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Objective:Real-time cell-based assay (RTCA) technology was used to detect the influence of compound Danshen tablets with different concentrations on cells to screen outthe sensitive, stable and concentration-dependent cell lines to provide reference for the quality evaluation experiment of cell biological effects of compound salvia miltiorrhiza tablets. Methods:The effects of compound Danshen tablets on human umbilical vein endothelial cells (Huvec), rat cardiac myocytes (H9C2) and rat thoracic aortic smooth muscle cells (RA-VSMC) were investigated by RTCA. The cell index (CI) was measured to form time-dose dependent cell response curves (TCRPs) to screen the the optimal cell lines by description analysis and statistical analysis. Results:All of the three cells show obvious response to the effects of compound Danshen tablets. The sensitivity of RA-VSMC was relatively poor, the range of effective concentrationof three cells was different, and the concentration gradient dependence was also different. H9c2 cells had good sensitivity to compound Danshen tablets, which was gradient dependent, and the effective concentration range wass 0.3-1.8 mg/ml, and it can be used as a cell for quality evaluation of compound Danshen tablets. Conclusions:RTCA technology can monitor and record the growth status of cells in real time. It can accurately reflect the effects of compound Danshen tablets on different cell lines. The data are objective and reliable. It provides a new experimental technology and method for the quality evaluation of compound Danshen tablets.
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OBJECTIVE:To establish a method for simultaneous determination of hyperoside,quercitrin,luteoloside, kaempferol,quercetin,rutin,luteolin and isorhamnetin in total flavanones of Sedum sarmentosum Bunge. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ZOBAX SB C18column with mobile phase consisted of methanol-5 mmol/L ammonium formate aqueous solution(45:55,V/V)at the flow rate of 0.4 mL/min. The column temperature was 30 ℃, and sample size was 2 μL. The electrospray ionization source(ESI)was used;ion source temperature was 400 ℃;desolvation temperature was 300 ℃;desolvation gas flow was 600 L/h;capillary voltage was 3 000 V;nebuliser pressure was 45 psi;the work mode was multiple reaction monitoring mode;detection mode was negative ion mode. The established method was used to determine the contents of 8 components in 3 batches of total flavanones of Sedum sarmentosum Bunge. RESULTS:The linear ranges of hyperoside,quercitrin,luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin were 10.0-640.0,0.5-32.0, 4.5-288.0,8.0-512.0,50.0-3 200.0,2.0-128.0,12.5-800.0 and 25.2-1 612.8 ng/mL(r≥0.991 4),respectively. The limits of detection were 5.0,0.25,2.25,4.0,25.0,1.0,6.25 and 12.6 ng/mL,respectively.The limits of quantitation were 10.0,0.5,4.5, 8.0,50.0,2.0,12.5 and 25.2 ng/mL,separately. RSDs of precision,stability(24 h)and reproducibility tests were no more than 4.3%(n=6). The recoveries were 95.9%-100.6%,and RSDs were 1.5%-3.8%(n=6). The contents of hyperoside,quercitrin, luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin in 3 batches of total flavanones of Sedum sarmentosum Bunge were 507.88-560.37,42.95-50.36,63.52-71.80,1 695.10-1 753.27,10 569.28-10 612.99,25.76-30.13,2 795.22-2 877.43 and 4 869.55-4 971.30 μg/g,respectively. CONCLUSIONS:The established method can be used for simultaneous determination of 8 components in total flavanones of Sedum sarmentosum Bunge.
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Objective To develop an UPLC-MS/MS method for simultaneously determination of five components (adenosine,cytidine,guanosine,mannitol and adenine) in Qiangshenpaidu capsules. Methods The UPLC separation was performed on an Agilent ZOBAX SB-C18(2.1 mm×150 mm,5 μm) column.Isocratic elution was carried out with mobile phase consisting of methanol-0.1%formic acid (5∶95) at a flow rate of 0.2 mL.min-1.The mass spectrometer was operated in the positive ionization electrospray ( ESI) mode using multiple monitoring ( MRM) for analysis of five components. Results Adenosine,cytidine,guanosine,mannitol and adenine were all analyzed with good precision and accuracy. The linear ranges were 35-1 120 ng.mL-1( r=0.998 1) ,10-320 ng.mL-1( r=0.996 4) , 30-980 ng.mL-1( r=0.999 3) , 40-1 280 ng.mL-1( r=0.993 4), 25-800 ng.mL-1(r=0.996 5),respectively.The recoveries of six analytes ranged from 97.4% to 103.6% and the relative standard deviations were all below 4.7%. Conclusion A sensitive,accurate and suitable UPLC-MS/MS method has been developed,and the method could be applied for the determination of adenosine,cytidine,guanosine,mannitol,and adenine in Qiangshenpaidu Capsules.
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Objective: To study the acute and long-term toxicity of brucea javanica oil subnanoemulsion injections ( BJOSI ) . Methods:The mice were given BJOSI by intravenous injection. The acute toxicity was observed and LD50 in mice was calculated by Bliss method. To observe the long-term toxicological effects, Beagle dogs were injected intravenously BJOSI once a day for 8-week du-ration with the dose of 20, 10 and 6 ml·kg-1 , respectively followed by 3-week recovery period. Results:LD50 of BJOSI in mice was 7. 388 g·kg-1 with 95% confidence limits of 6. 306-8. 656 g·kg-1 . The long term toxicity test showed that all the detected indices were within normal range in all groups, including general state, weight changes,hematological indices,biochemical indices,EEG,organ coefficients, morphological and histological changes, while there was an upward tendency of ALT and Cr in every BJOSI group without dose-effect correlation. The dogs could completely recover in three weeks after the administration. Conclusion:The results suggest that BJOSI has low toxicity,while the pathological changes are reversible, and attention should be paid to the function of liver and kidney.
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Objective To choose the optimum extraction process for capsule of Buyang Huanwu.Methods With the orthogonal method,the main factors effecting the water-extraction process,such as the quantity of water,the time,pH and degree for decoction,were reviewed according the content of total glycoside and Astragalus Glycoside A.Results The optimum condition of water-extraction was extracting with 8 times amount of water for the first time and 4 times amount for the second and third time,1 hour for each time,pH was 12.Conclution The experimental results provide the basis for the water extraction process for capsule of Buyang Huanwu.
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Objective To research the content changes of Aristolochic Acid-A from Caulis Aristolochiae Manshuriensis and its processed products.Method Chromatographic assay was performed on Lichrospher-C18 column (4.6 mm?200 mm,5 ?m) with methanol-0.1% glacial acetic acid solution (70∶30) as mobile phase,the flow rate was 1.0 mL/min and the detection wavelength was set at 310 nm,the column temperature was 30 ℃.Result The content of Aristolochic Acid-A was lower in three processed products than in crude drugs,and the reduction rate of the one which was boiled by NaHCO3 was the highest.Conclusion The three processed method can reduce the content of Aristolochic Acid-A,and achieve the aim of reducing the toxicity.
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Objective To establish quality control method for Qingzao Runfei Mixture. Methods Mulberry Leaves,Glycyrrhiza,Radix Glehniae,Ophiopogon Japonicus and Loquat Leaves were identified by TLC. Glycyrrhizic Acid was determinated by HPLC. Results The negative sample of TLC had no interference. The specifity was good. Glycyrrhizic acid showed a good linear relationship in the range of 0.203 1～1.692 1 ?g,r =0.999 8. The average recovery was 98.24%,and RSD was 2.3%. Conclusion The established methods are simple,quick and with good reproducibility. This study provides methods for the quality control of Qingzao Runfei Mixture.
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Objective To establish the quality standard for Kangyuan Granula. Methods Radix Astragali, Radix Paeoniae Alba, Fructus Lycii and Radix et Rhizoma Glycyrrhizae in this prescription were identified by TLC. The content of the Paeoniflorin was determined by HPLC. The column of AichromBond-AQ-C18 (4.6 mm?150 mm, 5 ?m) was used, the mobile phase was Water-Acetonitrile (85∶15), at the flow of 1.0 mL/min, the column temperature of 30 ℃, and peaks were detected at 230 nm. Results The characteristic of identification by TLC was distinct and highly specific. The linear range of Paeoniflorin was 0.0545～0.545 ?g, r =0.999 5, the average recovery was 97.90%, RSD=1.06% (n =6). Conclusion The method was easy to operate with accurate result and good reproducibility. It can be used effectively for the quality control of this preparation.
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Objective To establish the quality standard for Kangganmao Granule.Methods Radix scutellariae,radix et rhizome glycyrrhizae and radix platycodonis in the prescription were identified by TLC.The content of the baicalin was determined by HPLC.The column of AichromBond-AQ-C18(4.6 mm? 150 mm,5 ?m)was used.The mobile phase was CH3OH-H2O-H3PO4(43:57:0.2),with the flow of 1.0 mL/min,the column temperature of 30 ℃,and peaks were detected at 280 nm.Results The characteristic of identification by TLC was distinct and highly specific.The linear range of baicalin was 0.049 6～0.297 6 ?g,r =0.999 7,the average recovery was 101.80%,RSD=1.86%(n=6).Conclusion The method was easy to operate with accurate result and good reproducibility.It can be used effectively for the quality control of the preparation.
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Mangiferin, one of the active constituents of Rhizoma Belamcandae, in samples of Be-lamcanda chinensis (L.) DC. or its substitute was determined quantitatively by RP-HPLC. The 11 sam-ples collected from different localities for analysis were: 7 rhizomes of wildly grown or cultivated B. chi-nensis, 1 of its leaf and stem, and 3 substitutes (a wildly grown and another commercially available Iristectorum Maxim. and a I. dichotoma Pall. ). Results of the analysis showed that the contents of mangiferinin Rhizoma Belamcandae were significantly higher than that of its substitutes I. tectorum and I. di-chotoma. There were also certain significant differences between samples from different localities (P<0.05), but with no statistically significant difference between the rhizome or leaf and stem, neither be-tween cultivated and wildly grown samples, (P>0.05). The method was proved to be quick, simple andreproducible, and may provide a reliable basis for the quality control and evaluation of B. chinensis.
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0.99.The quantitive restriction was 0.1 ?g/mL,the recovery rate was over 70%.The RSD of intra-and inter-day was less than 15%.The pharmacokinetic parameters of flavonoids were different from each other after ig and iv administration.The absolute bioavailability of flavonoids were 12.9%,10.8%,2.2%,10.2% and 5.9%,respectively.Conclusion The method is sensitive,specific,accurate,and is not only useful for guiding the bioavailability study on the corolla of A. manihot,but also for establishing a reasonable clinical dosage program.The four flavonols in the corolla of A.manihot can be rapidly distributed and eliminated in rats,the pharmacokinetics of two routes of administration are different.A novel integrated pharmacokinetic approach to describing the holistic pharmacokinetic properties of Chinese materia medica has been successfully developed and validated using four flavonols of A.manihot as a model herbal medicine.This study would be a new try for guiding the holistic pharmacokinetic study in consistence with the intrinsic theory and characteristics of traditional Chinese medicine.
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Objective To establish a method for the determination of Baicalin content in Shengjijiedu Granule by HPLC.Method HPLC was performed to determine naringin content on C18 column.The mobile phase consisted of a mixture of CH3OH-H2O-H3PO4(50 ∶50 ∶0.2) and detection wavelength was at 280 nm.Result The linearity of baicalin was in the range of 0.015～0.121 ?g (r=0.9999) and the average recovery rate was 100.98 %,RSD=0.84 %.Conclusion This method was simple,sensitive and accurate,and can be used for the quality control of Shengjijiedu granule.
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AIM:To establish the method for quality control of Suhong Tongluo Tincture(eugenol and methylsalicylate). METHODS: Gas phase chromatogram approach was used to determine the content of eugenol and(methylsalicylate.) The chromatogram condition consisted of 10% PEG-20M,carrier was N2(99.999%),temperature of the entrance of the column was at 230(?C),temperature of the detector was at 250(?C);column temperature was at 70(?C)(4 min),3(?C)/min→90(?C)(4 min),then 13(?C)/min→200(?C)(3 min);0.5 ?L Sample quantity. RESULTS: The linear range of methylsalicylate was 2.41-14.21 ?g,r=0.999 7,the average recovery was (99.76%),RSD=(2.64%(n=6).) The linear range of eugenol was 2.43-14.34 ?g,r=0.999 96,the average recovery was (99.33%),RSD=2.47%(n=6). CONCLUSION: This method is simple,quick,accurate and reliable for the quality control of Suhong Tongluo Tincture.