HPLC-UV/VIS for Determination of Ipratropium Bromide Mixed with Salbuterol, Beclomethasone Propionate and Budesonide using Dual Wavelength-Detection Method.

BACKGROUND: Inhalation preparation involves liquid or solid raw materials for delivering to lungs as aerosol or vapor. The liquid preparation for nebulizer is effective for convenient use and patient compliance and it has been extensively used in the treatment of clinical lung diseases. Clinical staff often mixes the compound ipratropium bromide with beclomethasone propionate and budesonide inhaler but reference values of inhalants for clinical use need to be established for simplifying the operation procedure. The high-performance liquid chromatography (HPLC) method of compound ipratropium bromide solution, beclomethasone propionate suspension and budesonide suspension after mixed atomization was studied. METHODS: The specificity, linearity, recovery (accuracy), precision and stability of compound ipratropium bromide, beclomethasone propionate and budesonide were tested to verify the developed liquid phase method. RESULTS: The developed liquid phase method had high specificity, linear R2≥0,999, recovery (accuracy) RSD (relative standard deviation) less than 2%, precision RSD less than 2,0%, and stability RSD less than 2,0%. CONCLUSION: The liquid phase methodology developed in this study can be used for the determination of compound ipratropium bromide mixed with beclomethasone propionate and budesonide. The current methodology can also be used to provide a reference for the determination of its content after mixing, and further data support for its clinical medication.

Comprehensive chemical profiling and quantitative analysis of ethnicYi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS / 中国天然药物

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

Preparation and identification of specific chicken egg yolk immunoglobulins against cell wall protein of Trichophyton mentagrophytes / 中国免疫学杂志

Objective:Prepared the specific chicken egg yolk immunoglobulins (IgY) against the cell wall protein of Trichophyton mentagrophytes (tmCWP) and detected its biological activities,which was to establish the basis for the preventment and treatment in dermatophytes disease.Methods: In this work,tmCWP was extracted and purified by cold alkali method,and being used as immunogen to immunized healthy laying hens.The IgY was extracted from the egg yolk by polyethylene glycol method and purified by saturated ammonium sulfate method,respectively.The concentration of the extracted IgY was detected by Bradford method.The purity and molecular weight of the specific anti-tmCWP IgY were analysed by SDS-PAGE.The titer of IgY was obtained by ELISA.The immunoreactivity of IgY was performed by Western blot.Results: The purity of the extracted IgY reached to 87.27％.ELISA indicated that the titer of the specific anti-tmCWP IgY gradual rised 20 days after primary immunization and reached to the highest value (1∶32 000) after 45 days.Western blot revealed that the specific IgY showed a good immunoreactivity and a specifically combination capacity.Conclusion: In our work,the tmCWP could be regarded as the immunogen to prepare the specific anti-tmCWP IgY,which could provide a novel thought for the therapy of Trichophyton mentagrophytes infection.

Construction of an eukaryotic expression vector encoding human granzyme B and it's expression in Hep2 cells / 中国病理生理杂志

AIM: To construct pVAX1-GrB. METHODS: Lymphocytes from human laryngeal carcinoma tissue were separated from tumor tissue. The fragment of granzyme B (GrB) was amplified by RT-PCR and was recombined to the downstream of T7 promoter in the vector pVAX1. The construction was transfected into Hep2 cells with lipofectamine 2000. The expression of protein was identified by indirect immunofluorescent antibody assay. RESULTS: It has been proved that the sequence of the RT-PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The GrB cDNA fragment was cloned into the vector of pVAX1 in the right direction and the open reading fragment of GrB was maintained. The target protein was detected in the transfected Hep2 cells. CONCLUSION: The pVAX1-GrB plasmid was successfully constructed and expressed. [