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1.
Article in Chinese | WPRIM | ID: wpr-906217

ABSTRACT

Objective:Through comprehensive evaluation and analysis of the quality of Liuwei Dihuang (LWDH) preparations from different manufacturers and combining factors such as production technology, the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations. Method:Morroniside, loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A) -0.3% phosphoric acid aqueous solution (B) for gradient elution (0-5 min, 5%-8%A; 5-20 min, 8%A; 20-35 min, 8%-20%A; 35-45 min, 20%-60%A; 45-55 min, 60%A), the detection wavelength of paeonol was at 274 nm, and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms (big candied pills, water-honeyed pills, concentrated pills, hard capsules and soft capsules) from different manufacturers were analyzed. Combined these results with their actual production processes, the key-points of quality control in the whole production process were discussed. Result:The contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of <italic>Chinese Pharmacopoeia</italic>, however, the content limit of some dosage forms in the current standard was unreasonable. For example, although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills (15.00, 8.57 g, respectively), they got exactly the same daily limits of the contents for both the quality markers. What′s more, these two formulations had the same process, so the differences between the process obviously could not be the reason of these differences. Conclusion:It is recommended that for the products with different dosage forms should have a similar content limits, if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards, and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.

2.
Article in Chinese | WPRIM | ID: wpr-846568

ABSTRACT

Objective: To screen the differential ingredients between crude and wine-processed Corni Fructus and determin their content. Methods: An integrated strategy using ultra performance liquid chromatography coupled with tandem quadrupole time-of- flight mass spectrometry (UPLC-Q-TOF/MS) and the chemometric approach was applied to compare the global chemical profile of crude and wine-processed Corni Fructus. Then, the main differential ingredients were quantified by UPLC-PDA. Results: The chemical profiling of wine-processed Corni Fructus was significantly different. Ten compounds could be considered as characteristic chemical markers for distinguishing crude and wine-processed Corni Fructus, including 5-hydroxymethyl furfuraldehyde (5-HMF), gallic acid, protocatechuic acid, morroniside, loganic acid, sweroside, cornin, dihydroquercetin, loganin and cornoside. A new UPLC-PDA quantitative method for analyzing simultaneously the above ten compounds in wine-processed Corni Fructus was established. The results of methodology investigation showed that the ten components were well linear within the investigation range (r ≥ 0.999 7). Compared with the crude Corni Fructus, the content of seven components were increased, including gallic acid, 5-HMF, loganin, morroniside, cornin, sweroside and dihydroquercetin, and the other three components in wine-processed Corni Fructus were decreased. Conclusion: The differential ingredients obtained by chemometric-based approach can be used to distinguish crude and wine-processed Corni Fructus. The determination method of wine-processed Corni Fructus established is accurate and reliable, which can be used for the quality control of Corni Fructus.

3.
Article in Chinese | WPRIM | ID: wpr-846388

ABSTRACT

Objective: To establish an HPLC fingerprint of Qingxin Zishen Prescription Decoction (QZPD) and determine the contents of its multiple components, so as to provide a scientific basis for quality control. Methods: HPLC analysis was performed on a Phenomenex Kinetex C18 column (100 mm × 4.60 mm, 2.6 μm) for gradient elution with the mobile phase consisting of methanol, acetonitrile and 0.2% formic acid aqueous. The detection wavelength was set at 245 nm and 280 nm, and the column temperature was 40 ℃. Fingerprints of ten batches of QZPD were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed and the contents of 15 components were determined. Results: The fingerprint similarities of 10 batches of QZPD were ranged from 0.923 to 0.998 compared with the reference fingerprint, and 33 common peaks were identified in the fingerprint. Among them, seven peaks (P11, P14-P16, P24, P29, P30) were identified from Coptidis Rhizoma, two peaks (P7, P19) were identified from Nelumbinis Plumula, two peaks (P14, P21) were identified from Ziziphi Spinosae Semen, nine peaks (P4, P5, P10, P17, P18, P28, P31-P33) were identified from Salvia miltiorrhiza, nine peaks (P1-P3, P6, P8, P9, P12, P13, P20) were identified from Corni Fructus, while five peaks (P22, P23, P25-P27) cannot be originated and none of the common peaks was identified from Rehmanniae Radix, Uncariae Ramulus Cum Uncis and Triticum aestivum. By comparing with the chemical reference, fifteen components, including gallic acid (P2), 5-hydroxymethylfurfural (P3), danshensu (P4), protocatechuic aldehyde (P5), morroniside (P9), caffeic acid (P10), cornin (P12), loganin (P13), magnoflorine (P14), coptisine (P24), lithospermic acid (P28), berberine (P29), palmatine (P30), salvianolic acid B (P31) and salvianolic acid E (P33), were identified and quantified. The contents of the fifteen components were 158.3-248.2, 233.6-321.3, 45.9-166.0, 24.3-38.6, 800.7-1 263.6, 26.6-54.9, 44.5-108.2, 470.4-757.3, 85.6-178.6, 11.1-34.2, 56.2-106.4, 25.9-138.9, 21.0-59.2, 951.6-2 244.7 and 38.6-92.8 μg/g, respectively. Conclusion: The method established in this study is stable and highly reproducible, and can provide basis for quality control of QZPD.

4.
China Pharmacy ; (12): 2508-2511, 2020.
Article in Chinese | WPRIM | ID: wpr-829359

ABSTRACT

OBJECTIVE:To estab lish a me thod for simultaneous determination of morroniside ,loganin,echinacoside and acteoside in Huanshao capsules. METHODS :HPLC method was adopted. The determination was performed on Zhongpuhong RD-C18 column with mobile phase consisted of acetonitrile- 0.1% formic acid solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm (morroniside,loganin) and 330 nm (echinacoside,acteoside). The column temperature was set at 35 ℃,and sample size was 10 μL. RESULTS:The linear range were 5.29-105.80 μg/mL for morroniside, 4.49-89.88 mg/L for loganin ,16.26-325.25 mg/L for echinacoside and 16.31-326.25 mg/L for acteoside ,r values were 0.999 9. RSDs of precision ,stability (24 h),reproducibility and durability tests were all lower than 2.0% . The recoveries were 94.34% -96.23%(RSD=0.81% ,n=6),97.04% -98.89%(RSD=0.73% ,n=6),96.23% -98.08%(RSD=0.82% ,n=6), 95.40%-98.47%(RSD=1.23%,n=6),respectively. The contents of above 4 components in 11 batches of Huanshao Capsules were 0.190-0.704,0.439-0.857,2.723-4.475 and 0.589-1.035 mg/g,respectively. CONCLUSIONS :Established method is specific , precise and can be used for content determination of 4 components in Huanshao capsules.

5.
Article in Chinese | WPRIM | ID: wpr-851334

ABSTRACT

Objective: To establish a rapid, accurate, and practical HPLC method for simultaneous determination the content in Qiju Dihuang Oral Liquid (QDOL) of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and paeonol. Methods: YMC ODS column (250 mm × 4.6 mm, 5 μm) was used, column temperature was set at 35 ℃, gradient elution with 0.1% formic acid aqueous solution- acetonitrile was used as mobile phase, flow rate was 1.0 mL/min, detection wavelength was 254 and 325 nm. The injection volume was 10 μL. Results: The injection amount of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, paeonol injection quality at 0.08—1.60, 0.12—2.40, 0.09—1.80, 0.06—1.20, 0.10—2.00, 0.30—6.00, 0.01—0.20, 0.01—0.20, 0.01—0.20, 0.005—0.10, 0.005—0.10, and 0.01—0.20 μg showed a good linear relationship with peak area, with good precision, repeatability and stability. The recovery rates of the samples were between 96% and 103%, the RSD was 2.13%, 3.45%, 2.86%, 2.59%, 3.15%, 3.49%, 2.19%, 3.25%, 2.37%, 2.53%, 2.91%, and 3.35%, respectively. The content of each component of the five batches of samples was stable, and the mass concentrations range of the 12 components tested were 98.56—102.56, 204.28—212.10, 18.53—18.89, 1.95—2.05, 12.31—12.54, 87.01—87.12, 5.35—5.43, 16.08—16.15, 8.69—8.72, 8.89—8.95, 5.12—5.19, and 1.87—1.94 μg/mL. Conclusion: The method simltaneosly determines the content of 5-HMF, morroniside, chlorogenic acid, cryptochlorogenic acid, loganin, paeoniflorin, verbascoside, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and paeonol in QDOL, which is suitable for the quality control of QDOL.

6.
Article in Chinese | WPRIM | ID: wpr-851059

ABSTRACT

Objective: To compare the differences in pharmacokinetic behavior of six ingredients in Qikui Sustained-release Tablets in rabbit plasma. Qikui Granules was taken as reference. Methods: Diazepam was used as internal standard. LC-MS/MS detection methods of astragaloside, hyperin, isoquercitrin, rutin, morroniside, and loganin in rabbit plasma were established, and pharmacokinetic parameters of six components were calculated. Results: Six active ingredients’ equation of linear regressions were: astragaloside Y = 1.0 × 10-4 X - 0.009 9 (r = 0.999 7), morroniside Y = 1.0 × 10-4 X + 0.038 7 (r = 0.999 4), loganin Y = 3.0 × 10-5 X + 0.008 7 (r = 0.999 3), hyperin Y = 1.0 × 10-3 X - 0.016 1 (r = 0.999 0), rutin Y = 5.0 × 10-4 X - 0.011 5 (r = 0.999 4), isoquercitrin Y = 1.7 × 10-3X - 0.307 5(r = 0.999 2). Intra-day and inter-day precision and accuracy and recovery rate were up to the mustard. After Qikui Sustained-release Tablets and Qikui Granules being given by gavege, the maximal concentration (Cmax) of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Granules were (1.333 ± 0.051), (1.238 ± 0.164), (0.83 ± 0.079), (0.127 ± 0.017),(0.444 ± 0.048), and (0.223 ± 0.048) mg/L, t1/2 were (3.848 ± 0.311), (3.822 ± 0.757), (4.982 ± 1.14), (3.73 ± 0.298), (4.732 ± 0.642), and (5.132 ± 0.901) h, respectively, AUC(0-t) were (3.069 ± 0.307), (2.891 ± 0.943), (2.079 ± 0.306), (0.313 ± 0.068), (1.087 ± 0.177), (0.496 ± 0.129) mg∙h/L, respectively, Cmax of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Sustained-release Tablets were (0.985 ± 0.13), (0.961 ± 0.175), (0.693 ± 0.101), (0.094 ± 0.012), (0.354 ± 0.045), (0.201 ± 0.037) mg/L, t1/2 were (4.691 ± 0.337), (5.62 ± 1.64), (6.408 ± 0.707), (4.103 ± 0.341), (6.048 ± 0.882), (5.803 ± 0.59) h, AUC(0-t) were (5.191 ± 1.046), (6.168 ± 1.25), (4.293 ± 0.823), (0.485 ± 0.103), (1.84 ± 0.432), (0.924 ± 0.19) mg∙h/L. Contrast with Qikui Granules, relative bioavailability of morroniside, loganin, astragaloside, rutin, hyperin, and isoquerctirin in Qikui Sustained-release Tablets were 169.1%, 213.3%, 206.5%, 156.0%, 169.3%, and 186.3%, respectively. Conclusion: Qikui Sustained-release Tablets can significantly improve the bioavailability of each active ingredient in rabbit.

7.
China Pharmacy ; (12): 1203-1209, 2019.
Article in Chinese | WPRIM | ID: wpr-816964

ABSTRACT

OBJECTIVE: To establish the method for the rapidly non-destructive quality control of Liuwei dihuang capsule. METHODS: AOTF-NIR spectrometry was adopted. Taking 80 batches of Liuwei dihuang capsule produced by a manufacturer in recent three years as samples, HPLC chromatogram was adopted to determine the contents of loganin, morroniside, paeonol, paeoniflorin and ursolic acid; the content of water was determined according to general principles stated in 2015 edition of Chinese Pharmacopeia (part Ⅰ). Taking 70 batches of samples as correction set, the partial least square method and the cross-validation algorithm were used to establish the NIR quantitative model of 6 indexes in Liuwei dihuang capsules with the Unscrambler quantitative analysis software. Taking residual 10 batches of samples as validation set, external validation was conducted for the model. RESULTS: The correlation coefficients (R2) of internal and external validation of loganin, morroniside, paeonol, paeoniflorin, the content of water quantitative model were all greater than 0.9; the correction of standand deviation (RMSEC) were 0.372 8, 0.025 4, 0.263 3, 0.288 5, 0.186 7 and 0.037 7; the prediction of standard deviation (RMSEP) were 0.462 2, 0.077 5, 0.472 1, 0.634 9, 0.293 4 and 0.206 9; the external verification showed that mean deviations of preclicted value to actual value were 6.04%, 6.05%, 5.87%, 6.97%, 5.62% and 4.83%, with the mean deviation less than 10%.CONCLUSIONS:The established method can achieve rapidly non-destructive analysis Liuwei dihuang capsule.

8.
Article in Chinese | WPRIM | ID: wpr-707160

ABSTRACT

Objective To establish an HPLC method for the content determination of morroniside, sweroside, paeoniflorin and loganin of Liuwei Dihuang Decoction and its Cornus Officinalis-Cortex Moutan couple; To discuss the relationship between the whole prescription and the couple of main pharmacodynamic components. Methods The HPLC method was used at Hypersile C18 column (4.6 mm × 250 mm, 5 μm); the mobile phase consisted of methanol-water (24:76); the detective wavelength was 236 nm; the flow rate was 1.0 mL/min; the column temperature was 30 ℃. Results The linear ranges of morroniside, sweroside, paeoniflorin and loganin were among 0.480–7.680 μg (r=0.999 3), 0.103–1.650 μg (r=0.999 5), 0.120–1.920 μg (r=0.999 1) and 0.227–3.630 μg (r=0.999 7), respectively. The average recovery rates and RSD were 102.79%, 102.29%, 100.99%, 102.48%, and 1.73%, 1.48%, 1.32%, 0.75%, respectively. The contents of morroniside, sweroside and paeoniflorin in Liuwei Dihuang Decoction were slightly higher than that in Cornus Officinalis - Cortex Moutan couple, and the contents of loganin were almost the same. Conclusion The method is simple, stable, accurate and reproducible. It can be used for content determinate of glycosides in Liuwei Dihuang Decoction and Cornus Officinalis-Cortex Moutan couple. Cornus Officinalis-Cortex Moutan couple has the glycosides with tonifying kidney effect of Liuwei Dihuang Decoction.

9.
Article in Chinese | WPRIM | ID: wpr-851756

ABSTRACT

Objective To investigate the effects of morroniside extracted from root-bark of Sambucus williamsii on TNF-α-induced MC3T3-E1 cell inflammation, and explore its mechanism of action including Caspase, MAPK, and NF-κB signaling pathway. Methods MC3T3-E1 cells were cultured in medium containing TNF-α (50 ng/mL) and different doses of morroniside. The cell viability was examined by MTT assay, and the levels of IL-1β and IL-6 in the supernatants were identified by ELISA. The expressions of Caspase-3, Caspase-9, p-ERK, p-JNK, p-p38, p-IκBα, IκBα, and NF-κB on the protein level was tested by Western blotting. Results MTT assay results showed that morroniside could promote the proliferation of MC3T3-E1 cells and protect the MC3T3-E1 cells induced by TNF-α. ELISA results showed that the expressions of IL-1β and IL-6 were inhibited. Meanwhile, morroniside could reduce the expression of Caspase3, Caspase9, p-ERK, p-JNK, p-p38, and p-IκBα protein, and increase IκBα protein expression while there was no significant difference in the expression of NF-κB p65. Conclusion Morroniside has protective effect on TNF-α-induced MC3T3-E1 cells inflammation. The possible related mechanism is that morroniside inhibits the release of inflammatory cytokines, the activation of MAPK and NF-κB pathway, and the expression of Caspase, thereby inhibiting the apoptosis of MC3T3-E1 cells.

10.
Article in English | WPRIM | ID: wpr-727942

ABSTRACT

Our previous studies have confirmed that morroniside has neuroprotective effects. However, the effects of morroniside on cardiac myocardium remain unknown. Rats were anaesthetized with 10% chloral hydrate (0.35~0.4 mL/kg) and an acute myocardial infarction (AMI) was induced by ligating the anterior descending coronary artery (LAD). Following AMI, morroniside was administered intragastrically for 3 consecutive days at doses of 45, 90 and 180 mg/kg, respectively. Lactate dehydrogenase (LDH) and cardiac troponin T (cTnT) activities in AMI rats in the serum were detected with commercial kits. The expression of IL-6, IL-1β and TNF-α in myocardium was detected by Western blotting analysis. We observed a significant decline in the Q(q) wave amplitude in morroniside-treated rats after 72 h. Additionally, treatment of morroniside decreased the levels of LDH and cTnT in AMI rats. We also observed that morroniside reduced the expression of IL-6, IL-1β and TNF-α in myocardium. Taken together, our findings demonstrate that morroniside had effective anti-inflammatory properties in AMI rats.


Subject(s)
Animals , Blotting, Western , Chloral Hydrate , Coronary Vessels , Inflammation , Interleukin-6 , L-Lactate Dehydrogenase , Myocardial Infarction , Myocardium , Neuroprotective Agents , Rats , Troponin T
11.
Chinese Traditional Patent Medicine ; (12): 1845-1849, 2017.
Article in Chinese | WPRIM | ID: wpr-658750

ABSTRACT

AIM To establish a quantitative analysis of multi-components by single mark (QAMS) method for the simultaneous content determination of five constituents in Shanzhuyu Formulated Granules (Corni Fructus).METHODS The analysis of 80% methanol extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.3% phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 335 nm.With morroniside as an internal standard,the relative correction factors of gallic acid,5-hydroxymethylfurfural,loganin and cornuside were established,followed by the determination of their contents.RESULTS Gallic acid,5-hydroxymethylfurfural,morroniside,loganin and cornuside showed good linear relationships within the ranges of 0.0120-0.120,0.026 8-0.268,0.074 4-0.744,0.058 6-0.586 and 0.0086-0.086 μg (r≥ 0.9999),whose average recoveries (RSDs) were 103.43% (1.45%),103.36% (1.50%),104.47% (0.30%),102.08%(1.74%) and 104.01% (0.62%),respectively.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This stable and reliable method can be used for the quality control of Shanzhuyu Formulated Granules.

12.
Chinese Pharmaceutical Journal ; (24): 1212-1216, 2017.
Article in Chinese | WPRIM | ID: wpr-858637

ABSTRACT

OBJECTIVE: To study the chemical constituents from the ripe fruits of Cornus officinalis Sieb. et Zucc., and evaluate their protection on PC12 cells exposed to oxygen and glucose deprivation. METHODS: The compounds were isolated through various chromatographic methods including macroporous adsorption resin, silica gel, ODS, Sephadex LH-20 column chromatography, and preparative HPLC, and their structures were determined through spectroscopic analysis, including 1D and 2D NMR and MS data. RESULTS: Eight compounds were isolated from the water extract of the ripe fruits of Cornus officinalis Sieb. et Zucc., and their structures were elucidated as 6'-O-acetyl-7β-O-ethyl morroniside (1), chrysoderol(2), luteolin(3), 5-hydroxymethylfurfural(4), syringate(5),p-hydroxybenzoic acid(6), caffeic acid methyl ester(7), ethyl gallate(8). CONCLUSION: Compound 1 is a new iridoid glucoside, and compounds 2, 3, 5, 7 are obtained from Cornus officinalis for the first time. The MTT results show that compound 4 moderately increases the viability of OGD/R treated PC12 cells at the concentration of 1.0 μmol·L-1.

13.
Article in Chinese | WPRIM | ID: wpr-852943

ABSTRACT

Objective: To establish an HPLC fingerprint of Qinqi Rheumatism Formula (QRF) for its quality control and effective components determination. Methods: HPLC method was performed on Inert Sustain C18 (150 mm × 4.6 mm, 5 μm) column with gradient elution composed of acetonitrile-aqueous solution containing 0.1% phosphoric acid at the flow rate of 1.0 mL/min. The column temperature was set at 30℃, while the detective wavelength was set at 203 nm. The common mode of HPLC fingerprint for 10 batches of QRF was established with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2004 A edition) and the common peaks were identified by reference compounds. Results: Fingerprints of QRF were established. The similarities of the 10 batches of samples were above 0.99. A total of 19 common peaks were found. Eight mutual peaks were from Panacis Majoris Rhizoma, eight mutual peaks were from Gentianae Macrophyllae Radix, and five mutual peaks were from Corni Fructus (Peak 1 was the common peak from Panacis Majoris Rhizoma, Gentianae Macrophyllae Radix, and Corni Fructus). Based on the retention time of reference substances, eight constituents including loganic acid (peak 2), morroniside (peak 3), gentiopicroside (peak 5), loganin (peak 6), ginsenoside Ro (peak 12), ginsenoside F1 (peak 13), panax japonicus IVa (peak 14), and ginsenoside F2 (peak 17) were indentified. Conclusion: The method is stable, specific, and reproducible, and can be used for the quality control of QRF and the study of its effective components.

14.
Article in Chinese | WPRIM | ID: wpr-852905

ABSTRACT

Objective: To establish a method of quantitative analysis of multi-components by single marker (QAMS) for medicinal materials and pieces of Cornus officinalis. This method was used in combination with electronic-eye and electronic-tongue technique, and the best steaming time of Cornus officinalis was selected. Methods: Medicinal materials and pieces of C. officinalis were used as the research objects. The contents of five components were determined by establishing the relative correction factor (RCF) of gallic acid, 5-hydroxymethyl furfural (5-HMF), morroniside, cornuside, and internal reference loganin in C. officinalis. Color and taste were measured by electronic eye and electronic tongue technique. The data were analyzed by principal component analysis (PCA), and the best steaming time was optimized by analyzing the results of three methods. Results: The five compounds were well separated. The RSD values of precision and reproducibility were all less than 2%. The stability was good in 24 h. The linear relationship among the concentration and peak areas of the five compounds was all linear (r ≥ 0.999 6). The average recoveries were between 98% and 100.1% and the RSD values were all less than 2%; The RCFs of loganin with the other four compounds were 0.560, 1.344, 1.255, and 0.972 in a linear range. In the principal component analysis (PCA), the sums of main components were 94.618% and 94.98% and the discrimination indexes (DI) were 98 and 93, which indicated that all the samples of C. officinalis could be distinguished well by the electronic-eye and the electronic-tongue. The results showed that the optimum steaming time of C. officinalis was 4 h. Conclusion: The best steaming time of C. officinalis can be optimized by the combination of QAMS with electronic-eye and electronic-tongue techniques.

15.
Article in Chinese | WPRIM | ID: wpr-852681

ABSTRACT

Objective: To research endogenous metabolites changes of model group on human umbilical vein endothelial cells (HUVECs) injury induced by advanced glycation end products (AGEs) and the accommodation mechanism of morroniside (active components in Cornus officinalis) on the abnormal metabolism. To find potential biomarkers which morroniside protected the injured HUVECs, and to explore mechanisms of morroniside in treatment of HUVECs injury induced by AGEs. Methods: HUVECs were cultivated in vitro, HUVECs injury model was established by the induction of AGEs. Cells were divided into three groups, control group, model group, and treatment group. Cell samples of three groups were determined with UPLC-Q-TOF/MS. Pattern recognition methods including PCA, PLS-DA, and OPLS-DA were applied to analyze multivariate data, and t-test was used in significant statistical analysis. It was used to find out potential biomarkers. Metabolomic feature network graphs were constructed ingenuity pathway analysis (IPA). Results: In pattern recognition, control group, model group, and treatment group could be distinguished better from each other. According to VIP values, 30 ions which had a significant contribution to the classification were screened further, 10 potential metabolic biomarkers were identified qualitatively. These endogenous substances of the cells in treatment group in vivo had varying degrees of recovery. Five metabolic pathways were identified, and a metabolomic feature network of morroniside to protect against HUVECs injury induced by AGEs was constructed. Conclusion: Changed metabolities on HUVECs injury induced by AGEs can be certainly recovered by morroniside, and the treatment effect of morroniside can be connected with the regulation of five related metabolic pathways.

16.
Article in Chinese | WPRIM | ID: wpr-852588

ABSTRACT

Objective: To establish methods of qualitative identification and quantitative determination for Zishen Yangyin Granules (ZYG). Methods: The TLC method was used to identify the herb by mixture of chloroform-methanol (31) as a developing solvent on high performance silica gel precoated plate (HSGF254) and using 5% vanillic aldehyde sulfuric acid as a chromogenic reagent for qualitative identification of Corni Fructus; TLC identification of Eclipta prostrata, Alismatis Rhizoma, and Moutan Cortex was performed on high performance silica gel precoated plate (HSGF254) with petroleum ether (60-90 ℃)-chloroform-ethyl acetate (312) as developing solvent. The same developing method was used to identify E. prostrata, A. Rhizoma, and paneol in M. Cortex of ZYG by different detected method at the same time. The contents of morroniside, loganin, hyperoside, specnuezhenide, and paeonol were analyzed by high performance liquid chromatography on C18 column (250 mm × 4.6 mm, 5 μm) with the mobile phase of acetonitrile-0.1% trifluoroacetic acid by gradient elution. The detection wavelength was set at 254 nm. Results: Morroniside and loganin were used to identify C. Fructus in ZYG, paeonol in M. Cortex can be identified at 254 nm; Some substances in E. prostrata can be identified at 366 nm; Some substances in A. Rhizoma can be detected in sunlight, with 5% phosphomolybdic acid in ethanol as a chromogenic reagent. The TLC separation was desirable with moderate Rf value and clear spot. The methodology validation for the assay of morroniside, loganin, hyperoside, specnuezhenide, and paeonol presented that they were in good linear correlation in the ranges of 4.432-110.8, 4.192-104.8, 4.040-101.0, 4.132-103.3, and 4.076-101.9 μg/mL, The correlation coefficients of indicator were over 0.999 7. The average recoveries were between 96.57% and 98.67%. The RSD value of intra-day precision was less than 2% and the RSD value of inter-day precision was less than 3%. The method has good stability and reproducibility. Conclusion: The methods of quality control are specific, reproducible, accurate, and suitable, which can be successfully applied to the quality control of ZYG.

17.
Article in Chinese | WPRIM | ID: wpr-852384

ABSTRACT

Objective To establish an HPLC fingerprint of Xiaochuan Granula (XCG), and to make a quantitative analysis of seven components by fused-core column. Methods Kromasil C18 (150 mm × 4.6 mm, 3.5 μm) was used with the mobile phase of Methanol (A)-acetonitrile (B)-water (C), at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm for amygdalin and magnolin, 240 nm for morroniside, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol, schizandrin, and fingerprint; And the column temperature was maintained at 40 ℃. Common peaks had been identified by UPLC-Q-TOF/MS and standard compounds. Results The fingerprint chromatography included 20 mutual peaks, and the similarity was more than 0.90. Eleven chemical components were identified by UPLC-Q-TOF/MS and standard compounds, which were 3-morroniside, oxypaeoniflorin, 5-loganin, 6-prim-O-glucosylcimifugin, 9-4'-O-β-glucopyranosyl-5-O-methylvisamminol, 11-magnolin, 15-schisandrin, 16-schisandrol B, 18-schisantherin A, 19-deoxyschizandrin, and 20-γ-schizandrin B. Moreover, seven active components (morroniside, oxypaeoniflorin, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol,magnolin, and schisandrin) were quantified and the average recovery rates ranged from 97.3% to 103.8% with RSDs less than 2.0%; Seven components in 10 batches samples were morroniside 0.51-0.69 mg/g, amygdalin 5.01-5.95 mg/g, loganin 1.02-1.33 mg/g, prim-O-glucosylcimifugin 0.35-0.45 mg/g, 4'-O-β-glucopyranosyl-5-O-methylvisamminol 0.45-0.55 mg/g, magnolin 0.38-0.48 mg/g, schisandrin 0.89-1.08 mg/g, respectively, and RSD of each component was less than 11.0%. Conclusion The method for establishing HPLC fingerprint and quantitative analysis of seven components is rapid, simple, and accurate, and can be used for the quality control of XCG.

18.
Article in Chinese | WPRIM | ID: wpr-852317

ABSTRACT

Objective: To establish RP-HPLC strategy to simultaneously determine the terpenoids constitute of Corni Fructus preparation which is useful for the intervention of acute immunological liver injury in mice induced by concanavalin A (ConA), including morroniside, loganin, cornuside, oleanolic acid and ursolic, thus providing a scientific basis on the quality controls of Corni Fructus terpenoids and related medicinal preparations. Methods: RP-HPLC method and Zorbax SB-C18 column (150 mm × 4.6 mm, 5 μm) chromatographic column were employed; 2 mmol/L γ-cyclodextrin was added into the mobile phase containing acetonitrile and 0.1% ortho-phosphoric acid for gradient elution; The volume flow was 1.0 mL/min, the column temperature was 30 ℃ and the detection wavelengths were 240, 360, and 210 nm; The injection volume was 3 μL. Results: The linear range in morroniside, loganin, cornuside, oleanolic acid, and ursolic acid was 10.42—333.33, 23.44—750.00, 9.11—291.67, 10.42—333.33, and 13.02—416.67 mg/L, respectively; The average recovery in the selected samples was 95.60%—98.02%, RSD was 1.47%—1.89%; The repeatability RSD was 1.46%—1.71%; The stability RSD was 1.29%—1.76%; Six batches of the Corni Fructus terpenoids medicinal preparation contained the average quality of morroniside, loganin, cornuside, oleanolic acid, and ursolic acid respectively was 669.6—680.2, 850.1—869.5, 94.1—96.4, 164.3—166.1, and 85.6—87.6 mg/L. Conclusion: The method established in this study is a credible way to determine the concents of morroniside, loganin, ornuside, oleanolic acid, and ursolic acid in the Corni Fructus terpenoids medicinal preparation with its simplicity, good repeatability, high sensibility and recovery rate.

19.
Article in Chinese | WPRIM | ID: wpr-852314

ABSTRACT

Objective: To establish the HPLC fingerprint and determine nine components (geniposidic acid, morroniside, chlorogenic acid, geniposide, loganin, pinoresinol diglucoside, liquiritin, rutin, and glycyrrhizic acid) of Youguiyin, so as to provide a scientific basis for the quality control. Methods: HPLC analysis was performed on Pursuit XRs 5 C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 230 nm, and the column temperature was 30 ℃. Fingerprints of ten batches of Youguiyin were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed by comparing the retention time and UV spectra among 10 batches of Youguiyin. The formula ingredients and reference substance, and nine components content were determined. Results: The similarities of fingerprints of 10 batches of Youguiyin and reference fingerprints were all greater than 0.904. There were 30 mutual peaks marked in total, which were eight mutual peaks from Eucommia ulmoides, eight mutual peaks from Glycyrrhiza uralensis, six mutual peaks from Cornus officinalis, six mutual peaks from Aconitum carmichaeli, one mutual peaks from Cinnamomum cassia, and none of mutual peaks were originated from Dioscorea opposita and Lycium barbarum, and three of them cannot be originated. Based on the the identification of the common peaks, nine components [geniposidic acid (peaks 7), morroniside (peaks 9), chlorogenic acid (peaks 11), gardenoside (peaks 12), loganin (peaks 13), pinoresinol diglucoside (peaks 14), liquorice glycosides (peaks 18), rutin (peaks 21), and glycyrrhizic acid (peaks 30)] were identified and quantified. The quality faction of nine components were 87.6—119.1 μg/g, 323.6—365.6 μg/g, 108.3—124.1 μg/g, 79.5—85.0 μg/g, 171.7—188.0 μg/g, 163.0—238.3 μg/g, 64.5—53.3 μg/g, 159.8—168.5 μg/g, and 72.8—83.6 μg/g in raw material. Conclusion: The method established in this study is simple, accurate and highly reproducible, and can provide basis for quality control of Youguiyin.

20.
Chinese Traditional Patent Medicine ; (12): 1845-1849, 2017.
Article in Chinese | WPRIM | ID: wpr-661669

ABSTRACT

AIM To establish a quantitative analysis of multi-components by single mark (QAMS) method for the simultaneous content determination of five constituents in Shanzhuyu Formulated Granules (Corni Fructus).METHODS The analysis of 80% methanol extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.3% phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 335 nm.With morroniside as an internal standard,the relative correction factors of gallic acid,5-hydroxymethylfurfural,loganin and cornuside were established,followed by the determination of their contents.RESULTS Gallic acid,5-hydroxymethylfurfural,morroniside,loganin and cornuside showed good linear relationships within the ranges of 0.0120-0.120,0.026 8-0.268,0.074 4-0.744,0.058 6-0.586 and 0.0086-0.086 μg (r≥ 0.9999),whose average recoveries (RSDs) were 103.43% (1.45%),103.36% (1.50%),104.47% (0.30%),102.08%(1.74%) and 104.01% (0.62%),respectively.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This stable and reliable method can be used for the quality control of Shanzhuyu Formulated Granules.

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