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Objective To determine the technological conditions for the purification of the total alkaloid from the Corydalis Rhizoma (CR) by macroporous adsorption resin. Methods Total alkaloids of CR were determined by acid dye colorimetry, palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline were determined by HPLC. Six macroporous adsorption resins were investigated with the absorption rates, elution rates, and the content of the total alkaloid and four alkaloids of CR by static and dynamic adsorbing experiments. The purification process conditions of the total alkaloid of CR were optimized by the loading amount and volume flow of sample, the type and volume of the impurity removal of solvent and elution solvent and so on. The stability of the purification process was investigated by 5, 10 times enlargement. Results D141 type macroporous adsorption resin was the best choice for the purification of the total alkaloid from CR, the optimized parameters were as follows: Drug concentration was 0.6 g/mL of medicinal material and was added to the D141 macroporous resin column that the ratio of diameter to height range from 1:5 to 1:9 at a flow rate of 2 BV/h to 2 BV, 1.3 BV of purified water was used to remove impurities, and then 6 BV 95% ethanol was used as eluent at a flow rate of 2 BV/h. The purity of the total alkaloid of CR was up to 68.19% after purification, and the content of palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline was 1.95%, 11.74%, 4.93%, and 6.36%, respectively. The purity of the total alkaloid of the CR can reach more than 65% by 5 times and 10 times enlargement. Conclusion The purity of the total alkaloids can reach more than 65% after verification test, and the transfer rate of total alkaloids and four alkaloid monomers of CR can reach more than 85%, indicating that D141 macroporous adsorption resin can effectively purify total alkaloids from CR, and can be applied to industrial production.
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Objective:To optimize the ultrasonic extraction process of total alkaloids from Oxytropis falcata bunge. Methods:The independent variables were solvents ratio, extracting time and ethanol concentration, and the dependent variable was content of total al-kaloids. Based on single factor tests, central composite design and response surface methodology was adopted to optimize the extraction technology. Results:The optimal extraction conditions were as follows: extracted twice with 36-fold amount of 72% ethanol ( contai-ning 1% acetic acid) at 60 ℃, and extracted 77 minutes each time. Under the above conditions, the content of total alkaloids was 2. 793 mg·g-1 with the bias ratio less than 2% when compared with the model predictions. Conclusion:Ultrasonic extraction process of total alkaloids from Oxytropis falcata Bunge optimized by central composite design and response surface method is simple, highly pre-cise, reliable and predictable.
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Objective To screen endophytic fungal strains isolated from wild Huperzia serrata which can produce alkaloids and huperzine A (HupA). Methods Thirty-three endophytic fungal strains were obtained from H.serrata.Alkaloid production was assayed with alkaloid precipitation and acid dye colorimetry. Then, the crude extracts of alkaloids produced from fungal strains were analyzed by modulation of acetylcholinesterase(AChE) activity, TLC and HPLC. Identification of endophytic fungi producing HupA was based on morphology analysis. Results Seven alkaloids produced by endophytic fungal strains were determined based on alkaloid precipitation and acid dye colorimetry, among which strain V generated HupA in vitro. Morphology analysis showed that strain V belonged to Penicillium. Conclusion A high percentage of fungal strains isolated from H.serrata leaves has alkaloid producing potentials. It is the first report that Penicillium species produced HupA. The screening of endophytic fungi producing HupA based on alkaloid precipitation and acid dye colorimetry, AChE activity and structure identification are more efficient and directive.
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Objective To screen endophytic fungal strains isolated from wild Huperzia serrata which can produce alkaloids and huperzine A (HupA). Methods Thirty-three endophytic fungal strains were obtained from H.serrata.Alkaloid production was assayed with alkaloid precipitation and acid dye colorimetry. Then, the crude extracts of alkaloids produced from fungal strains were analyzed by modulation of acetylcholinesterase(AChE) activity, TLC and HPLC. Identification of endophytic fungi producing HupA was based on morphology analysis. Results Seven alkaloids produced by endophytic fungal strains were determined based on alkaloid precipitation and acid dye colorimetry, among which strain V generated HupA in vitro. Morphology analysis showed that strain V belonged to Penicillium. Conclusion A high percentage of fungal strains isolated from H.serrata leaves has alkaloid producing potentials. It is the first report that Penicillium species produced HupA. The screening of endophytic fungi producing HupA based on alkaloid precipitation and acid dye colorimetry, AChE activity and structure identification are more efficient and directive.
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Objective: To propose the feasibility of the vortex method of acid dye colorimetry replacing extraction method to determine the content of total alkaloids in plasma of rats after oral administration with Mahuang Decoction. Methods: The contents of total alkaloids in plasma of rats orally administered with Mahuang Decoction extracted by traditional extraction method and vortex method were determined by acid dye colorimetry, and the results were contrasted and analyzed. Results: The extraction method and vortex method had good linear relationship in the range of 4-40 and 4-80 μg/mL with the average recovery rates of 95.63% (RSD = 1.92%) and 111.55% (RSD = 0.26%), respectively. The regression equations established were A = 0.0171 C + 0.2355, r = 0.9855 and A = 0.0068 C + 0.0102, r = 0.9964. The metabolic processes of total alkaloids in rats were both complied with the one-compartment model. The main pharmacokinetic parameters AUC, Cmax, tmax, and t1/2 were 5.30 × 105 min·μg/mL, 57.13 μg/mL, 37.20 min, 6403.78 min and 1.94 × 106 min·μg/mL, 663.20 μg/mL, 361.90 min, 1756.87 min. The Cmax and AUC of vortex method were 11.61 times and 3.66 times compared with those of extraction method. Conclusion: The vortex method is more rapid, accurate, sensitive, simple, stable, safe, and reproducible and has little harm to laboratory assistants, which can extract the total alkaloids in maximum, with the necessity, scientificity, and practicality of replacing traditional extraction method.
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OBJECTIVE:To establish a method for content determination of matrine in it's liposomes by acid dye colorime_ try.METHODS:Sephadex gel G-50 column was established by swelling of matrine liposomes sample in distilled water for at least 12 hours,distilled water was used as mobile phase,after eluting,the column was mixed with 0.0 125%of bromothymol bl_ ue buffer solution and chloroform,and then the mixture was shaken,standing and demixed,the absorbability of chloroform layer was detected at the wavelength of 413 nm.RESULTS:The detection concentration of matrine showed good linearity with its absorbability within the range of 0.04～0.20mg/ml(r=0.9 972),the average recovery was 94.10%(RSD=1.86%).CONCL_ USION:The established method is simple and accurate,which can be used for quality control of matrine liposomes.
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OBJECTIVE:To establish a method for content determination of principal agents in kanamycin sulfate eye drops by dye colorimetry.METHODS:On the basis of the spectrometric features of ion pair compound produced in the reaction between kanamycin and bromcresol green in buffer solution with pH value of6.0,absorbance was detected at the wavelength of432nm and content was determined.RESULTS:The detected concentration of Kanamycin showed good linear relationship with its absorbance(r=0.9998)within the range of18.0～72.0?g/ml.The average recovery was99.3%(RSD=1.20%).CONCL_ USION:The established method is rapid,simple and accurate,and it can be used for quick test of this preparation during the production.
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Objective: To explorate the drug releasing rule in vitro of Maxingshigan Dropping pill and Suppository. Methods: Their dissolution rates in vitro were determined by the rotary basket method, the contents of total alkaloid in ephedra alkaloid in the two preparations were determined by acidic dye colorimetry, and they were used as markers of dissolution.Results:The dissolution quantity in 45 min of Dropping pill reached 100%, whereas that of suppository reached 92.58%. Conclusion: T 50 , T d of the two preparations showed significant difference ( P