFD)-induced insulin resistance male C57BL/6J mice were treated with water and ethanol extracts of A. orientalis in diet, and glucose tolerance test was carried out following the intervention. Silica gel, ODS, and preparative HPLC were used to isolate the compounds. Their chemical structures were elucidated on the basis of NMR and MS spectral data. Results: Sixteen compounds were identified as sitosterol (1), palmitic acid (2), heptadecanoic acid (3), eicosanoic acid (4), 11-deoxy-alisol B (5), 23-acetate alisol B (6), 23-acetate alisol C (7), alisol B (8), 24-acetate alisol A (9), alisol G (10), 24-acetate alisol F (11), alisol L (12), alisol C (13), alisol F (14), alisol A (15), and 16-oxo-24-acetate alisol A (16), and nine of the triterpenes could improve glucose uptake in HepG2 cells. Conclusion: Compounds 3 and 4 are isolated from A. orientalis for the first time. The water and ethanol extracts of A. orientalis could improve glucose tolerance test. Triterpenes may be one of the therapeutic material basis in hypoglycemic activities in A. orientalis.

Determination of 23-Acetate Alisol B in Longdan Xiegan Honey Pills by HPLC / 中国药师

Objective:To establish a method for the determination of 23-acetate alisol B in Longdan Xiegan honey pills by HPLC. Methods:The analysis was performed on a Waters Symmetry C18 (250 mm × 4. 6 mm,5μm) column with the mobile phase of acetonitrile-0. 1% phosphoric acid (62 ∶ 38). The flow rate was 1. 0 ml·min-1, the column temperature was 35℃ and the detection wavelength was set at 208nm. Results: The linear range of 23-acetyl alisol B was 19. 999 5- 1 999. 9500 ng(r =0. 999 9), and the average recovery was 95. 56%(RSD = 0. 7%, n = 6). Conclusion: The method is simple, rapid and accurate, and can be used to control the quality of Longdan Xiegan honey pills with good repeatability and recovery.

Determination of cinnamic acid, cinnama ldehyde, paeonol, alisol B 23-acetate, atractylodes lancea and atractylodin in Yeming granules by RP-HPLC / 中国药学杂志

OBJECTIVE: To develop a RP-HPLC method for determination of cinnamic acid, cinnama ldehyde, paeonol, alisol B 23 acetate, atractylodes lancea, and atractylodin in Yeming granules. METHODS: The determination was performed on an Eclipse XDB-C18column (4.6 mm × 250 mm, 5 μm) at 30°C with a flow rate of 1.0 mL · min-1. The mobile phase was 0.3% phosphate-acetonitrile at gradient elution. The detection wavelength was set at 240 nm. RESULTS: The linear regression equations for cinnamic acid, cinnama ldehyde, paeonol, alisolB 23 acetate, atractylodes lancea, and atractylodin were ρ=-0.0055A + 0.0083 (r=0.9996), ρ=0.0255, 4+0.1336 (r=0.9999), ρ=0.0408 A +0.664 3 (r=0.999), ρ=0.003 3 4+0.0171(r=0.999 9), ρ=0.0163 A + 0.1075 (r=0.999 8), ρ=0.0229A + 0.0884 (r=0.9999), respectively. Good linearity was obtained over the ranges of 0.3096-2.322, 10.24-76.8, 37.36-280.2, 0.5128-3.846, 1.612-12.09 and 10.26-76.95 μg · mL-1, respectively. The recoveries were 100.1%, 99.8%, 100.7%, 99.3%, 100.4% and 100.5%, respectively. CONCLUSION: The method is simple and suitable to control the quality of Yeming granules. Copyright 2012 by the Chinese Pharmaceutical Association.

Study on extraction technology of Zhixuan Granula / 中草药

Object To optimize the preparation procedure for Zhixuan Granula (ZXG). Methods The optimum extracting conditions of ZXG were selected by orthogonal test with the active components: 23-acetate alisol B, atractylenolide I, and dried extract as the index, it mice sedation of ZXG was clarified by pharmacodynamics. Results The optimum preparation procedure was as follows: Rhizoma Alismatis and Rhizoma Atractylodis Macrocephalae were extracted with alcohol first, adding 12-fold 70% alcohol by refluxing, extracting twice, 2 h once, then extracted with water, adding 14-fold water, extracting twice, 2 h once. The extract showed the obvious effect on sedation of mice. Conclusion The optimum preparation procedure is reliable, with higher extracting ratio of the active components.

Transformation of alisol B 23-acetate in processing of Alisma orientalis / 中草药

Objective To study the transformation mechanism of triterpenes in processing of Alisma orientalis. Methods The triterpene transformations of A. orientalis pre and post-processing were comparatively analyzed by techniques of HPLC and Packed Column Supercritical Fluid Chromatography (SFC). Results In baked processing (70 ℃) of A. orientalis, little alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B.However, more alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B, then both of them were further transformed into alisol A in processing under high temperature (160-200 ℃). Conclusion Transformation of alisol B 23-acetate has two routes when A. orientalis is processed under high temperature: For one, alisol B 23-acetate is rearranged into alisol A 24-acetate which could be deacetylated into alisol A; for the other; it is deacetylated into alisol B first, then transformed into alisol A.