Screening and optimization of the formulation of human recombinant neutrophil inhibitory factor and hirulog hybrid for injection / 中国生物制品学杂志

@#Objective To screen and optimize the formulation and technology of human recombinant neutrophil inhibitory factor and hirulog hybrid(TNHH)for injection,and investigate its stability.Methods Based on the results of the single factor experiment,with the pH range,mannitol dosage and povidone K30 dosage as independent variables,and the content of high molecular protein as response value,the response surface design(CCF)test was used to analyze the effects of the respective variables and their interaction on the content of high molecular protein in TNHH for injection to screen out the optimal formulation. In order to facilitate the operation,the optimal formulation was adjusted to prepare three batches of samples in pilot scale,which were placed at 40 ℃,75% relative humidity(RH)for 2,4 weeks and 2 ～ 8 ℃ for 3,6 months,respectively. The samples were taken and the appearance,pH,purity of reversed phase-high performance liquid chromatography(RP-HPLC)and purity of size exclusion chromatography-high performance liquid chromatography(SECHPLC)were detected to verify the stability of this formulation and process.Results The optimal formulation was pH 4. 982 6,mannitol 7. 986 4% and povidone K30 1. 902 7%,which was finally adjusted to pH 5. 0,mannitol 8. 0% and povidone K302. 0%. The TNHH preparation for injection prepared by the optimized prescription and process were stable in quality and met the clinical medication requirements.Conclusion The optimum formulation of TNHH preparation for injection is reasonable in the process and suitable for industrial production.

Chemical Composition Analysis and Characterization of Reference Sample of Huangqi Guizhi Wuwutang Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

ObjectiveTo analyze the chemical composition of the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder）， and to provide quality markers for the formulation of quality standards of this formula. MethodUltra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS） was performed on a Waters ACQUITY UPLC™ HSS T3 column （2.1 mm×100 mm， 1.8 μm）， the mobile phase was methanol （A） -0.1% formic acid aqueous solution （B） for gradient elution （0-8 min， 1%-20%A； 8-10 min， 20%-30%A； 10-12 min， 30%-35%A； 12-14 min， 35%-40%A， 14-23 min， 40%-55%A， 23-27 min， 55%-99%A； 27-28 min， 99%A； 28-28.5 min， 99%-1%A； 28.5-30 min， 1%A）， the column temperature was 40 ℃， the injection volume was 2 μL， and the flow rate was 0.3 mL·min-1. The mass spectrometry data of the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder） were collected under positive and negative ion modes. The conditions of mass spectrometry were electrospray ionization （ESI）， scanning range of m/z 50-1 200， and impact energy of 10-30 eV. UNIFI 1.8 and Progenesis QI 2.0 software were used to analyze and characterize the chemical constituents in reference sample of Huangqi Guizhi Wuwutang （lyophilized powder） combined with reference comparison and literature review. ResultA total of 123 chemical constituents were identified， including 33 flavonoids， 26 glycosides， 18 organic acids， 11 terpenoids， 7 phenylpropanoids， 4 gingerol， 3 alkaloids， 3 amino acids， 2 amides and 16 other compounds. ConclusionThe established method can quickly and accurately characterize the chemical components in the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder）， which can provide a basis for the selection of quality evaluation indicators of this formula， and provide a reference for its preparation research.

Biological Evaluation for Quality Control of Water Extract of Qingjin Huatantang Based on Phagocytic and Secretory Functions of Macrophages / 中国实验方剂学杂志

Objective:To establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages， and to control the quality of this formula from the biological activity level. Method:The phagocytic and inflammation models of RAW264.7 macrophages were established， the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6 （IL-6） secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang， and the biological limit was searched. Result:The optimal inoculation density of RAW264.7 macrophages was 3×10⁵ pcs/mL， and the concentration of lipopolysaccharide （LPS） was 1 mg·L^-1 after treatment for 24 h. When the concentration was 500 mg·L^-1， water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages， and at this concentration， the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted， the phagocytic index was >113%. In addition， the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS， the inhibitory rate was >45%. Conclusion:Combined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang， this study establishes an <italic>in vitro </italic>biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages， and 500 mg·L^-1 was confirmed as the limit concentration. Under the limit concentration， Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS， which can be judged as qualified.

Screening of Qualitative Preparation Quality Markers of Yuliantang / 中国实验方剂学杂志

Objective:To screen qualitative preparation quality markers of Yuliantang， in order to provide data support for the selection of indicator components， and establish the direct connection between indicator components and efficacy （Xiehuo Zhitong） for achieving the quantity-effect combination. Method:The stability of preparation process of Yuliantang lyophilized powder was investigated by HPLC fingerprint technology， then， the components in Yuliantang lyophilized powder were identified by UHPLC-LTQ-Orbitrap-MS. By referring to the relevant literature， the pharmacological activities of these identified compounds were compared with the pharmacological effects corresponding to the efficacy of Yuliantang， and the composition of the qualitative preparation quality markers of Yuliantang lyophilized powder was determined. Result:The similarities between HPLC fingerprint of 10 batches of Yuliantang lyophilized powder and the control fingerprint were >0.9， indicating that the preparation process was stable and feasible. A total of 29 components were identified from Yuliantang， of which 23 alkaloids， 3 phenylpropanoids， 2 sesquiterpenoids and 1 limonoid， and there were 15 ingredients of<italic> </italic>Coptidis Rhizoma， 12 ingredients of<italic> </italic>Euodiae Fructus， and 2 ingredients of<italic> </italic>Aucklandiae Radix. The composition of the qualitative preparation quality markers of Yuliantang was initially determined as magnoflorine or 10-hydroxy-2，3，9-trimethoxyberberine， phellodendrine， menisperine， thalifendine， groenlandicine， dehydroevodiamine， coptisine， jatrorrhizine， columbamine， methylcoptisine， berberine， epiberberine， palmatine， evodiamine， rutaecarpine， neochlorogenic acid， cryptochlorogenic acid， chlorogenic acid， limonin， costunolide， dehydrocostus lactone. Conclusion:The method for researching and screening the preparation quality markers in Yuliantang lyophilized powder is scientific， reasonable and feasible， it can provide reference for the determination of component indicators in the process research of Yuliantang and qualitative and quantitative indexes in its quality standard.

Screening of quantitative preparation quality markers of Yulian Decoction / 中国中药杂志

On the basis of the qualitative preparation quality markers of Yulian Decoction, we screened out the quantitative markers and explored a general strategy for analyzing the component migration in Chinese herbal pieces, preparations, and plasma. A method capable of simultaneously determining 28 chemical components in Yulian Decoction was established based on HPLC-MS/MS. This method was used to determine the migrated components in herbal pieces-lyophilized powder preparations-rat plasma after administration of Yulian Decoction. Liquid chromatography was performed under the following conditions: C_(18)-reversed phase chromatographic column(2.1 mm × 100 mm, 1.8 μm); acetonitrile-water(containing 0.1% formic acid) as the mobile phase for gradient elution; the flow rate of 0.2 mL·min~(-1). Electrospray ionization source was adopted for mass spectrometry detection, in which positive and negative ion modes and multiple reaction monitoring were applied. Confirmed by the methodological investigation in linear range, recovery(95.48%-103.4%), precision(RSD, 0.45%-3.8%), stability, and repeatability(RSD, 5.6%-14%), the established method was suitable for the detection and quantification of the components in Yulian Decoction. The results showed that in the lyophilized powder of Yulian Decoction, berberine was greater than 5% in mass fraction, magnoflorine, epiberberine, coptisine, palmatine, and limonin in the range of 1%-5%, and dehydroevodiamine, evodiamine, rutaecarpine, costunolide, and dehydrocostus lactone in the range of 0.002%-1%. Of the 28 components detected in pieces, 27 were found to migrate to the lyophilized powder, and 11 were detected in rat plasma. Fifteen components were preliminarily determined as quantitative preparation quality markers for Yulian Decoction, including berberine, epiberberine, coptisine, palmatine, evodiamine, rutaecarpine, limonin, costunolide, dehydrocostus lactone, magnoflorine, jatrorrhizine, columbamine, groenlandicine, chlorogenic acid, and neochlorogenic acid. In conclusion, the HPLC-MS/MS general strategy was established for analyzing the migration of multiple components in Chinese herbal pieces, preparations, and plasma, which can provide the basis for the screening of quantitative preparation quality markers and multi-index quality control of Yulian Decoction.

Preparation and in vitro evaluation of puerarin nanoemulsion lyophilized powder with glycyrrhizic acid as stabilizer / 中草药

Objective: Puerarin nanoemulsion lyophilized powder (Pue-NE-LP) was prepared using natural surfactant glycyrrhizic acid as stabilizer and evaluated in vitro. Methods: Pue-NE was prepared by high-speed shear and high-pressure homogenization method, and further combined with freeze-drying method to prepare Pue-NE-LP. Taking the average particle size and polydispersity index (PDI) as the evaluation indexes, the optimal prescription and process parameters of this experiment were screened out through a single factor test. The prepared Pue-NE-LP was characterized by physicochemical properties and dissolution in vitro. Results: The average particle size and PDI of Pue-NE-LP prepared with 5% glyceryl caprylate as oil phase, 2.0 mg/mL glycyrrhizic acid as stabilizer, and 7% glucose as lyophilization protectant was (215.1 ± 0.7) nm and (0.133 ± 0.024), respectively. Scanning electron microscopy showed that Pue-NE-LP was irregularly small and uniform in size; X-ray diffraction showed that Pue-NE-LP existed in an amorphous state. In vitro release results showed that the dissolution rate of Pue-NE-LP was significantly higher than the physical mixture. Conclusion: Pue-NE-LP prepared with natural surfactant glycyrrhizic acid as a stabilizer is not only simple to prepare, but also can significantly improve the solubility and bioavailability of puerarin. It provides a reference for the multiple development of Pue-NE formulations.

Content Determination of 4 Indicator Components in Shengyu Decoction Lyophilized Powder by HPLC / 中国药房

OBJECTIVE：To establish the content determin ation method of ferulic acid ，verbascoside，ligustilide and astragaloside in Shengyu decoction lyophilized powder. METHODS ：HPLC method was adopted to determine 4 components in 3 batches of lyophilized powder. The determination of ferulic acid ，verbascoside and ligustilide was performed on Inertsil ODS-SP C 18 column with mobile phase consisted of methanol- 0.1％ phosphoric acid （gradient elution ）at the flow rate of 1.0 mL/min；detector was diode array detector ；detection wavelength was set at 330 nm；column temperature was 30 ℃，the sample size was 10 μL. The determination of astragaloside was performed on Kromasil C 18 column with mobile phase consisted of acetonitrile-water （32∶68，V/ V）；detector was evaporative light scattering detector ；the drift tube temperature wa s 100 ℃，the carrier gas （air）flow rate was 2.5 L/min at the flow rate of 1.0 mL/min；column temperature was 30 ℃，the sample size was 10 μL. RESULTS：The linear ranges of ferulic acid ，verbascoside，ligustilide and astragaloside were 0.050 15-10.03 μg（r＝0.999 8），0.067 80-13.56 μg（r＝ 0.999 9），0.057 30-11.46 μg（r＝0.999 5），1.128-11.28 μg（r＝0.999 3），respectively. The detection limits were 2.12×10－4，1.30× 10－3，8.02×10－4，1.09×10－3 μg，respectively. The limit of quantification were 7.43×10－4，3.87×10－3，2.34×10－3，3.36×10－3 μg， respectively. RSDs of precision ，stability（12 h）and reproducibility tests were all lower than 2％（n＝6）. Average recovery rates were 99.6％（RSD＝0.83％，n＝6），100.9％（RSD＝1.07％，n＝6），98.8％（RSD＝0.84％，n＝6）and 101.3％（RSD＝0.99％， n＝6），respectively. The contents of ferulic acid ，verbascoside，ligustilide and astragaloside in 3 batches of samples were 1.225-1.248， 0.413-0.424， 0.325-0.332， 0.394-0.404 mg/g， respectively （RSDs among batches were lower than 1.5％ ）. CONCLUSIONS：Established method is stable ，reproducible，rapid and accurate for the content determination of ferulic acid ， verbascoside， ligustilide and astragaloside in Shengyu

Preparation of Oxymatrine Phospholipid Complex Solid Lipid Nanoparticles Lyophilized Powder and Evaluation of Its Quality / 中国实验方剂学杂志

Objective: To prepare oxymatrine phospholipid complex solid lipid nanoparticles(OMT-PC-SLN) lyophilized powder and evaluate its pharmaceutical properties. Method: Pseudo-ternary phase diagram was employed to optimize the formula of microemulsion;single factor experiments were adopted to optimize the formulation process of OMT-PC-SLN lyophilized powder with encapsulation efficiency as index;the morphology of this preparation was observed by transmission electron microscope(TEM).The particle size was measured by particle size analyzer and the in vitro release performance of OMT-PC-SLN lyophilized powder was examined. Result: Optimal formulation process was as following:taking soybean phospholipid and polyethylene glycol 15-hydroxystearate(Kolliphor HS 15) as the emulsifier,ethanol as co-emulsifier,ratio of emulsifier to co-emulsifier(Km)=3:2,oil phase:(emulsifier+co-emulsifier)=1:9,oxymatrine phospholipid complex-stearic acid-soybean phospholipid-Kolliphor HS 15-ethanol(30:100:180:360:360);taking 50 mL of 4%mannitol solution as the external aqueous phase,ice bath stirring at 1 000 r·min-1 and solidifying for 1 h,precooled at -20℃ for 24 h,took out and dried for 24 h.OMT-PC-SLN lyophilized powder was spherical in appearance with encapsulation efficiency of (38.09±1.24)%,average particle size of 785.5 nm,polydispersity coefficient(PDI) of 0.456 and the Zeta potential of -24.82 mV.The cumulative release rates of OMT-PC-SLN lyophilized powder were 72.63%at 2 h and 98.42%at 12 h;the cumulative release rate of oxymatrine(crude drug) was 98.60%at 2 h. Conclusion: This optimized formulation process of OMT-PC-SLN lyophilized powder is stable with good repeatability;compared with oxymatrine,OMT-PC-SLN lyophilized powder has a certain sustained-release effect.

Preparation of sustained-release tablets of tilianin nanosuspension lyophilized powder / 中草药

Objective: To prepare sustained-release tablets of tilianin nanosuspension lyophilized powder. The factors that might influence drug release and release mechanism were studied in present study. Methods: High pressure homogenization method was used to prepare tilianin nanosuspension. Lactose and mannitol (3:1) were employed as freeze-drying protective agent to prepare lyophilized powder. HPMC was used as framework material to prepare sustained-release tablets of tilianin nanosuspension lyophilized powder. Based on single factor test, the effects of proportion and amounts of HPMC K4M and HPMC K15, amounts of PEG 4000 and magnesium stearate on in vitro drug release of sustained-release tablets were investigated. Orthogonal test was designed to gain the optimum prescription. Results: The particle size and zeta potential of tilianin nanosuspension were (164.41 ± 9.72) nm and (-37.21 ± 2.38) mV, respectively. The particle size and zeta potential of re-dispersed freeze-drying products were (211.83 ± 11.26) nm and (-31.66 ± 2.92) mV, respectively. The optimum prescription was as follow: the proportion and amounts of HPMC K4M and HPMC K15 were 2:1 and 40 mg, amounts of PEG 4000 was 20 mg, and amounts of magnesium stearate were 0.5%. Sustained release tablets of tilianin nanosuspension were well accorded with Higuchi kinetics model. The equation was Mt/M∞ = 0.286 8 t1/2-0.073 8, r2 = 0.981 4. And the cumulative release could achieve 92.36% in 12 h. The drug release from the tablets was controlled by diffusion and degradation of the matrix. Conclusion: The preparation technology of sustained release tablets of tilianin nanosuspension lyophilized powder has good reproducibility. This sustained release tablets could control the release of tilianin

Preparation and Dissolution of Loratadine Nanosuspension Lyophilized Powder / 中国药师

Objective:To prepare loratadine nanosuspension lyophilized powder (LTD-NLP) and investigate the dissolution and stability. Methods:LTD-NLP was prepared by an anti-solvent precipitation method; the particle size and morphology were characterized by a laser nanometer particle sizer and a transmission electron microscope (TEM). The optimal lyophilized protective agent was screened out; the dissolution and stability of LTD-NLP were determined by HPLC. Results:The average particle size of LTD-NLP was 175.6 nm with PDI of 0.200, and the zeta potential was -56.3 mV. The shape of LTD-NLP was spherical. The nanosuspension lyophilized with 10% sucrose presented optimal properties. The dissolution of LTD-NLP was greater than raw material, and LTD-NLP had good stability at 4℃. Conclusion:The anti-solvent precipitation method for LTD-NLP is simple,and the product is expected to become a new preparation of loratadine.

Preparation, characterization, and dissolution characteristics of ursolic acid nanoparticles prepared by emulsion solvent evaporation method / 中草药

Objective To prepare and characterize ursolic acid nanoparticles (UANs), and to investigate its improvement of equilibrium solubility and dissolution rate. Methods UANs were prepared by emulsion solvent evaporation method, and followed by freeze-drying. The organic phase was trichloromethane containing 30% ethanol, the aqueous phase is ultrapure water, poloxamer 188 was as surfactant and cryoprotectant. The optimal conditions for preparing nanoparticles were screened out using single-factor experiment. The particle size was used as the basis for optimization experiment. The following six main parameters had significant influences on particle size were picked out, including the concentration of poloxamer 188, volume ratio of organic to water phase, homogenate speed and homogenate time, as well as homogenization pressure and cycles. And then, dynamic light scattering equipment was used to analyze the mean particle size, the morphology of UANs powder obtained was presented by scan electronic micro-scope (SEM). The UANs weather and how changes in surface chemical character and physical structure was estimated by using X-ray diffraction (XRD) and differential scanning calorimetry (DSC). The equilibrium solubility study and dissolution test were carried on raw ursolic acid (UA) and UANs. Results The optimal conditions of preparation UANs: 0.05% of poloxamer 188, 1:4 of volume ratio of organic to water phase, 7 000 r/min of homogenate speed for 2 min and a homogenization pressure of 50.0 MPa for 6 times. Based on the optimal conditions, the mean particle size was (157.5 ± 28.0) nm and Zeta potential of (20.33 ± 1.67) mV. Particle distribution of UANs illustrated that UA had been nanoscale with uniform particle size distribution. SEM showed that UANs were nearly spherical. By the XRD and DSC, we could acquaint UA in UANs had the same chemical structure as the raw UA but had been amorphous state. The result of solubility test figured that the equilibrium solubility of UANs was 13.48 times in SGF, 11.79 times in SIF and 23.99 times in deionized water than raw UA. The dissolution rate of UANs prepared by ESE method has been up to 14.72 times in SGF and 74.35 times in SIF. Conclusion This study indicates that the emulsion solvent evaporation method has an array of valued on improving water solubility of UA, and it will have benefit on enhancing oral bioavailability.

Clinical effect of Spleen Aminopeptide Oral Lyophilized Powder combined with azithromycin on mycoplasma pneumonia in children and its influence on immune function / 药物评价研究

Objective To investigate the clinical effect of Spleen Aminopeptide Oral Lyophilized Powder on mycoplasma pneumonia in children and its influence in immune function.Methods Totally 103 cases of children infected with mycoplasma pneumonia from May 2013 to June 2016 were selected and divided into observation group and control group,with 53 cases in each group.The control group were treated with azithromycin,and observation group were treated with azithromycin combined with Spleen Aminopeptide Oral Lyophilized Powder.The clinical effect,T cell subgroup (CD4+,CD8+,CD4+/CD8+) levels,serum cytokine levels (IL-10,IL-17,TGF-β1) and safty were compared.Results The total effective rate of the observation group was 94.39％,which was significantly higher than that of the control group of 79.25％ (P ＜ 0.05).After treatment,the levels of CD4+,CD4+/CD8+ in two groups were significantly higher than those before treatment (P ＜ 0.05),and the levels of CD4+,CD4+/CD8+ in observation were higher than those in control group significantly (P ＜ 0.05).The levels of CD8+,IL-10,IL-17,TGF-β1 in two groups were significantly lower than before treatment (P ＜ 0.05),and those indexes in observation were lower than those in control group,the difference was statistically significant (P ＜ 0.05).Conclusion Azithromycin combined with Spleen Aminopeptide Oral Lyophilized Powder has remarkable clinical effect on mycoplasma pneumonia in children,which is cecurity and reliable,and can improve the cellular immune function and reduce the immune injury.

Clinical effect of Spleen Aminopeptide Oral Lyophilized Powder combined with azithromycin on mycoplasma pneumonia in children and its influence on immune function / 药物评价研究

Objective To investigate the clinical effect of Spleen Aminopeptide Oral Lyophilized Powder on mycoplasma pneumonia in children and its influence in immune function.Methods Totally 103 cases of children infected with mycoplasma pneumonia from May 2013 to June 2016 were selected and divided into observation group and control group,with 53 cases in each group.The control group were treated with azithromycin,and observation group were treated with azithromycin combined with Spleen Aminopeptide Oral Lyophilized Powder.The clinical effect,T cell subgroup (CD4+,CD8+,CD4+/CD8+) levels,serum cytokine levels (IL-10,IL-17,TGF-β1) and safty were compared.Results The total effective rate of the observation group was 94.39％,which was significantly higher than that of the control group of 79.25％ (P ＜ 0.05).After treatment,the levels of CD4+,CD4+/CD8+ in two groups were significantly higher than those before treatment (P ＜ 0.05),and the levels of CD4+,CD4+/CD8+ in observation were higher than those in control group significantly (P ＜ 0.05).The levels of CD8+,IL-10,IL-17,TGF-β1 in two groups were significantly lower than before treatment (P ＜ 0.05),and those indexes in observation were lower than those in control group,the difference was statistically significant (P ＜ 0.05).Conclusion Azithromycin combined with Spleen Aminopeptide Oral Lyophilized Powder has remarkable clinical effect on mycoplasma pneumonia in children,which is cecurity and reliable,and can improve the cellular immune function and reduce the immune injury.

The influence of spleen aminopeptidase lyophilized powder on serum levels of TGF -β1,MCP -1,SDF -1 in pediatric asthma / 中国基层医药

Objective To investigate the influence of spleen aminopeptidase lyophilized powder on serum levels of transforming growth factor -β1 (TGF -β1),monocyte chemoattractant protein -1 (MCP -1),stromal cell derived factor 1 (SDF -1 )in pediatric asthma.Methods 128 patients with asthma were randomly divided into observation group(64 cases)and control group (64 cases).Two groups of children were given symptomatic treat-ment.The control group was orally administered montelukast,the observation group was treated with montelukast and splenic ammonia lyophilized powder for 3 months.The levels of TGF -β1,MCP -1,SDF -1 of the two groups were compared before and after treatment.Results After treatment,the asthma control test table (ACT)score,expiratoryvolume in one second (FEV1 )and peak expiratory flow rate (PEF)in the observation group were (24.25 ± 3.98)points (80.25 ±4.25)%,(7.25 ±0.69)L/min,which were significantly higher than those in the control group [(20.12 ±4.02)points,(75.02 ±3.96)%,(5.82 ±0.70)L/min,t =7.203,6.757,14.459,26.677,P <0.01].After treatment,the serum levels of TGF -β1,MCP -1,SDF -1 of the observation group were (42.23 ± 6.02)ng/mL,(48.56 ±3.96)pg/mL,(252.36 ±32.22)ng/L,which were significantly lower than those in the con-trol group [(42.23 ±6.02)ng/mL,(59.02 ±4.22)pg/mL,(425.25 ±40.62)ng/L,t =6.757,14.459,26.677,all P <0.01].Conclusion Montelukast combined with splenic aminopeptidase lyophilized powder for asthma can effec-tively reduce serum TGF -β1,MCP -1,SDF -1 levels and improve clinical symptoms and lung function,improve the treatment of children.

Preparation, optimization, and dissolution characteristics of curcumin nanoparticles lyophilized powder by antisolvent method / 中草药

Objective: Curcumin nanoparticles lyophilized powder (CNLP) were prepared by antisolvent method which was optimized using single factor method. In this process, acetone was used as solvent, deionized water was used as antisolvent, tween-80 was used as surfactant and mannitol was used as lyoprotectant. Methods: The main factors affecting the particle size of CNLP included the concentration of curcumin, volume ratio of solvent and antisolvent, dosage of surfactant, precipitation time, stirring speed, and dosage of lyoprotectant. The contrast experiments on dissolution in vitro was done between CNLP and raw curcumin powder. Results: The mean particle size of CNLP was (172.2 ± 4.6) nm; The Zeta potential of CNLP redissolving in water was (-19.7 ± 3.7) mV. The SEM graphs indicated the raw curcumin was in irregular and massive shape and its particle size was about 20 μm. The CNLP exhibited regular block structure and its particle size was about 170 nm which was obviously reduced compared with raw curcumin. The mean particle size of CNLP obtained from laser particle analyzer was in accord with the morphology of CNLP. The saturated solubility of CNLP was 41.32 times of raw curcumin powder in deionized water, 1.74 times in simulated gastric fluid and 4.11 times in simulated intestinal fluid through saturated solubility test, respectively. The dissolution rate of CNLP was 14.51 times of raw curcumin powder in water dissolution medium, 2.33 times in simulated gastric fluid and 44.79 times in simulated intestinal fluid through dissolution determination, respectively. Conclusion: The preparation process of CNLP using antisolvent method could improve the drawback of poor water solubility and enhance the bioavailability of curcumin.

Neuroprotective effect of lyophilized powder of catalpol and puerarin on oxygen-glucose deprivation/reperfusion injury in astrocytes of rat cerebral cortex in vitro / 中国药学杂志

OBJECTIVE: To investigate the protection of lyophilized powder of catalpol and puerarin (C-P) on oxygen-glucose deprivation/reperfusion(OGD/R)-injured astrocytes and the possible mechanism in vitro. METHODS: Primary astrocytes were isolated from the cerebral cortex of neonatal rats. The purity of astrocytes was identified by GFAP immunofluorescence. Astrocytes were divided into 6 groups: normal group, model group, excipients group, and three groups with gradient doses of C-P (12.25, 24.50, 49.00 μg·mL-1). After astrocytes suffered from OGD/R (OGD 6 h/R 12 h) with or without C-P, cell survival, LDH leakage, and apoptosis were analyzed by MTT, colorimetry, and TUNEL respectively. The apoptotic protein, caspase-3, was evaluated by Western blot. Meanwhile, oxidative indexes including SOD, GSH, ROS, MDA, and NO were detected by assay kits. In terms of inflammation, TNF-α, IL-1β, and PGE2 in medium were evaluated via ELISA. iNOS, COX-2, NF-κB p65, p-NF-κB p65, IκB-α, and p-IκB-α were examined by Western blot. RESULTS: The purity of primary astrocytes was more than 97%. Compared with normal group, the cell survival rate of model group decreased significantly, while the LDH leakage rate and cell apoptosis rate markedly increased after OGD/R injury in model group. Meanwhile, SOD activity and GSH level in astrocytes markedly decreased. The level of MDA and ROS significantly increased. The content of NO, TNF-α, IL-1β, and PGE2 released in medium also sharply increased. However, those changes of related biochemical indexes above could be reversed by different gradient doses of C-P. There was no significant difference between excipients group and model group. Further study found that C-P could inhibit the protein expression of caspase-3, iNOS, and COX-2, as well as the increase of phosphorylation level of NF-κB p65 and IκB-α significantly. CONCLUSION: C-P shows a protective effect on the OGD/R injured astrocytes in vitro, and its protection mechanism involves in anti-inflammation, antioxidant, inhibiting the cell apoptosis and activation of NF-κB signaling pathway.

Preparation and in vitro release of lyophilized powder of baicalein solid lipid nanoparticles / 中草药

Objective: To prepare lyophilized powder of baicalein solid lipid nanoparticles (BA-SLNs) and investigate its physicochemical properties and release characteristics. Methods: Ba-loaded SLN was prepared by solvent emulsification-evaporation method, the formulation was optimized by orthogonal design, with encapsulation efficiency (EE) as reference, the measurement of particle size, morphology, Zeta potential, the polydispersity index (PDI), and in vitro drug release behavior. The lyophilized powder of appearance, color, redispersibility as indexes, the differential scanning calorimetry (DSC), X-Ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) were used to analyze its material phase of the drug in nanoparticles. Results: The Ba-SLNs assumed a spherical shape with an even distribution of diameter and particle size was (82.64 ± 6.78) nm, the PDI was 0.242 ± 0.013, Zeta potential was (-25.7 ± 0.5) mV, EE was (81.3 ± 1.2)%, and drug loading was (7.16 ± 0.14)% (n = 3). The 5% mannitol was the best protective agent for lyophilized powder of Ba-SLNs. Through the characterization indicated that the drug to amorphous state dispersed in a lipid. In vitro dissolution experiments showed Ba-SLNs compared with pure drugs had obviously sustained release. Conclusion: The technique of preparing Ba-SLN by solvent emulsification-evaporation has small particle size, high EE, and good stability, and the process is simple.

Study on the Quality Standard of Fresh Gastrodia elata Lyophilized Powder / 中国药房

OBJECTIVE:To provide reference for the establishment of quality standard for Fresh Gastrodia elata lyophilized powder. METHODS:TLC was conducted for the qualitative identification of G. elata in preparation,and HPLC was conducted for the content determination of gastrodin in preparation. The column was Diamonsil C18 with mobile phase of acetonitrile-0.05% phos-phoric acid(3∶97,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 25 ℃ and vol-ume injection was 10 μl. RESULTS:TLC pots of Fresh G. elata lyophilized powder were clear and well-separated. The linear of gastrodin was 2.9-14.5 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.00%;recov-ery was 98.07%-102.70%(RSD=1.74%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible,and suitable for the quality control of Fresh G. elata lyophilized powder.

Effect of Spleen Aminopeptide Oral Lyophilized Powder on urinary RBP and CRP in children with mycoplasma pneumoniae infection and asthma / 中国生化药物杂志

Objective To investigate effect of Splenic Aminopeptide Oral Lyophilized Powder on urinary retinol-binding protein and CRP in children of mycoplasma pneumonia infection with asthma .Methods 60 cases were randomly divided into 2 groups, 30 cases of control group were given conventional anti-inflammatory treatment, azithromycin.30 patients of Experimental group were treated with Spleen Aminopeptide Oral Lyophilized Powder treatment, two groups were treated for 7 days.30 cases of healthy children were selected, given no any treatment.After treatment, urinary retinol binding protein, blood urea nitrogen, creatinine, cough, wheezing symptom recovery, C-reactive protein, blood and other changes were compared between two groups.ResuIts Two groups of children with urinary retinol binding protein, urea nitrogen, creatinine levels were improved in different degrees, and in experimental group, urinary retinol binding protein, urea nitrogen and creatinine decreased significantly compared with control group (P<0.05).Two groups of children with asthma and wheeze improved in different degrees, and cough wheeze, cough and wheeze recovery time of experimental group had more improvement than those of control group ( P<0.05 ) .Two group of C reactive protein and white blood cell level both improved in different degrees, and the experimental group of C reactive protein and white blood cell levels were significantly lower than those of control group (P<0.05).ConcIusion Spleen Aminopeptide Oral Lyophilized Powder can significantly improve the infection status of children with mycoplasma pneumonia, and relieve cough and wheeze symptoms, lower urinary retinol binding protein and C reactive protein level, improve kidney function damage induced by hypoxia.

Preparation of lyophilized powder in oral Pueraria flavonoids loaded solid lipid nanoparticles and determination of release rate of its four effective components / 中草药

Objective: To prepare the lyophilized powder of Pueraria flavonoids loaded solid lipid nanoparticles (PF-SLN) and determine the dissolution rate of its four effective components: 3'-hydroxypuerarin, puerarin, daidzin, and daidzein. Methods: PF-SLN was prepared by the high pressure homogenization (HPH) technology. The lyophilized formula contained mannitol as cryoprotectant. The release rates of the four effective components from the PF-SLN lyophilized powder as well as the physical mixture were determined, with artificial gastric juice (pH 1.2) as dissolvent. Results: The technical parameters of PF-SLN preparation optimized by orthogonal test were as follows: The ratio and the dosage of lipid-surfactant were 2:1 and 2.0%, PF dosage was 2.5%, and 150 MPa homogeneity was 15 cycles. The optimal PF-SLN lyophilized powder was loosen with the particle size of (517.1 ± 10.3) nm, polydisperse index of 0.484 ± 0.210, and Zeta potential of (-21.91 ± 2.03) mV, respectively. The in vitro accumulated dissolution rates of PF-SLN lyophilized powder were slower than those of the physical mixture. Conclusion: The method employed to prepare PF-SLN lyophilized powder is feasible. PF-SLN lyophilized powder could delay the in vitro dissolution rate notablely. It might be a novel vehicle potentially for nano-drug delivery system of Pueraria flavonoids.