Predictive Analysis of Quality Markers of Jiangxi Aurantii Fructus Based on Fingerprint and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo analyze and predict the potential quality markers （Q-Marker） in the Genuine medicinal materials Jiangxi Aurantii Fructus based on fingerprints and network pharmacology. MethodUltra-high performance liquid chromatography （UPLC） and gas chromatography-mass spectrometry （GC-MS） fingerprints were established for 18 batches of Jiangxi Aurantii Fructus ，combined with chemometric methods to screen out candidate Q-Marker components.Use network pharmacology to construct a "core component-target-pathway" network to predict the Q-Marker and core targets of Jiangxi Aurantii Fructus，and then verify the biological activity of Jiangxi Aurantii Fructus Q-Marker by molecular docking method. ResultThe 18 batches of Jiangxi Aurantii Fructus use UPLC，GC-MS fingerprints combined with chemometric analysis，a total of 9 Q-Marker candidate components were screened out.Through network pharmacological analysis，it is predicted that nobiletin，neohesperidin，meranzin，naringin and D-limonene are the Q-Marker of Jiangxi Aurantii Fructus，acting on the core targets transforming protein p21/H-Ras-1（HRAS），cellular tumor antigen p53 （TP53），mitogen-activated protein kinase 8 （MAPK8），transcription factor AP-1（JUN），glycogen synthase kinase-3 beta（GSK3B），tumor necrosis factor（TNF），cyclin-dependent kinase inhibitor 1（CDKN1A），cAMP-dependent protein kinase catalytic subunit alpha（PRKACA），cysteine aspartate-specific protease-9（Caspase-9），cyclic AMP-responsive element-binding protein 1（CREB1），exerting gastrointestinal motility and antidepressant，anti-inflammatory，anti-tumor，etc.； molecular docking shows that nobiletin，neohesperidin，meranzin，naringin and D-limonene and the selected 10 core targets have good binding ability，reflecting the better biological activity of the Q-Marker of Jiangxi Aurantii Fructus. ConclusionThe Q-Marker of Jiangxi Aurantii Fructus can be comprehensively predicted from the two aspects of volatile and non-volatile components，providing a reference for the quality control of Jiangxi Aurantii Fructus and the further study of its pharmacodynamic mechanism.

Simultaneous Determination of 5 Components in the Classical Formula Huaihua San by HPLC Wavelength Switching Method / 中国药房

OBJECTIVE：To develop a method for simultaneous determination of 5 components in classical formula Huaihua san，including rutin ，naringin，neohesperidin，quercetin and pulegone. METHODS ：HPLC wavelength switching method was adopted. The determination was performed on Cosmosil C 18 column with mobile phase consisted of acetonitrile- 0.05％ phosphoric acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 257 nm for rutin ，283 nm for naringin and neohesperidin ，254 nm for quercetin ，252 nm for pulegone ，respectively. The column temperature was set at 30 ℃， and sample size was 10 μL. RESULTS：The linear range was 21.7-2 170 μg/mL for rutin，46-4 600 μg/mL for naringin，22.3- 2 230 μg/mL for neohesperidin，0.96-96 μg/mL for quercetin，2.7-270 μg/mL for pulegone（all r＞0.999），respectively. RSDs of precision，stability（24 h）and reproducibility tests were all lower than 2％（n＝6）. Average recoveries were 100.70％，99.31％， 101.10％，100.03％ and 99.63％（all RSD ＜2％，n＝9）. Among 3 batches of Huaihua san samples ，the contents of above 5 components were 20.055-22.615，25.557-27.806，11.428-13.250，0.350-0.478，2.372-4.011 mg/g，respectively. CONCLUSIONS ： Established method is simple ，accurate and reproducible ，and could be used for the simultaneous determination of 5 components in Huaihua san.

Therapeutic effect of neohesperidin on TNF-α-stimulated human rheumatoid arthritis fibroblast-like synoviocytes / 中国天然药物

During the pathogensis of rheumatoid arthritis (RA), activated RA fibroblast-like synoviocytes (RA-FLSs) combines similar proliferative features as tumor and inflammatory features as osteoarthritis, which eventually leads to joint erosion. Therefore, it is imperative to research and develop new compounds, which can effectively inhibit abnormal activation of RA-FLSs and retard RA progression. Neohesperidin (Neo) is a major active component of flavonoid compounds with anti-inflammation and anti-oxidant properties. In this study, the anti-inflammation, anti-migration, anti-invasion, anti-oxidant and apoptosis-induced effects of Neo on RA-FLSs were explored to investigate the underlying mechanism. The results suggested that Neo decreased the levels of interleukin IL-1β, IL-6, IL-8, TNF-α, MMP-3, MMP-9 and MMP-13 in FLSs. Moreover, Neo blocked the activation of the MAPK signaling pathway. Furthermore, treatment with Neo induced the apoptosis of FLSs, and inhibited the migration of FLSs. It was also found that Neo reduced the accumulation of reactive oxygen species (ROS) induced by TNF-α. Taken together, our results highlighted that Neo may act as a potential and promising therapeutic drug for the management of RA.

Simultaneous Determination of the Contents of 12 Flavonoids in Quzhiqiao from Different Collection Places by HPLC / 中国药房

OBJECTIVE：To establish a method for the simultan eous determination of the contents of 12 flavonoids in Quzhiqiao. METHODS ：HPLC method was adopted. The determination was performed on Agilent Extend C 18 column with mobile phase consisted of 0.1％ formic acid-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃. The detection wavelength was set at 330 nm，and sample size was 10 μL. The contents of 12 components（such as eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone ）in 10 batches of Quzhiqiao from different collection places were determined. RESULTS ：The linear range of eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone were 1.65-16.51，4.50-45.02，35.41-354.12，4.11-41.12，2.29-22.86，34.96-349.56，1.42-14.15，1.50-15.04， 1.83-18.28，1.51-15.08，1.61-16.12，1.28-12.84 μg/mL，respectively（all r＞0.999 7）. The detection limits were 0.165 1，0.450 2， 3.541 2，0.411 2，0.228 6，3.495 6，0.141 5，0.150 4，0.182 8，0.150 8，0.161 2，0.128 4 μg/mL，respectively. The limits of quantitation were 0.547 8，1.487 4，11.663 3，1.360 3，0.758 3，11.594 9，0.466 3，0.497 1，0.601 2，0.499 9，0.532 3，0.424 6 μg/mL，respectively. RSDs of precision （n＝6），reproducibility（n＝6）and stability （24 h，n＝7）tests were all lower than 3％. The average recoveries were 99.50％，99.61％，98.18％，98.85％，98.48％，98.50％，98.25％，99.91％，103.13％，98.82％， 98.44％ ， 100.29％ （RSD＝1.49％ -2.38％ ， n＝6）. The contents of the above 12 components in 10 batches of samples from different collection places were 1.995 5-2.648 8，4.317 7- 5.005 1，33.215 5-34.054 6，3.140 4-3.471 5，3.221 2-3.748 8， 42.746 6-44.026 6，0.202 7-0.239 4，0.191 2-0.208 8，0.080 3- 0.097 9，0.291 9-0.307 1，0.119 9-0.149 1，0.082 7-0.089 8 mg/g. CONC LUSIONS：The method is accurate ，reliable，simple and efficient，which can be used to simultaneous determination of the contents of 12 flavonoids in Quzhiqiao ，and to provide reference for the establishment of quality control standards of Quzhiqiao.

Investigation of Chemical Constituents and Sources of Zhishi Xiebai Guizhi Decoction by LC-MS/MS / 中国药学杂志

OBJECTIVE: To study the chemical constituents of Zhishi Xiebai Guizhi decoction and its single agent by high performance liquid chromatography-mass spectrometry to provide a scientific basis for the quality evaluation and control. METHODS: Agilent XDB-C18(4.6 mm×250 mm, 5 μm) column was used for HPLC analysis with acetonitrile-water as mobile phase by gradient elution. Using liquid chromatography-mass spectrometry technology to collect data in positive and negative ion mode, combined with literature reports, the main chemical components of Zhishi Xiebai Guizhi decoction were analyzed and identified. RESULTS: A total of 19 compounds were identified from Zhishi Xiebai Guizhi decoction. The main components are naringin, neohesperidin, etc. The main component types include alkaloids, steroidal saponins, glycosides, flavonoids, etc. CONCLUSION: This study comprehensively clarified the chemical composition of Zhishi Xiebai Guizhi decoction, and it has important reference value for the identification and quality evaluation of Zhishi Xiebai Guizhi decoction.

Metabolic characteristics of active parts of lipid-lowering flavonoid extract of Daidai in liver and intestinal microsomes of rats / 中国中药杂志

The paper studies and compares the metabolic difference of active ingredients of lipid-lowering flavonoid extract of Daidai in rat livers and intestinal microsomes,in order to explore the phase Ⅰ metabolism characteristics of active ingredients in livers and intestines. UPLC-MS/MS was used to establish a quantitative analysis method for active ingredients,neohesperidin and narngin,in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes. Differential centrifugation was used to make liver and intestinal microsomes of rats. A phase Ⅰ metabolism incubation system was established,and the concentrations of the residual at different incubation time points were analyzed. Graphs were plotted to calculate the metabolic elimination half-life of the main active parts,with the natural logarithm residual percentage values ln( X) at different time points as the y axis,and time t as the x axis. The metabolism characteristics of the active ingredients were compared. The established UPLC-MS/MS quantitative analysis method has a good specialization,standard curve and linear range,accuracy and precision,with a satisfactory lower quantitative limit. The method allows quantitative detection of the active ingredients in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes of rats. In the rats liver microsomes incubation system,the metabolic elimination half-life of neohesperidin and narngin were( 2. 20 ± 0. 28) h and( 1. 97±0. 28) h respectively. The elimination half-life of neohesperidin was larger than that of narngin,but with no statistically significant difference. In the rats intestinal microsomes incubation system,the metabolic elimination half-lives of neohesperidin and narngin were( 3. 68±0. 54) h and( 2. 26±0. 13) h respectively. The elimination half-life of neohesperidin was larger than that of narngin,with statistically significant differences( P<0. 05). The elimination half-lives of the active ingredients in liver microsomes were smaller than those in intestinal microsomes. The experiment results showed that the active ingredients of lipid-lowering flavonoid extract of Daidai had different elimination half-lives in phase Ⅰ rats liver and intestinal microsomes incubation system. This implied that they had different metabolic characteristics in rats liver and intestine,and liver may be the main metabolism site of the active ingredients. The phaseⅠ metabolism of narngin was stronger than that of neohesperidin. The differences between their metabolic characteristics may be related to the binding sites of B-ring hydroxyl in flavonoid glycosides and the number of methoxyl group. The results provided an important experimental basis for further development and clinical application of lipid-lowering flavonoid extract preparation of Daidai.

HPLC Determination of Four Flavonoids of Aurantii Fructus Immaturus in Niuhuang Qingwei Pills and Genetic Analysis of Medicinal Materials / 中国实验方剂学杂志

Objective: To establish an high performance liquid chromatography (HPLC) method for the simultaneous determination of four constituents in Niuhuang Qingwei pill (narirutin,naringin,hesperidin,and neohesperidin), and identify the source of Fructus Aurantii Immaturus. Method: The analysis was performed on a Waters CORTECS C18 column (4.6 mm×50 mm,2.7 μm), with acetonitrile-0.12% formic acid solution as the mobile phase for gradient elution. The flow rate was 0.5 mL·min-1, the detection wavelength was set at 283 nm, and the column temperature was 27℃. Result: 12 batches of Niuhuang Qingwei pills showed the different content of flavonoids as Citrus aurantium and C. sinensis. Narirutin,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.47-2 735 ng (r=0.999 6),7.25-3 625 ng (r=0.999 5),8.41-4 205 ng (r=0.999 4) and 8.36-4 180 ng (r=0.999 5),and their average recoveries were 101.3% (n=6,RSD 2.9%),98.0% (n=6,RSD 1.8%),95.9% (n=6,RSD 0.8%) and 96.0% (n=6,RSD 1.1%), respectively. The contents of narirutin,naringin,hesperidin,neohesperidin and total flavonoids were 0.36-1.28,2.66-4.87,1.02-11.07,3.58-6.41,and 7.98-13.34 mg·g-1, respectively. Conclusion: The developed method was simple,accurate and reliable,which can be used to identify the source of Aurantii Immaturus Fructus and simultaneously determine the content of four flavonoids in Niuhuang Qingwei pills. It could provide basic research for quality control and composition comparison of 2 kinds of Niuhuang Qingwei pills, showing more comprehensive indicators and reference value for the quality standard improvement of Niuhuang Qingwei pill.

Comparative pharmacokinetics of naringin and neohesperidin after oral administration of flavonoid glycosides from Aurantii Fructus Immaturus in normal and gastrointestinal motility disorders mice / 中草药·英文版

Objective: To compare the pharmacokinetics of naringin and neohesperidin after oral administration of Zhishi total flavonoid glycosides (ZSTFG) extracted from Aurantii Fructus Immaturus in normal and gastrointestinal motility disorders (GMD) mice. Methods: ZSTFG was orally given to normal and GMD mice induced by atropine or dopamine. The plasma samples were incubated with β-glucuronidase/sulfatase, the total (free + conjugated) naringenin and hesperitin were extracted with acetonitrile. The validated HPLC-MS/MS method was successfully applied to the pharmacokinetic study. Results: The results showed that, compared with the normal group, AUC0–∞, AUC0–t and Cmax for total naringenin and hesperitin were significantly higher (P < 0.01 or P < 0.05), while CLZ/F for total naringenin and hesperitin was significantly lower (P < 0.01) in the GMD group. Tmax, t1/2z, MRT0-t, and MRT0-∞ for naringenin were longer (P < 0.01) in the GMD group than those in the normal group. Conclusion: The results showed that there were significant differences in pharmacokinetic parameters of naringenin and hesperitin between normal and GMD groups. It was suggested that the absorption of naringenin and hesperitin was increased, and the elimination processes of naringenin and hesperitin were slower in the GMD group than the normal group. The data are of value for further pharmacological studies of ZSTFG and would be useful to provide a reference for improving the therapeutic regimen of ZSTFG in clinical trials.

Binding characteristics of plasma protein in active parts of Daidai lipid-lowering flavonoid extract / 中国中药杂志

To study the binding capacity of active ingredients of Daidai lipid-lowering flavonoid extract and plasma protein,investigate the ways to improve the traditional formula for calculating protein binding rates based on ultrafiltration,and increase the stability and reliability of the experimental results. UPLC-MS/MS was used to establish a quantitative analysis method for simultaneous determination of active ingredients( neohesperidin and narngin) in ultrafiltrate. The protein binding rates were calculated by the traditional ultrafiltration formula. The correction factors( F) were introduced later,and the binding rates calculated with the correction factors were compared with those without the correction factors. The binding capacity of the extract and plasma protein was evaluated. The quantitative analysis method established by UPLC-MS/MS had a good specificity. The standard curve and linear range,method accuracy,precision and lower limit of quantitation all met the requirements. The method met the requirement for quantitative detection of the active ingredients in ultrafiltrate after the rat plasma was filtrated in the ultrafiltration tube. Under the experimental conditions,the binding rates of both active ingredients( neohesperidin and narngin) were higher than 90%. The active ingredients and rat plasma protein were bound in a concentration-dependent manner,with statistically significant differences( P<0. 01). There was no statistically significant difference between the protein binding abilities of the two active ingredients with rat plasma protein. Therefore,the active ingredients of Daidai lipid-lowering flavonoid extract had a relatively strong binding strength with rat plasma protein,and they were bound in a concentration-dependent manner. Additionally,when calculating protein binding rates by the traditional ultrafiltration formula,the correction factors could be introduced to effectively reflect the errors of multiple ingredient groups in traditional Chinese medicine extracts.This correction method could provide a reference thinking and practical reference for the improvement of the determination method of the traditional Chinese medicine plasma protein binding ability based on ultrafiltration.

Effect of 60Co Irradiation on 3 Active Ingredients in Compound Hemostatic Capsules / 中国药师

Objective: To establish an HPLC method for the simultaneous determination of 3 active components in compound he-mostatic capsules and investigate the content changes before and after 60Co-γ ray irradiation. Methods: An Agilent-C18column (250 mm×4. 6 mm, 5 μm) was adopted and the flow rate was 1. 0 ml·min-1. The mobile phase consisted of acetonitrile-0. 5% phosphor-ic acid solution (triethylamine was used to adjust pH to 6) with gradient elution, the detection wavelength was 254 nm (0-35 min) and 281 nm (35-55 min), and the column temperature was 35℃. The compound hemostatic capsules were irradiated at 5, 8 and 10 kGy, respectively, and the contents of the 3 active components in compound hemostatic capsules were compared before and after the radia-tion,and the t-test was applied to investigate the significance. Results: Typhaneoside,isorhamnetin-3-O-neohesperidin and tetrahydro-palmatine respectively within the concentration range of 0. 018-0. 11 mg·ml-1,0. 020-0. 120 mg·ml-1and 0. 011-0. 066 mg·ml-1 showed a good linear relationship with the peak area. The average recovery was 97. 7% , 98. 7% and 99. 3% with the RSD of 0. 9% , 0. 9% and 0. 5% , respectively (n=6). After the irradiation, the contents of the three active components in compound hemostatic capsules all showed changes. Under the dose of 8 kGy, the content changes of typhaneoside and isorhamnetin-3-O- neohesperidin showed no statistical significance (P>0. 05),and when the dose increased to 10 kGy,the content of tetrahydropalmatine exhibited sig-nificant difference (P<0. 05). Conclusion: The established determination method for compound hemostatic capsule shows such ad-vantages as high recovery, good repeatability, simple operation and promising separation, which can be used as a quality control meth-od for compound hemostatic capsules. The sterilization effect of 60Co-γ ray irradiation on compound hemostatic capsules is significant without notable changes in the active components. When the irradiation dose is equal to or below 5 kGy,the change of each component is not obvious,which can provide reference for the radiation sterilization of compound hemostatic capsules.

Comparison of content of five flavones in Citrus grandis “Tomentosa” and Citrus grandis with quantitative analysis of muli-components by single marker / 中草药

Objective To establish quantitative analysis of multi-components by single marker (QAMS) to compare five flavones in Citrus grandis “Tomentosa” and C. grandis. Methods Four relative correction factors (RCFs) of neohesperidin, rhoifolin, naringenin, apigenin were established in the HPLC method with the Naringin as internal standard, which were used to determine the content of four flavones in C. grandis “Tomentosa” and C. grandis. Meanwhile, external standard method (ESM) was employed to calculate the content of five flavones. The difference between ESM and QAMS were analyzed to evaluate the accuracy of QAMS. T-test was used to compare five flavones in C. grandis ‘Tomentosa’ and C. grandis. Results RCFs of neohesperidin, rhoifolin, naringenin, apigenin were 0.898, 1.519, 0.313, 0.406, and repeatability was good in different experimental conditions (RSD < 3.0 %). There were no significant differences in the quantitative analysis results of two methods. Content of naringin, rhoifolin and naringenin in two species were significant different. Conclusion The RCFs of neohesperidin, rhoifolin, naringenin, apigenin as reference to naringin are accurate and feasible. Content of flavones in Citrus grandis “Tomentosa” and C. grandis are obvious different. QASM can be applied as a strategy for quality control of C. grandis.

Simultaneous quantitative determination of nine active compounds from Aurantii Fructus by HPLC / 中草药

Objective: To establish an HPLC method for simultaneous determination of 7-hydroxycoumarin, narirutin, naringin, hesperidin, neohesperidin, nobiletin, 3,5,6,7,8,(3,4)-heptamethoxyflavone, tangeretin, and auraptene from Aurantii Fructus, determinate and compare the content of various chemical components from Aurantii Fructus with different perimeters. Methods: The HPLC system consisted of a diamon C18 (250 mm × 4.6 mm, 5 μm), the mobile phase was methanol (A)-water (adjusted pH 3.0 with phosphorous acid). The gradient elution conditions: 0-25 min, 30%-50% A; 25-35 min, 50%-70% A; 35-40 min, 70%-75% A; 40-55 min, 75%-100% A. The flow rate was 1.0 mL/min and the column temperature was 30 ℃. The detection wavelength was 320 nm and the injection volume was 20 μL. Results: The calibration curves of 7-hydroxycoumarin, narirutin, naringin, hesperidin, neohesperidin, 3,5,6,7,8,(3,4)-heptamethoxyflavone, nobiletin, tangeretin, and auraptene had good linear relationship in the ranges of 0.002 18-0.07 μg (r = 0.999 8), 0.021 8-0.70 μg (r = 0.999 9), 0.242 5-7.76 μg (r = 0.999 8), 0.024 38-0.78 μg (r = 0.999 4), 0.523 76-16.76 μg (r = 0.999 3), 0.003 13-0.10 μg (r = 0.999 3), 0.004 13-0.132 μg (r = 0.999 6), 0.002 75-0.088 μg (r = 0.999 6), and 0.000 93-0.03 μg (r = 0.999 3); And the average recoveries (n = 6) of the nine components were 98.50%, 98.80%, 99.51%, 98.43%, 99.64%, 99.21%, 100.03%, 98.75%, and 101.11%, respectively. Conclusion: This method can be applied to determinating the components from Aurantii Fructus, including 7-hydroxycoumarin and 3,5,6,7,8,(3,4)-heptamethoxyflavone, etc.

Optimization of matrix formulation for Shaofu Zhuyu Cataplasm and research on its transdermal permeability in vitro / 中草药

Objective To optimize the matrix formulation for Shaofu Zhuyu Cataplasm (SZC) by Plackett-Burman design combined with Box-Behnken response surface method and study its transdermal permeation properties in vitro. Methods In this paper, the appearance description, initial bonding strength, endurance bonding strength, and peel strength were taken as indexes. Based on the result of a single factor experiment, the formula for the SZC was optimized by Plackett-Burman combined with Box-Behnken method. Franz diffusing cells method was chosen to investigate the tansdermal infiltration capacity of SZC with different dosage penetration enhancers of azone in vitro, taking ferulic acid, paeoniflorin, isorhamnetin-3-O-neohesperidin, and typhaneoside as index components. Results NP-700, tartaric acid and kaolin had significant effects on the properties of SZC. The optimal ratio of the prescription was as follows: NP-700-tartaric acid-kaolin (2.42:0.16:0.96). The promoting effect of 3% concentration for index component was better, and the transdermal rates of ferulic acid, paeoniflorin, isorhamnetin-3-O-neohesperidin, and typhaneoside were 6.889 4, 1.917 4, 0.852 0, and 0.893 7 μg/(cm2∙h), respectively. Conclusion The SZC was uniform, without residual behavior in application. And it had a good release and transdermal properties, and transdermal actions were consistent with zero-order kinetics.

Quality Evaluation of Shuwei Mixture Based on Multi-index Constituents / 医药导报

Objective The quality evaluation of the Shuwei mixture was determined by the content of the six components of sinapine cyanide sulfonate, magnolol, honokiol, hesperidin, naringin and neohesperidin. Methods RP-HPLC method was used.The separation was performed on a Agilent ZORBAX Eclispse SB-C18column (4.6 mm×250 mm,5 mm),the mobile phase consisted of acetonitrile(A)-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL·min-1.The detection wavelength was 326 nm ( sinapine cyanide sulfonate ), 294 nm ( magnolol, honokiol ) and 283 nm ( naringin, neohesperidin).The column temperature was kept at 30 ℃. Results The sinapine cyanide sulfonate, magnolol, honokiol, hesperidin,naringin and neohesperidin all had good linear relationship in the ranges of 0.049 6-1.24,0.048 2-1.205,0.060 5-1.512 5,0.187 2-4.68,0.131 6-3.29,0.197-4.925 μg.The average recoveries were 100.66%, 99.86%, 101.37%, 102.41%, 99.01%, 102.05%, respectively, RSD were 0.82%, 1.89%, 2.56 %, 0.74%, 1.54%, 0.99%, respectively. Conclusion The method is simple,accurate,reproducible and nice to the separation,and can be used for the quality evaluation of Shuwei mixture.

Simultaneous determination of five constituents in Simotang Oral Liquid by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of five constituents in Simotang Oral Liquid (Aucklandiae Radix,Aurantii Fructus,Arecae Semen,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent Zorbax C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 283 nm.RESULTS Synephrine,norisoboldine,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.8-185.6 μg/mL (r =0.999 9),0.829-26.52 μg/mL (r =1.000 0),9.775-312.8 μg/mL (r =1.000 0),0.594-19 μg/mL (r =0.999 5) and 5.2-166.4 μg/mL (r =1.000 0),whose average recoveries were 98.93％,98.95％,102.57％,99.67％ and 103.43％ with the RSDs of 1.85％,1.27％,0.52％,0.89％ and 0.43％,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the rapid quality control of Simotang Oral Liquid.

Simultaneous Determination of Naringin and Neohesperidin in Weitong Pills by HPLC / 中国药师

Objective: To establish a method for the content determination of naringin and neohesperidin in Weitong pills.Methods: An HPLC method was adopted.The determination was performed on an Agilent TC-C18 column with the mobile phase consisting of acetonitrile-water(20∶80)at the flow rate of 1.0 ml·min-1.The detection wavelength was set at 283nm.The column temperature was 30 ℃.Results: Naringin and neohesperidin respectively within the concentration range of 0.027-4.552 μg (r=0.999 9) and 0.029-4.016 μg(r=0.999 8) had good linear relationship with the peak area, the average recovery was 102.19% and 103.60%,and the RSD was 2.88% and 1.79%(n=9), respectively. Conclusion: The method is simple,accurate and reproducible, and suitable for the content determination of naringin and neohesperidin in Weitong pills.

Pharmacokinetic study on compatibility of berberine and citrus aurantium extract in rats / 药物评价研究

Objective UPLC-MS/MS bio-analysis method was developed for the simultaneous determination ofberberine,naringin,hesperidin,and neohesperidin in plasma of rats.Methods UPLC Acquity BEH C18 (50 rmm × 2.1 mm,1.7 μm) column was used,mobile phases were containing 0.05％ formic acid and 2 mmol/L ammonium formate in water (A)-containing 0.05％ formic acid in acetonitrile (B) as the mobile phase gradient elution;SD rats were randomly divided into oral administration berberine group,Citrus aurantium extract group,and berberine and C.aurantium extract compatibility group.Results UPLC-MS/MS method could be applied to determination of berberine,naringin,hesperidin,and neohesperidin,method validation meets the requirements of biological sample analysis.When rats were administered with berberine and C.aurantium extract compatibility,the plasma concentration of berberine was much more than single dose of berberine group and the bioavailability of berberine was increased.Meanwhile,naringin and neohesperidin can be detected in rat's plasma.Conclusion The bioavailability of flavonoids is significantly improved as well compared to the single dose of C.aurantium extract.This suggests that berberine and C.aurantium extract compatibility has significant drug-drug interaction.

Study on the Mechanism of Aurantii Fructus Immaturus and Its Main Active Ingredients in Promoting Gastrointestinal Motility of Model Rats with Spleen Deficiency / 中国药房

OBJECTIVE:To study the mechanism of Aurantii fructus immaturus(AFI)and its main active ingredients in pro-moting gastrointestinal motility of model rats with spleen deficiency. METHODS:170 rats were randomly divided into blank group (10 rats) and modeling group (160 rats),rats in modeling group was induced models with spleen deficiency by bitter cold diar-rhea+irregular diet. After modeling, rats were randomly divided into model group, naringin (NA) low-dose, medium-lose, high-dose groups(3.267,6.535,13.070 mg/mL),neohesperidin(NE)low-dose,medium-lose,high-dose groups(3.865,7.730, 15.460 mg/mL),synephrine(SY)low-dose,medium-lose,high-dose groups(0.252,0.504,1.008 mg/mL),compatibility groups with 3 monomer ingredients (NA-NE-SY) low-dose,medium-lose,high-dose and AFI water decoction low-dose,medium-lose, high-dose groups(0.104,0.208,0.416 g/mL,calculated by crude drug),ig,once a day,10 mL/kg,for 7 d. After the last admin-istration,gastrin (GAS) in serum,and acetylcholine (ACh),motilin (MTL),substance P (SP),vasoactive intestinal peptide (VIP)levels in plasma were detected. RESULTS:Compared with blank group,GAS level in serum and ACh,MTL,SP levels in plasma in model group were reduced(P<0.01),VIP level in plasma was increased(P<0.05). Compared with model group,ex-cept for the GAS level in serum showed no obvious change in NA high-dose group and SY doses groups,other medicine groups were obviously increased (P<0.05 or P<0.01);the ACh levels in serum were obviouly increased in NE high-dose group,SY high-dose group and AFI water decoction low-dose group(P<0.01). MTL levels in plasma were obviously increased in NE medi-um-dose,high-dose groups,SY high-dose group,compatibility low-dose,medium-dose groups and AFI water decoction medi-um-dose,high-lose groups (P<0.05);SP levels in plasma were obviously increased in NA low-dose,medium-dose groups and NE doses groups(P<0.05 or P<0.01);VIP levels were reduced in NA low-dose group,SY high-dose group and AFI water decoc-tion low-dose,medium-lose groups(P<0.05). CONCLUSIONS:AFI may promote the gastrointestinal motility of model rats with spleen deficiency by promoting the secretion of GAS,ACh,MTL,and inhibiting the secretion of VIP;there are differences be-tween AFI and the 3 monomer ingredients in regulation of gastrointestinal hormones.

Simultaneous Determinations of Four Ingredients in Yiqi Hewei Capsules by HPLC / 医药导报

Objective To establish a method for the quantitative determinations of the ingredients in yiqi hewei capsules. Methods RP-HPLC method was used. The separation was performed on a Thermo C18 column with mobile phase consisting of acetonitrile-water (20: 80) (pH was adjusted to 3 with glacial acetic acid) at the flow rate of 1. 0 mL·min-1. The detection wavelength was 283 nm. Results The linear ranges were ( 0. 5735 -5. 7360) × 10 -2 mg·mL-1 for baicalin, (0. 1940-1. 9395) × 10 -2 mg·mL-1 for hesperidin, (0. 2051 -2. 0510) × 10 -3 mg·mL-1 for naringin and (0. 2020 -2.0200) ×10 -2 mg·mL-1 for neohesperidin;RSD =0.28%,0.54%,0.54%,0.31%(n =5),respectively. The average recoveries were 103. 94%, 98. 06%, 103. 09%, 95. 67%, respectively. Conclusion The established method is simple, sensitive and accurate,which can be used for the quality control of yiqi hewei capsules.

Simultaneous determination of four constituents in Shugan Pills by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Shugan Pills (Toosendan Fructus,Corydalis Rhizoma,Paeoniae Radix Alba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 35 ℃ thermostatic C18 column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.05％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 230 nm.RESULTS Paeoniflorin,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 0.040 44-1.011,0.045 20-1.130,0.047 08-1.177 and 0.047 24-1.181 μg (r =0.999 9),whose average recoveries were 100.5％,98.6％,98.4％ and 101.3％ with the RSDs of 1.5％,0.9％,1.2％ and 1.1％,respectively.The contents of various constituents in the samples with three dosage forms from seven manufacturers showed obvious differences.CONCLUSION We should pay attention to the uneven quality of Shugan Pills.