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OBJECTIVE: To investigate the absorption mechanism of paeoniflorin in Radix Paeoniae Alba, both of Radix Paeoniae Alba and Radix Angelica Sinensis and Siwu Decoction. METHODS: The circulatory perfusion technique was used in this study, the concentrations of paeoniflorin and phenol red in intestinal absorption circulation were respectively determined by HPLC and UV. The effects of pH value, drug concentration, absorption site and P-gp on the absorption of paeoniflorin were investigated separately. RESULTS: By comparing the absorption of paeoniflorin in Radix Paeoniae Alba, both of Radix Paeoniae Alba and Radix Angelica Sinensis, and Siwu Decoction, it was found that in the sample solution at the same absorption site, pH value and concentration (based on the concentration of paeoniflorin), the absorption, Ka and cumulative absorption of paeoniflorin in compound Siwu decoction group were significantly increased compared with the other groups (P<0.05), while t1/2 was significantly decreased (P<0.05). When combined with the inhibitor and inducer of P-gp, the absorption of paeoniflorin showed significant increase or decrease in the amount of absorption, Ka and cumulative absorption% compared with the control group (P<0.05), and t1/2 also showed significant decrease or increase (P<0.05). CONCLUSION: The absorption of paeoniflorin could be affected by P-gp, and under the same conditions, the absorption of paeoniflorin in complicated Chinese herbal formula is better than that in the single herb and herb-pair.
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OBJECTIVE： To establish a method for simultaneous determination of matrine， oxymatrine， gallic acid， peoniflorin， costunolide and dehydrocostus lactone in Libiling tablets. METHODS： RP-HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelengths were 210 nm （matrine， oxymatrine） and 225 nm （gallic acid， peoniflorin， costunolide， dehydrocostus lactone）. The column temperature was set at 30 ℃， and sample size was 5 μL. RESULTS： The linear ranges of matrine， oxymatrine， gallic acid， peoniflorin， costunolide， dehydrocostus lactone were 0.053-5.28 mg/mL（r＝0.999 8）， 0.125-12.54 mg/mL（r＝0.999 9）， 0.013-1.33 mg/mL（r＝0.999 8）， 0.169-16.94 mg/mL（r＝0.999 9）， 0.048-4.77 mg/mL（r＝0.999 8）， 0.072-7.16 mg/mL （r＝0.999 9）. The limits of quantitation were 4.08×10－4， 4.48×10－4， 3.12×10－4， 2.10×10－4， 1.36×10－4， 1.84×10－4 mg/mL， respectively. The limits of detection were 1.24×10－4， 1.50×10－4， 1.02×10－4， 6.20×10－5， 4.20×10－5， 6.40×10－5 mg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2％ （n＝6）. The recoveries were 98.03％-101.43％ （RSD＝1.25％， n＝6）， 97.73％-102.26％ （RSD＝1.96％， n＝6）， 97.18％-101.41％ （RSD＝1.98％，n＝6）， 97.45％-102.11％ （RSD＝1.88％，n＝6）， 96.85％-101.07％ （RSD＝1.75％， n＝6）， 97.12％-102.64％ （RSD＝1.82％，n＝6）， respectively. CONCLUSIONS： Established method is simple， stable and rapid， and can be used for simultaneous determination of 6 components in Libiling tablets.
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Paeoniflorin is the main effective component of Paeoniae lactiflora and Paeonia suffruticosa. A large number of studies have proven that paeoniflorin has many pharmacological effects, such as anti-depression, anti-inflammation, analgesia, anti-tumor, liver protection, nerve protection, sedation and hypnosis, immunomodulation and so on. It has little toxic and side effects and has been highly concerned by people. At present, paeoniflorin is rarely used in clinical practice in the form of monomers. This article mainly refers to the related research literatures on paeoniflorin pharmacological action in recent three years, combs and summarizes the new progress of its pharmacological action research. The relationship between the drug effect and the prescription of traditional Chinese medicine was discussed, in order to provide a reference for further development and clinical application of paeoniflorin.
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Objective To establish the quality standard of Wuji-Jianwei granules. Methods The contents of Bupleuri Radix, Codonopsis Radix, Paeoniae Radix Alba, Corydalis Rhizoma in Wuji-Jianwei granules were identified by using the thin layer chromatography (TLC) method. The contents of peoniflorin was determined by the high performance liquid chromatography (HPLC) method. Results The TLC had strong specificity and high separation, negative control without interference. The linear range was 0.124 8-0.748 8 μg for peoniflorin (r=0.999 6). The average recovery was 94.42% (RSD=1.46%) and the content of peoniflorin was 0.725 3 mg/g. Conclusions The method is accurate and reasonable, and can be used for the quality control of Wuji-Jianwei granules.
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AIM To investigate the effect of Supplemented Buyang Huanwu Decoction (Astragali Radix,Angelica tail,Paeoniae Radix rubra,etc.) on blood glucose in diabetic rats and its antioxidant activity.METHODS The diabetic rat model induced by streptozotocin (STZ) was established,with metformin as positive control group.After intragastric administration with Supplemented Buyang Huanwu Decoction,the fasting blood glucose,superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in serum and tissues (heart,kidney and pancreas) in rats were detected.HPLC was used to determine the contents of antioxidant constituents (calycosin-7-O-β-D-glucoside and peoniflorin) in plasma,whose pharmacokinetic parameters were then calculated.RESULTS Compared with the model group,the hypoglycemic activity in the Supplemented Buyang Huanwu Decoction group was obvious (P ＜ 0.05),the SOD activity in serum and various issues (except for pancreas) was significantly enhanced,together with significantly reduced MDA level (P ＜ 0.05).The pharmacokinetic behavior of two constituents (calycosin-7-O-3-D-glucoside and peoniflorin) accorded with two-compartment open model,whose blood concentrations reached the highest within 50-70 min,and showed no obvious changes within 180-720 min.CONCLUSION Supplemented Buyang Huanwu Decoction can reduce the blood glucose in diabetic rats and improve the antioxidant activities in heart and kidney.The fast absorption and slow metabolism of calycosin-7-O-3-D-glucoside and peoniflorin in decoction are beneficial to related treatment.
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This paper was aimed to explore the effects of glycosides, the effective component of Buyang Huanwu decoction, and its main active components such as astragaloside Ⅳ, amygdalin, peoniflorin and their combinations on vascular smooth muscle cells (VSMC) proliferation, clarify the major active materials of anti-VSMC proliferation and investigate the mechanisms via the signal transduction pathway. Plasma containing drug was prepared via oral administration in rats. VSMCs of rats aorta were cultured, and then VSMC proliferation was stimulated by using platelet derived growth factor (PDGF).The plasma containing drug was added to detect the activity of cell proliferation, cell cycle and related protein expressions of signaling pathway such as extracellular signal-regulated kinase (ERK), phos-phatidylinositol-3-kinase/protein kinase B (PI3K/Akt) and Janus kinase/signal transducer and activator of transcription (JAK/STAT). After being stimulated by PDGF, the proliferation activity of VSMC was strengthened (P<0.01), G₀/G₁ phase cells were decreased (P<0.01), S/M phase cells were increased (P<0.01), and PcNA, cyclin D1 protein expressions related to cell cycle were up-regulated (P<0.01). Glycosides, astragaloside Ⅳ, amygdalin, peoniflorin and their combinations could inhibit the cell proliferation (P<0.05 or P<0.01) in a dose-effect relationship and time-effect relationship. They could increase G₀/G₁ phase cells (P<0.01), decrease S/M phase cells (P<0.01), and down-regulate the protein expressions of PCNA, cyclin D1 (P<0.01); and the effects of the combinations were greater than those of single active component (P<0.05). After VSMC proliferation was induced by PDGF, p-ERK1/2 expression was increased (P<0.01), PI3K expression was down-regulated while p-PI3K expression was up-regulated (all P<0.01), and STAT3expression was reduced while p-STAT3 expression was increased (all P<0.01). Glycosides, astragaloside Ⅳ, amygdalin, peoniflorin and the combinations of these active components could reduce p-ERK1/2 expression (P<0.05), increase PI3K expression (P<0.01), decreasep-PI3K expression (P<0.05 or P<0.01), increase STAT3 expression (P<0.01), and decrease p-STAT3 expression (P<0.05 or P<0.01). These results suggested that PDGF could induce the cell cycle conversion of VSMC, leading to VSMC proliferation. The mechanism was related to the activation of ERK, PI3K/Akt and JAK/STAT signaling pathways. Glycosides and its main active components such as astragaloside Ⅳ, amygdalin, peoniflorin and their combinations can inhibit the cell cycle conversion of VSMC, with the effect against VSMC proliferation, and the mechanisms may be associated with the inhibition of PI3K/Akt, mitogen-activated protein kinase (MAPK) and JAK/STAT signaling pathways. astragaloside Ⅳ, amygdalin and peoniflorin were the major active materials of anti-VSMC proliferation, and their combination showed enhanced effect.
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Objective:To optimize the extraction parameters for the water extract of Tangganjian concentrated pills .Methods:U-sing the content of paeoniflorin and extraction yield as the evaluation indices .An HPLC was used to determine the content of peoniflorin in the extract, and the chromatographic conditions were as follows: a WondaSil C18 chromatographic column (250 mm ×4.6 mm, 5μm), the mobile phase was acetonitrile-0.1%phosphoric acid solution (16∶84) with a flow rate of 1 ml· min-1, the column tem-perature was 30℃and the detection wavelength was 230 nm.The amount of water , extraction time and extraction times were regarded as the influencing factors ,an orthogonal design was adopted to develop the analysis of variance for extraction parameters for water ex -tract.Results:The optimal extraction process was as follows:adding 12-fold amount of water and extracting 3 times with 1 h for each time.Conclusion:The optimum extraction process is reasonable , stable and feasible, which provides experimental basis for the extrac-tion process of Tangganjian concentrated pills .
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Objective To establish a quality standard for Jianpibushen granules .Methods The five main ingredients of the formulation :Astragalus membranaceus ,Salvia miltiorrhiza Bunge ,Codonopsis pilosula ,Citrus reticulata Blanco and Paeonia lactif lora Pall ,were identified by thin layer chromatography (TLC) respectively .The content of peoniflorin in Paeo-nia lactif lora was determined by high performance liquid chromatography (HPLC) .The separation was performed on Agilent Eclipse Plus C18 column(4 .6 mm × 250 mm ,5 μm) .The mobile phase was acetonitrile and 0 .1% potassium dihydrogen phos-phate solution for gradient elution .The flow rate was 1 ml/min ,the column temperature was 25 ℃ ,the UV detection was per-formed at 230 nm .Results The spots in TLC were clear without interference in negative control .A good linear relationship was shown within the range of 8 .676-277 .632 μg /ml for peoniflorin (r=0 .999 9) .Conclusion This method is simple ,accu-rate and reproducible ,and can be used as an effective quality control of Jianpibushen granules .
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OBJECTIVE:To establish a method for determining the plasma concentration of paeoniflorin and phillyrin and phar-macokinetic study before and after intragastric administration of Qianliean granules. METHODS:LC-MS/MS method was adopted. The column was Waters C18 with mobile phase consisted of acetonitrile(A)-2 mmol/L ammonium acetate(containing 0.05% formic acid)(B)(0-9 min:15%A→50%A;9-11 min:50%A→90%A;11-17 min:90%A;17-19 min:90%A→15%A;19-20 min:15%A),at the flow rate of 0.6 ml/min;column temperature was 35 ℃ and the volume was 20 μl;quantitative ions were paeoniflorin m/z 525.2 → m/z 449.0,phillyrin m/z 552.3 → m/z 355.3. 7 SD male rats were docked to collect blood 0.5 ml from angular vein 0.25,0.5,0.75,1,1.5,2,3,4,6,8,10,12,24 h after administration Qianliean granule solution 1 g(medicinal materials)/kg to determine the blood concentration of drugs. DAS 2.1.1 software was employed to calculate pharmacokinetic parameters. RE-SULTS:The linear range of paeoniflorin and phillyri were 5.0-2500.0 μg/L(r=0.9979)and 2.0-2000.0 μg/L(r=0.9982),re-spectively;RSD of precision test was less than 5.5%(n=5);the method recovery were 96.0%-104.0% and 92.0%-107.0%,the extration recovery were 71.4%-83.5% and 81.5%-92.3% and RSD of stability test was less than 5.0%(n=3). The pharmacokinet-ic parameters of paeoniflorin and phillyrin were as follows as t1/2 of (2.206 ± 0.631) and (1.355 ± 0.317) h;cmax of (1504.069 ± 620.885) and (79.043 ± 15.568)μg/L;tmax of (1.000 ± 0.250) and (1.214 ± 0.267) h;AUC0-24 h of (4897.645 ± 2207.577) and (263.475±54.795)μg·h/L;CL of(5.025±2.773)and(76.253±13.986)L/(h·kg). CONCLUSIONS:The method is highly sensi-tive,exclusive,simple,accurate and reliable,and can be applied to study the pharmacokinetic characteristics of paeoniflorin and phillyrin in rats in vivo.
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OBJECTIVE:To establish a method for the content determination of peoniflorin in Danggui shaoyao powder,and provide a reference for controlling the quality of the preparation. METHODS:HPLC was performed on the column of Symmetry C18 with mobile phase of acetonitrile-water(containing 0.1% phosphoric acid)(14∶86,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 230 nm,column temperature was 20℃,and injection volume was 20μl. RESULTS:The linear range of peoniflo-rin was 10-80 μg/ml(r=0.999 3);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 98.3%-104.9%(RSD=2.0%,n=9). CONCLUSIONS:The method is simple,accurate and specific,and can be used for the con-tent determination of peoniflorin in Danggui shaoyao powder.
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Aim To study the influence of Si-Wu De-coction (SWD ) and its active components on cyto-chrome P450 activity and mRNA expression in rats in order to provide an experimental basis for compatibility of SWD.Methods SWD and its active components were intragastrically administrated for seven days,the doses of SWD was 10 g · kg -1 · d -1 ,the doses of fructose,ferulic acid,ligustrazine,peoniflorin were 0.334,0.002,0.011 and 0.022 g·kg -1 ·d -1 ,re-spectively.After administration for seven days,rats were executed,and liver microsomes were prepared. The effects of SWD and its active components on cyto-chrome P450 in rats were investigated by hybrid probe and liver microsomes incubation method.The level of mRNA expression in liver was detected by real-time quantitative polymerase chain reaction using specific target primers for CYP450 genes.The level of protein expression of CYP2B1 was detected by Western blot. Results Compared with the control group,fructose significantly decreased the activity of CYP1A2, CYP2B6,CYP2C9,CYP2D6;ferulic acid significantly decreased the activity of CYP2C9,CYP2B6;ligus-trazine significantly decreased the activity of CYP1A2, CYP2C9,CYP2B6;peoniflorin significantly decreased the activity of CYP2D6,CYP2B6;fructose,ferulic acid,peoniflorin inhibited the mRNA expression of CYP2B1;fructose,ferulic acid,ligustrazine and peon-iflorin also inhibit the protein expression of CYP2B1. Conclusion Fructose,ferulic acid,peoniflorin inhib-it the activity of CYP2B1,decrease the expression lev-els of mRNA and protein of CYP2B1.
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To establish the quality standard for Pingyou granules. Methods: Paeoniae radix, Carthamus tinctorius, Coicis semen and Nutgrass galingale rhizome were qualitatively identified by TLC. The content of peoniflorin was determined by HPLC. The separation was performed on a Lichrospher-C18 (250 mm × 4. 6 mm, 5 μm) column with the mobile phase of methanol-acetic acid (24∶76). The detection wavelength was 232nm. Results:Paeoniae radix, Carthamus tinctorius, Coicis semen and Nutgrass galingale rhizome could be identified by TLC without any interference from the negative control. The linear range of peoniflorin was 6. 560-104. 920 μg·ml-1(r=0. 999 9) with the average recovery of 98. 77%(RSD=2. 73%,n=9). Conclusion:The qualitative identifi-cation is specific and reproducible, and the quantitative method is simple, accurate and reliable, which can be used in the quality con-trol of Pingyou granules.
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OBJECTIVE:To establish a RP-HPLC method for the determination of Peoniflorin in Tiaojingquban tablets. METHODS:The chromatographic separation was performed on VP-ODS column (150mm?4.6mm, 5?m). The mobile phase consisted of methanol-water (25∶75) at a flow rate of 1.0mL?min-1.AUV-detector at 230nm was used. RESULTS:The linear range of Peoniflorin was 24.8～198.4?g?mL-1(r=0.999 9).The mean recovery was 99.36% with RSD being 0.86% (n=6).CONCLUSION:The method is simple, rapid, sensitive and reproducible, and it can be used for the quality control of Tiaojingquban tablets.
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Objective To establish a HPLC method for determination of Peoniflorin in Zhongzi Sanda Pill.Method Kromasil-C18 column(5 ?m,4.6 mm?250 mm) was used.Mobile phase was methanol-water(30:70).The detective wavelength was at 230 nm.The flow rate was 1.0 mL/min.Results There was a good linear relationship between the injection amount 0.11～2.14 ?g and a good peak form for Peoniflorin in Zhongzi Sanda Pill.The average recovery rate was 98.2%(RSD=1.72%,n =6).Conclusion This method is convenient,effective and accurate for the quality control of Peoniflorin in Zhongzi Sanda Pill.
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Objective To optimize the extract technology of Quyou Tincture with orthogonal design.Methods With the determination of peoniflorin and paeonol and the extraction rates as indexes,the extract conditions of Quyou Tincture was optimized by orthogonal design.Results The optimal preparation process of alcohol was as follows:adding 6 times alcohol(75%),immerse 24 hours and percolate with 1 mL/min.Conclusion The optimum extracting condition was simple,with a high extraction rate and low cost.
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OBJECTIVE:To establish HPLC method for the determination of the content of paeoniflorin in Shenshao?gankang oral liquid.METHODS:Paeoniflorin was separated on Kromasil C 18 ;methanol-0.05mol/L KH 2 PO 4 solution-avantin(60∶173∶4,which was adjusted to pH=4.0with acetic acid)was taken as the mobile phase with a flow rate at1.0ml/min and detection wavelength at230nm.RESULTS:Good linear relationship was achieved when the detection concentration range of peoniflorin was0.027～0.230mg/ml(r=0.9999);The average recovery of peoniflorin was100.97%(RSD=1.28%,n=5). CONCLUSION:The method was simple,rapid,precise and reproducible,which can be used as the quality control of Shenshaogankang oral liquid.
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OBJECTIVE: To develop an HPLC method for determination of peoniflorin in Bazhen(eight ingredients) teabag.METHODS:The samples were separated on Diamonsil C18 column(250 mm?4.6 mm,5 ?m) with acetonitrile-water(17∶83) as mobile phase under a detection wavelength of 230 nm.RESULTS: The linear range for peoniflorin was 0.105 3～ 2.106 0 ?g(r=0.999 9) and its average recovery rate was 98.58%(RSD=1.17%,n=9).CONCLUSION: The method is simple,rapid,accurate and applicable for the quality control of Bazhen(eight ingredients) teabag.
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OBJECTIVE: To establish an HPLC method for simultaneous determination of peoniflorin,calycosin and ferulic acid in Kangmin granules.METHODS: The separation was performed on a Kromasil C18 column (250 mm?4.6 mm,5 ?m),and the mobile phase consisted of methanol-acetonitrile-0.4% phosphonic acid (20 ∶ 10 ∶ 70) with flow rate of 1.0 mL?min-1.The detection was performed at wavelength of 254 nm.Column temperature was room temperature and the injection volume was 10 ?L.RESULTS: The linear ranges of peoniflorin,calycosin and ferulic acid were 180～3 600 ?g?mL-1(r=0.999 2),4.6～92.0 ?g?mL-1(r=0.999 1) and 8.0～160.0 ?g?mL-1(r=0.999 4),respectively.The average recoveries were 98.3%(RSD=1.8%,n=9),99.0% (RSD=2.0%,n=9) and 100.5%(RSD=1.1%,n=9),respectively.CONCLUSION: The method is simple,accurate,sensitive and reproducible for the quality control of Kangmin granules.
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Objective To establish a method for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.Methods Reversed-phase high performance liquid chromatography(HPLC) was used.The chromatographic condition was as follows: Diamonsil C18 column(4.6mm ?250mm,5?m),column temperature being room temperature,mobile phase consisting of acetonitrile(A)-10g/L acetic acid(B) with adjusting pH value being 3.5,flowing rate at 1.0mL/min,detective wavelength at 250nm,and the injection volume being 20?L.The gradient elution procedure was as follows: elution with 18% A for 0～5 min,elution with 19%～29% A for 6～10 min,elution with 30%～45% A for 11～25min,and A+B=100%.Results Peoniflorin at the concentration of 1.916～122.6?g/L and glycyrrhizic acid at the concentration of 2.625～168?g/L showed a good linearity.The mean recovery was 101.12% for peoniflorin and 99.68% for glycyrrhizic acid.Conclusion This method is simple and reproducible,and can be used for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.
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AIM: To determine the contents of protocatechuic acid,peoniflorin,coumarin,cinnamic acid,cinnamaldehyde and glycyrrhizic acid in categorized formulas about Guizhi Decoction (Ramulus cinnamoni,Radix paeoniae alba,Rhizoma zingiberis recens,Radix et Rhizoma glycyrrhizae and Fructus jujubae) by HPLC.METHODS: The gradient elution mode was applied in chromatographic separation.The C18 column was used with the mobile phase of 0.05% phosphoric acid-acetonitrile,flow rate was at 1.0 mL/min,detection wavelength at 230 nm and 254 nm,and the column temperature was at 30 ℃.The contents of the above-mentioned six constituents were determined in Guizhi Decoction,Guizhi Decoction plus Ramulus cinnamoni and Guizhi Decoction plus Radix paeoniae alba,respectively.The differences among all combinations were tested by one-way analysis of variance using SPSS software.RESULTS: The concentrations of the above-mentioned six constituents in different decoctions were simultaneously determined by HPLC and the linear equations of six constituents were established.Compared with Ramulus Cinnamomi alone,cinnamic acid content decreased and protocatechuic acid content increased in categorized formulas about Guizhi Decoctions significantly,but then coumarin content increased in Guizhi Decoction and Guizhi Decoction plus Radix paeoniae alba.Compared with Radix Paeoniae Alba alone,peoniflorin content decreased in categorized formulas about Guizhi Decoctions significantly.Compared with Radix et Rhizoma Glycyrrhiza alone,glycyrrhizic acid content increased in categorized formulas about Guizhi Decoctions significantly.CONCLUSION:The methods are accurate,reproducible and suitable for determineation of the contents of six constituents in categorized formulas about Guizhi Decoctions,Results show that the contents change with different decoctions.