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Objective: To investigate the therapeutic effect and potential mechanism of Zhenwu Decoction (ZWD) on chronic heart failure (CHF) rats. Methods: HPLC fingerprint of ZWD was established. All male SD rats were randomly divided into the sham operation group, the model group, the low, medium and high dose ZWD group (2.187 5, 4.375, and 8.75 g/kg) and the captopril group (10 mg/kg). Except for the sham operation group, the rest of rats were all established into the CHF model rats by ligating the left anterior descending branch of the coronary artery, after 8 weeks, all rats were ig administration for 4 weeks. The hemodynamic, viscera index, HE dyeing test were conducted at the end of experiments. Serum angiotensin II (Ang II), aldosterone (ALD), nuclear factor kappa B (NF-κB), amino terminal brain natriuretic peptide (NT-proBNP), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) were determinated by ELISA, and myocardial NF-κB protein expression was detected by Western blotting. Results: Higenamine, paeoniflorin, atractylenolide III, 6-gingerol and dehydrotumulosic acid, the five constituents of Zhenwu Decoction, were identified by HPLC chart. Compared with the model group, the administration of the ZWD significantly improved the hemodynamic parameters (P < 0.05), reduced the organ index (P < 0.05) and improved myocardial injury, reduced the serum Ang II, ALD, NF-κB, NT-proBNP, TNF-α and IL-6 levels and the myocardial NF-κB protein expression (P < 0.05). Conclusion: HPLC results provided an evidence for the quality control and pharmacodynamic substance of ZWD. ZWD can ameliorate CHF, which may be related to the inhibition of renin- angiotensin-aldosterone system (RAAS)/NF-κB/inflammatory factor cascade reaction.
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Objective: To construct the fingerprint of the triterpenoic acid in Guizhi Fuling Capsule (GFC) by UPLC/Q-TOF-MS, and give a new method for its quality control. Methods: The triterpenoic acid were isolated by UPLC and detected by Q-TOF- MS. The established fingerprint was an atlas of UPLC/Q-TOF-MS. Results: The UPLC/Q-TOF-MS fingerprints of triterpenoids in GFC were established, which preserved high precision, selectivity, and specificity. A total of 26 common peaks were selected as the fingerprint peaks of GFC, of which a total of 18 mutual peaks (3, 5-18, 20, 23, and 24) from Poria, peak 2 was from Paeoniae Radix Alba and Moutan Cortex. Peak 4 was from Poria, Moutan Cortex, Paeoniae Radix Alba, and Cinnamomi Ramulus. Peak 19 was from Moutan Cortex, Paeoniae Radix Alba, and Cinnamomi Ramulus, peak 21 was from Moutan Cortex and Paeoniae Radix Alba, peaks 22 and 25 were from Poria, Moutan Cortex, Persicae Semen, Paeoniae Radix Alba, and Cinnamomi Ramulus, peak 26 was from Cinnamomi Ramulus, Paeoniae Radix Alba, and Persicae Semen. The similarity among the 10 batches of GFC was above 0.90. The Results: of validation met technical requirement of fingerprints. Sixteen chemical components were identified by UPLC-Q-TOF-MS, which were 16α-hydroxydehydrotrametenolic acid, 16α-hydroxychalcic acid, 3-oxo-6,16α-dihydroxy-lanosta-7,9(11),24(31)-trien-21- oic acid, dehydrotumulosic acid, tumulosic acid, 3-oxo-6,16α-dihydroxy-lanosta-8,24-diene-21-oic acid, eburicoic acid, polyporenic acid C, 3-epidehydrotumulosic acid, 3-O-acetyl-16α-hydroxydehydrotrametenolic acid, 3-epidehydropachymic acid, 3-O-acetyl-16α- hydroxyplugymic acid, dehydropachymic acid, pachymic acid, dehydrotrametenolic acid, and dehydroeburicoic acid. Conclusion: The method is rapid, accurate, and has desirable precision, reproducibility, and stability, which could be applied to the quality control of GFC.
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Objective To establish a new method and validate its feasibilities by the simultaneous quantitative assay of four triterpenes in Poria cocos. Methods A new quality evaluation method of multi-components by single marker (QAMS) was established and validated for P. cocos. Pachymic acid (PA), dehydropachymic acid (DPA), dehydrotumulosic acid (DTA), and dehydrotrametenolic acid (DMA) were selected as analytes while pachymic acid was chosen as internal reference substance to evaluate the quality. The relative correction factor (RCF) of pachymic acid to the other three triterpenes were calculated. The method was evaluated by the comparison of quantity between external standard method and QAMS method. Results The contents of four triterpenes in 17 batches of P. cocos from QAMS method were not significantly different from those from external standard method. Conclusion The method with a single marker is accurate and feasible to determine PA, DPA, DTA, and DMA when some authentic standard substance are unavailable.
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Objective: To establish the UPLC fingerprint of triterpenoids in Guizhi Fuling Capsule (GFC) and give a new method for quality control. Methods: UPLC was used on an Agilent Zorbox Eclipse Plus HD C18 (100 mm × 2.1 mm, 1.8 μm) column with the gradient elution solvent system composed of acetonitrile-0.1% H3PO4 water solution as mobile phase, the flow rate was 0.2 mL/min, the column temperature was 30℃, and the detection wavelength was 210 nm. The common peaks were identified by Q-TOF/MS. Results: The UPLC fingerprints of triterpenoids in GFC were established. Totally 20 common peaks were selected as the fingerprint peaks of GFC, of which a total of 13 mutual peaks (1-7, 10, 12, and 14-17) from Poria, the peaks of number 8 and 11 were from Cinnamomi Ramulus, the peak of number 9 was from Paeoniae Radix Alba and Moutan Cortex, and the peak of number 13 was from Persicae Semen, Paeoniae Radix Alba, Moutan Cortex, and Cinnamomi Ramulus. The similarity among the fingerprint of 10 batches of GFC samples were 0.90. Fifteen chemical components were identified by UPLC-Q-TOF/MS, which were 1-hederagenin, 2-dehydrotumulosic acid, 3-tumulosil acid, 4-polyporenic acid C, 6-3-epidehydrotumulosic acid, 7-poricoic acid D, 9-α-linolenic acid, 10-dehydropachymic acid, 11-oleanolic acid, 12-pachymic acid, 13-linolic acid, 15-methylcis-9-hexadecenoate, 16-palmitic acid, 17-palmitic acid ethyl ester, and 18-daucosterol. Conclusion: The established UPLC fingerprint has desirable precision, reproducibility, and stability, and could be applied to the quality control of GFC.
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Objective: To establish an HPLC method for the determination of four triterpene constituents (dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydropachymic acid, and dehydropachymic acid) in Guizhi Fuling Capsules (GFC). Methods: Chromatography conditions were Diamonsic C18 column (250 mm × 4.6 mm, 5 μm), system mobile phase was composed of acetonitrile (A)-0.2% HCOOH aqueous solution (C) in a linear gradient elution mode (0-70 min: 50%-85% A; 70-80 min: 85%-100% A; 80-90 min: 100% A), detective wavelength was set at 242 nm, and column temperature was 40℃. Results: The calibration curve was linear within 0.408-2.04 μg/mL (r = 0.999 9), 0.192-0.96 μg/mL (r = 0.999 5), 0.078-0.39 μg/mL (r = 0.999 5), and 0.075 6-0.378 μg/mL (r = 0.999 5) for dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydropachymic acid, and dehydropachymic acid, respectively. The average recoveries were 97.5% (RSD = 1.4%, n = 5), 98.5% (RSD = 1.6%, n = 5), 97.2% (RSD = 1.2%, n = 5), and 102.3% (RSD = 1.8%, n = 5). Six batches of GFC sample were determined, The average contents of dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydropachymic acid, and dehydropachymic acid were 0.070, 0.015, 0.030, and 0.061 mg/capsule, separately. Conclusion: The method is simple, accurate, and can be used as a quality control method for GFC.
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To establish a method for directional separation of dehydrotumulosic acid from the extracts of Guizhi Fuling Capsule with molecular imprinting technique (MIT). Molecular imprinting polymer (MIP) was prepared by sol-gel process with dehydrotumulosic acid as molecular template to study the absorption property. The dehydrotumulosic acid was achieved from Guizhi Fuling Capsule by one-step separation with polymer as filler. The structure of dehydrotumulosic acid was identified on the basis of the spectral data and physicochemical property. The maximum binding capacity (Qmax) of MIP was 9.10 mg/g measured by Scatchard equation and the purity of dehydrotumulosic acid was 90.76% by HPLC. The established MIT for the directional separation of dehydrotumulosic acid from Guizhi Fuling Capsule is simple and benefit to reducing the solvent use during the separation process, which could offer a novel method for the separation and purification of dehydrotumulosic acid.
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OBJECTIVE: To establish an analytical method to determine dehydrotumulosic acid, tumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid and pachymic acid in poria quickly and accurately. METHODS: An UPLC method was established on an HSS T3 Column (2.1 mm × 100 mm, 1.8 μm). The mobile phase consisted of water (containing 0.05% phosphoric acid) and acetonitrile in gradient elution mode, and the detection wavelength was set at 210 nm. RESULTS: The standard curves of dehydrotumulosic acid, tumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid, and pachymic acid showed good linearity in 5.400-108.0, 2.040-40.80, 5.020-100.4, 2.120-42.40, 5.060-101.2 and 5.100-102.0 μg · mL-1, respectively and the average recoveries were 98.0% with RSD of 2. 9% for dehydrotumulosic acid, 99. 0% with RSD of 2.8% for tumulosic acid, 101.5% with RSD of 2.5% for polyporenic acid C, 97.1% with RSD of 2.7% for 3-epi-dehydrotumulosic acid, 101.5% with RSD of 2.1% for dehydropachymic acid and 99.6% with RSD of 1.1% for pachymic acid, respectively. CONCLUSION: The method is quick, accurate, and can be used to determine multiple triterpenoid acids in Poria simultaneously.