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ObjectiveTo study the differences in volatile oil content of bran-processed Atractylodes lancea and its standard decoction concentrate and freeze-dried powder， as well as the differences in the types and contents of chemical components in volatile oil， and to clarify the quality value transmitting. MethodTen batches of A. lancea rhizoma were collected and prepared into raw products and bran-processed products of A. lancea， standard decoction concentrate and freeze-dried powder of bran-processed A. lancea in order to extract the volatile oil， and the transfer rate of volatile oil in each sample was calculated. Quantitative analysis of the main chemical components（β-eudesmol， atractylon， atractylodin） in each volatile oil was performed by gas chromatography（GC） on the HP-5 quartz capillary column（0.32 mm×30 m， 0.25 μm） with a flame ionization detector（FID）， a split ratio of 10∶1 and a temperature program（initial temperature at 80 ℃， hold for 1 min， rise to 150 ℃ at 10 ℃·min-1， hold for 10 min， rise to 155 ℃ at 0.5 ℃·min-1， hold for 5 min， rise to 240 ℃ at 8.5 ℃·min-1， hold for 8 min）. Cluster analysis and principal component analysis（PCA） were used to explore the overall differences in types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ResultThe transfer rates of volatile oil in the bran-processed products， standard decoction concentrate and freeze-dried powder were 70.51%， 1.57% and 40.90%， respectively. The average transfer rates of β-eudesmol， atractylon and atractylodin in the volatile oil of bran-processed A. lancea were 58.45%， 48.49% and 55.64%， respectively. In the standard decoction concentrate， only β-eudesmol and atractylodin were detected， and their average transfer rates were 0.22% and 0.10%， respectively. And only β-eudesmol was detected in the freeze-dried powder with the average transfer rate of 8.37%. The results of cluster analysis and PCA showed that there are obvious differences in the types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ConclusionThe quality value transmitting between bran-processed A. lancea and its standard decoction concentrate and freeze-dried powder is stable， and if the freeze-dried powder is selected as the reference material of dispensing granules， appropriate amount of volatile oil should be added back to make it consistent with the quality of the standard decoction concentrate.
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The aim of this paper was to explore the effect of Xueshuantong Injection(freeze-dried powder,XST) on κ-carrageenan-induced thrombosis and blood flow from the aspects of interactions among blood flow,vascular endothelium and platelets. Fifty male Sprague-Dawley rats(190-200 g) were randomized into five groups: control group, model group, heparin sodium(1 000 U·kg~(-1)) group, low-dose and high-dose(50, 150 mg·kg~(-1)) XST groups. Rats were intraperitoneally injected with corresponding drugs and normal saline(normal control and model groups) for 10 days. One hour after drugs were administered intraperitoneally on the 7 th day, each rat was injected with κ-carrageenan(Type Ⅰ, 1 mg·kg~(-1)) which was dissolved in physiological saline by intravenous administration in the tail to establish tail thrombus model. The lengths of black tails of the rats were measured at 2, 6, 24 and 48 h after modeling. Vevo®2100 small animal ultrasound imaging system was used to detect the internal diameter of rat common carotid artery, blood flow velocity and heart rate, and then the blood flow and shear rate were calculated. Meanwhile, the microcirculatory blood flow perfusion in the thigh surface and tail of rats were detected by laser speckle blood flow imaging system. Platelet aggregometry was used to detect the max platelet aggregation rate in rats. Pathological changes in tail were observed through hematoxylin-eosin staining, and Western blot was used to detect the protein content of platelet piezo1. According to the results, XST could inhibit the rat tail arterial thrombosis and significantly reduce the length of black tail(P<0.05). The blood flow of common carotid artery in XST low dose group was significantly higher than that in the model group(P<0.05). XST high dose group could significantly increase the microcirculatory blood flow perfusion of the tail in rats as compared with the model group(P<0.05). XST high dose group could significantly inhibit platelet aggregation rate(P<0.05) and XST low dose group could significantly inhibit platelet piezo1 protein expression(P<0.01). In summary, XST could play an effect in fighting against thrombosis induced by κ-carrageenan in rats, which may be related to significantly inhibiting platelet aggregation, improving body's blood flow state, maintaining normal hemodynamic environment and affecting mechanical ion channel protein piezo1.
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Animals , Male , Rats , Drugs, Chinese Herbal , Microcirculation , Rats, Sprague-Dawley , ThrombosisABSTRACT
Objective:To study on the pharmacokinetics and tissue distribution of baicalin magnesium salt in rats after tail vein injection,and compare pharmacokinetic differences between baicalin magnesium salt and baicalin. Method:After tail vein injection of baicalin magnesium salt and baicalin,orbital blood was collected at different time points.The drug concentration was measured by HPLC,the drug concentration-time curve was plotted,the pharmacokinetic parameters were calculated with DAS 3.0 software,SPSS 19.0 software was used for statistical analysis.At the same time,the drug distribution in heart,liver,spleen,lung and kidney was measured at different time points after tail vein injection of baicalin magnesium salt. Result:When the dose of baicalin magnesium salt was 25-100 mg·kg-1,area under the curve(AUC0-t and AUC0-∞) showed a good linear relationship with the dose(r>0.95),but most of the other pharmacokinetic parameters had no significant difference between different dose groups.The mean residence time(MRT0-t) of medium dose group of baicalin magnesium salt was significantly higher than that of equal molar dose group of baicalin.After intravenous injection of baicalin magnesium salt,the drug concentration was the highest in each tissue at 0.25 h,and the concentration of target component decreased rapidly at 0.75 h.The distribution of target component in kidney was the most,followed by lung. Conclusion:After injection,the baicalin magnesium salt can be rapidly distributed and quickly eliminated in vivo,which is mainly excreted from the kidney.
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Objective To study the effects of Jiedu Quyu Ziyin Prescription (JQZP)-treated freeze dried powder and drug-containing serum on the inflammatory signal pathway of monocyte-macrophage induced by LPS (lipopolysaccharide) in mice. Methods Monocyte-macrophage cells were cultured in vitro and randomly divided into blank group, LPS stimulation group, drug-containing serum group, freeze dried powder group, LPS + drug-containing serum group, and LPS + freeze dried powder group. After 24 h intervention, the optimal concentrations of drug-containing serum and freeze dried powder were screened by CCK8 method and the cell viability was measured respectively. The content of tumor necrosis factor (TNF-α) in cell serum was measured by ELISA. Real-time PCR was employed to test the expression of TNF-α mRNA and nuclear transcription factor kappa-light-chain-enhancer of activated B cells (NF-κB). Western-blot was used to detect the expression of NF-κB protein. The LC-MS was used to detect the active ingredients in the drug-containing serum. Results Compared with the blank group, the expression of TNF-α level, NF-κB, TNF-α mRNA and NF-κB protein in LPS stimulation group were significantly increased (P < 0.05). Compared with the LPS stimulation group, the TNF-α level, NF-κB, TNF-α mRNA and the expression of NF-κB protein in the LPS plus serum group were significantly lower than those in the LPS plus freeze-dried powder group (P < 0.05). Paeoniflorin and ferulic acid were detected in the drug-containing serum. Conclusion JQZP freeze-dried powder and drug-containing serum all have the effect of inhibiting the inflammatory signaling pathway.
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OBJECTIVE:To establish the quality standard of Hirudo nipponica freeze-dried powder(called"freeze-dried powder"for short),and to provide reference for controlling its quality. METHODS:A total of 3 batches of freeze-dried powder were collected,identified and tested according to the requirements of H. nipponica stated in 2015 edition of Chinese Pharmacopoeia(part Ⅰ)(shorted for pharmacopoeia);the antithrombin activity was also analyzed. The maximum tolerated dose (MTD)was used to investigate the toxicity. The stability was determined by designing temperature,humidity and strong light exposure tests. RESULTS:In the TLC of test sample,the same red spots were found in the corresponding location of the control drug chromatogram,and the same orange-red fluorescence spots were shown under the UV light(365 nm). Average content of moisture in 3 batches of samples was 2.61%,and the levels of total ash,acid-insoluble ash,pH aflatoxin and antithrombin activity were 2.83%,0.38%,6.92,0.28 μg/kg and 257.0 U/g,respectively. The content of Pb,Cd,As and Cu were in line with the requirements of pharmacopoeia except that the content of Hg was slightly higher than lower limit of H. nipponica in pharmacopoeia. Results of MTD showed that no death and ADR was found in mice after giving 26.4 g/kg freeze-dried powder by the amount of crude drug,which was 58 times as large as the maximum dosage that the pharmacopoeia described. Under the condition of 20, 40 ℃ and strong light exposure [(4 500±500)Lx],the anticoagulase activity of freeze-dried powder decreased significantly over time,while the anticoagulase activity of freeze-dried powder stored at 40 ℃ for 6 months was in line with the requirements of pharmacopoeia. Under the condition of high humidity(relative humidity were 90%,75%),freeze-dried powder showed a strong hygroscopicity. CONCLUSIONS:Established quality evaluation standard for freeze-dried powder according to pharmacopoeia standard could be used to control its quality.
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AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.
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OBJECTIVE:To prepare the Albendazole nanoliposomes freeze-dried power and study its properties. METHODS:Freeze-drying method was conducted to prepare Albendazole nanoliposomes freeze-dried power,using the particle size,encapsula-tion efficiency,appearance,redispersibility as indexes,single factor test was combined with orthogonal test to screen freeze-drying preparation technology. The morphological changes,particle size,Zeta potential,moisture content,12 months stability at 4 ℃ be-fore and after freeze-drying were detected. RESULTS:Plus a total content of freeze-dried protective agent was 10%,the ratio of glucose-trehalose-mannitol was 1.0:1.0:3.0,using quick-freeze,pre-freezing 18 h in -35 ℃ refrigerator,dry-freezing 48 h to ob-tain freeze-dried powder. Compared with before freeze-drying,the freeze-dried liposomal morphology had no obvious changes, showing clear phospholipid bilayer membrane structure;the particle sizes before and after freeze-drying were (208.63 ± 1.04) nm and (223.04 ± 2.02) nm,Zeta potentials were (-15.6 ± 0.04) mV and (-19.4 ± 0.06) mV,encapsulation efficiencies were (94.62±0.49)%and(91.10±0.46)%(n=3),respectively. Compared with liposomes,liposomes freeze-dried power had good sta-bility in 12 months at 4 ℃. CONCLUSIONS:Albendazole nanoliposomes freeze-dried power is prepared successfully,its stability is superior to albendazole nanoliposomes,and the freeze-drying technology is feasible.
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Objective: To study the correlation between the empty bottle volume, negative pressure and gas production of the freeze-dried powder in the out-patient pharmacy intravenous admixture center of a children' s hospital in order to provide reference for the drug production. Methods:20 ml Syringes were used to measure the volume of empty bottles, negative pressure and produced gas. The relationship between the theoretical drug dissolution volume and the actual dissolution volume was compared, and the precautions for the drug production were put forward. Results:Among the tested 30 drugs, 6 ones were with the actual dissolution volume half of the theoretical dissolution volume, 8 ones were with negative pressure in the bottles, and 3 ones were with produced gas after dissol-ving. It was appropriate that the empty bottle volume be 4 ml larger than the theoretical dissolution volume, and it was appropriate that the negative pressure volume of drugs was slightly larger than the theoretical dissolution volume. Negative pressure should be still kept in the bottles after the gas production. Conclusion:The design of part of freeze-dried powder injection needle shows defects resulting in drug mixing difficulties to a certain extent.
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OBJECTIVE:To establish a method for the simultaneous determination of the contents of Shuanghuanglian freeze-dried powder in placental perfusate. METHODS:HPLC was performed on the column of Agilent Zorbax-C18 with mobile phase of acetonitrile-1%formic acid aqueous solution(gradient elution)at flow rate of 1.0 ml/min,the internal standard was puera-rin,detection wavelength was 280 nm,column temperature was 25 ℃ and sample volume was 10 μl. RESULTS:There was a lin-ear range between linear ranges and peak area of 8 ingredients(r≥0.999 0);RSDs of within-day and inter-day precision tests were no more than 1.9%,repeatability tests was no more than 7.3%;average recoveries were in the range of 92.73%-112.37%(RSD=3.2%-8.2%,n=6);and average matrix effects were 90.33%-105.78%(RSD=3.2%-8.0%,n=6). CONCLUSIONS:The method is rapid,sensitive and specific,and can be used to the simultaneous determination of the contents of 8 ingredients of Shuan-ghuanglian freeze-dried powder in placental perfusate.
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Aims: With pickled perilla leaves as raw materials, this paper proposed the optimal ethanol extraction conditions and made a profound analysis for extract in the compositions of major active ingredients, nutrients, mineral elements and amino acid to characterize the nutritional and biological properties of pickled perilla leaves, which could aid its finely processing and future application in the development of functional food. Methods: The optimum ethanol extraction process for preparing freeze-dried powder from pickled perilla was studied by means of orthogonal experiments, with the concentration of ethanol, extracting temperature and extracting time as factors. Meanwhile, the contents of the activity components, such as polysaccharide, flavones and rosmarinic acid, as well as the mineral elements and nutritional contents in the freezedried powder were determined according to the methods reported by relevant literatures without or with a few modifications. Results: The optimal extracting conditions as follows: 50ºC of temperature, 60 min of extraction time and 80% of ethanol concentration. Under the optimal extracting conditions, the extraction rate of the freeze-dried powder was 1.71%. Moreover, perilla leaf extract contained rich biological and nutritional ingredients, including 33.39% of flavonoids, 9.24% of polysaccharides, 22.79% of rosmarinic acid, 5.47% of protein, 7.61% of fat, 2354 mg/kg of Ca, 111.4 mg/kg of Fe, 5.045 mg/kg of Zn, 1817 mg/kg of K , 12.66 mg/kg of Mn and nine of essential amino acids. In addition, perilla leaf extract exhibited obvious scavenging effects on the DPPH•, •OH and O2 •. Conclusion: In summary, pickled perilla leaf ethanol extract was rich in biological ingredients as well as a variety of nutrients, and showed antioxidant activities in vitro, thus it is valuable and promising in the development of functional foods in the future.
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Objective To investigate Anti-aging function of Placenta freeze-dried powder on mice. Method 2 month old female-KM mice were divided into five groups:normal control group, aging model group, positive control group, placenta freeze-dried powder low dose group and high dose group. Except normal control group,the rest of groups were treated with method of subcutaneous continuous injection of D-galactose for 42 days in order to establish mice aging model. Meanwhile, the corresponding drugs, via intragastric administration, were ingested by different treated groups and observe the change of immunity and physiological regulation related in mice. Results Compared with aging model group, thymus index and spleen index in placenta freeze-dried powder low dose group and high dose group increased obviously (P<0.05), content of MDA in brain decreased significantly (P<0.05), proportion of CD 4 and CD 8 lymphocyte in total lymphocyte, female hormone(P and E 2), IgG and haemopoietic factor in peripheral blood increased remarkably. Conclusion Placenta freeze-dried powder could slow the aging process on mice via immunity enhancement and improving physical regulation.
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OBJECTIVE: To investigate the pharmacokinetics of puerarin of Zige freeze-dried powder in the plasma of the normal ruts and the model rats with middle cerebral artery occlusion(MACO). METHODS: A high-performance liquid chromatography-tan-dem mass spectrometry (HPLC-MS/MS) method was developed and validated for quantification of puerarin in rat plasma after injection administration of Zige freeze-dried powder. The plasma samples were pretreated with methanol for protein precipitation and the supernatants were analysis on a Diamonsil C18 column (4.6 mm × 150 mm, 5 μm) with the mobile phase consisting of 0.1% formic acid in methanol and 10 mmol · L-1 ammonium acetate in water (80:20) at a flow rate of 0. 6 mL · min-1 to separate puerarin and genistein (internal standard, IS). A triple-quadrupole tandem mass spectrometer under a multiple reaction-monitoring (MRM) mode to determine puerarin(m/z 415.1 → 295.1) and genistein(m/z 269.2 → 133.1). The Zige freeze-dried powder was administered to rats in 26.7 mg · kg-1 doses via intravenous (iv) injection, the plasma sample was collected at each time point (3, 5, 10, 30 min, 1, 2, 3 and 5 h post-administration). The pharmacokinetic analyses of puerarin in rat plasma were performed via the proprietary DAS 2.1 computer software package. RESULTS: The calibration curves had good linear in 0.02-100 μg · mL-1 with r=0.9954. The intra- and inter day precisions were found to be less than 13.2%, the accuracy was ranged in -7.3%-10.5%, and the extraction recoveries of the puerarin from the plasma was 86.3%. The contents of puerarin arrived the max values with (56.5 ± 3.4) and (62.3 ± 14.0) μg · mL-1, the t1/2 were (0.311 ± 0.133) and (0.755 ± 0.128) h, the values of AUG0-t, were (17.1 ± 1.5) and (20.0 ± 6.8) μg · h · mL-1, the values of MRT0-t, were (0.269 ± 0.044) and (0.360 ± 0.045) h in the plasma of the normal and the model respectively. Compared with the normal, the t1/2, MRT0-t, of the model were significantly longer (P < 0.05). CONCLUSION: The developed and validated method is specifically, rapid and sensitive, and has been successfully applied to research the pharmacokinetics of puerarin in rats plasma after iv zige freeze-dried powder. The pharmacokinetic; parameters showed that the puerarin in the Zige freeze-dried powder has better biological availability, slower elimination and longer resistance time in the model rats, which indicate the reduce of the rate of metabolism in the model and is beneficial to the clinical pharmacokinetic studies after iv administration of Zige freeze-dried powder.
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Objective To study the general pharmacological effects of Aloe's whole-leaf freeze-dried powder (AWFD), and observe its influence on cardiovascular system, nervous system and respiratory system of laboratory animals, so as to offer an experimental basis for clinical application. Methods Forty-eight mice were randomized into blank control group, high dosage group, medium dosage group and low dosage group of AWFD (12 mice for each group). AWFD high, medium and low dosage groups were treated by intragastric at the dose of 12.20, 3.90, 0.65 g/(kg?d), blank control group was treated by equal volume of sterilized distilled water. After three days, general behavior, spontaneous activity, coordinated movement, sleep situation induced by sodium pentobarbital in subthreshold dose and suprathreshold dose were observed. Twenty-four beagle were randomized into blank control group, high dosage group, medium dosage group and low dosage group of AWFD (6 beagles for each group). AWFD high, medium and low dosage groups were treated by duodenum at the dose of 6.10, 3.41, 0.71 g/(kg?d), blank control group was treated by equal volume of sterilized distilled water. The influence on blood pressure, heart rate, electrocardiogram, breathing flow and frequency in anesthetic dogs were observed. Results Three dosages of AWFD had no obvious influence on spontaneous activity and coordinated movement in mice, and had no evidently influence on sleep number and duration, but the high dosage group of AWFD had influence on sleep latency (P<0.01). AWFD had no impact on blood pressure, heart rate, electrocardiogram, breathing flow and frequency in anesthetic dogs. Conclusion AWFD has no evident effects on cardiovascular system and respiratory system in laboratory animal, however, the impact on the central nervous system remains to be further verified.
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Objective To observe the effect of Yinhuang freeze-dried powder injection on nervous system of rats and mice,and to probe its safety.Methods After injected Yinghuang freeze-dried powder injection nerve system and general behavior of rat or mouse were observed.Results Yinghuang freeze-dried powder didn’t affect rat’s or mouse’s general behavior,but had certain central nevous system suppression function in rats and mice.Conclusion Yinghuang freeze-dried powder have certain central nevous system suppression function in rats and mice.
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Objective To observe the rat chronic toxicity induced by Yinghuang freeze-dried powder injection. Methods The rats were given Yinghuang freeze-dried powder injection for 35 days with the doses of 166.5, 82.25, 41.12 mg/kg and for 15 days of stopping administration. The index sign of hematology and biochemistry were determined, and the pathological examination were carried out. Results The rat were given Yinghuang freeze-dried powder injection for 35 days and for 15 days of stopping administration. Mostly index sign of hematology and biochemistry had no significant difference by comparing with the control. There exceptional index sign of exceptional dosage group have significant difference by comparing with the control. But these exceptional index sign fluctuated in normal range in the laboratory. The pathological examination did not discover significant pathological changes related to drug toxicity. Conclusion Yinghuang freeze-dried powder injection has no significant toxicity for long-term administration on rats. It infers that the drawing up doses in clinic will be safe.
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Object To prove the feasibility of bacterial endotoxin test (BET) in Shenmai Freeze-dried Powder Injection by interference test. Methods Interference primary screening test and interference test were conducted in BET of Shenmai Freeze-dried Powder Injection to detect whether the interference existed or not and to explore the method of removing the interference. Results There was inhibition on BET with 0.5 EU/mL tachypleus amebocyte lysate (TAL). Interference could be excluded by using 0.25 EU/mL TAL and diluting the samples. Conclusion It is feasible to use 0.25 EU/mL TAL or TAL with more sensitivity in BET of Shenmai Freeze-dried Powder Injection.
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AIM:The different factors on stability of salvianolic acid B in Danshen freeze-dried powder injection was investigated to provide the experimental data for clinical application and storage of the preparation. METHODS: The stability of salvianolic acid B for lamplight,temperature,sodium chloride injection and glucose injection in Danshen freeze-dried powder injection were studied and the content of salvianolic acid B was determined by HPLC. RESULTS: The content of salvianolic acid B in Danshen freeze-dried powder injection didn't almost change under the condition of lamplight(3000lx) in 60 d.Its expiry date estimated by Q_(10) method was(2.39) a.The preparation was matched respectively with(0.9%) sodium chloride injection and 5% glucose injection,the content of salvianolic acid B was invariable and insoluble particles were qualified under the condition of 25 and 37 ℃ in 24 h. CONCLUSION: Danshen freeze-dried powder injection is stable for lamplight and clinical injection and the expiry data of the preparation is(2.39) a.