Study On The Quality Standard Of Freeze-drying Process Of Fresh Radix Rehmanniae?Quality Standard Of Freeze Dried Radix Rehmanniae And Preliminary Screening Of The Effective Part Of Its Hypoglycemic Agents

Objective: Vacuum freeze drying conditions of fresh Rehmanniae, establish the vacuum freeze drying process and quality evaluation of the hypoglycemic effective position of preliminary screening of freeze dried Rehmanniae. The main contents of this paper are divided into three parts as follows:(1) the parameters of vacuum freeze drying of fresh rehmannia root to make a systematic investigation, determine the technology of vacuum freeze drying.(2) with a macroscopic, microscopic and physicochemical method to establish the freeze dry method for identification of Radix Rehmanniae; water shall be made in accordance with the relevant methods of Chinese Pharmacopoeia 2010 Edition appendix, as well as ash, heavy metals, arsenic salt, pesticide residues of examination; by HPLC fingerprint to identify and control the quality of freeze dried and raw rehmannia root; determinate the content of trace elements, Catalpol, acteoside and develop freeze-dried criteria of quality evaluation of freeze dried Rehmanniae.(3)Frozen dry extracts from different parts of Radix Rehmanniae on alpha gluco glycosidase inhibitory effect and to reduce its preliminary screening of effective fraction of in vitro sugar; frozen dry extracts from different parts of Radix Rehmanniae on alloxan induced diabetic mice blood glucose and blood lipid and the in vivo reduced further screening of the effective fraction of sugar.Methods: 1. Take catalpol, verbascoside and water content as the index, through the single factor experiment, the main factors influencing the quality of Radix Rehmanniae in vacuum freeze-drying technology; 2. By microscopic and TLC identification of freeze dried Rehmannia from different batches. 3. Inspect the content of water, total ash, acid insoluble ash of dried Rehmannia from different batches according to the 2010 Edition"CHP” appendix IX; by flame atomic absorption spectrometry and atomic fluorescence spectrometry method for the determination of two different areas of Pb, Cd, Cu, As and Hg contents of 5 kinds of heavy metals and harmful elements, by gas chromatography to detect residues of organochlorine pesticides in freeze dried Rehmannia from different areas;4. By HPLC, generating HPLC, into "traditional Chinese medicine chromatographic fingerprint Effects of diet in different parts of the freeze dried and freeze-dried version of spectral similarity evaluation system "(2004a) software similarity evaluation; 5. Using HPLC for the determination of Catalpol Mullein glucoside content in Rehmannia, by flame atomic absorption method for the determination of different habitats in rehmannia root trace elements K, Mg, CA, Na, Mn, Fe, Cu and Zn content; 6. Using UV visible light spectrophotometry extract on alpha glucosyl glycosidase inhibitory effect of freeze driedrehmannia root. 7. To acarbose as positive control were freeze dried Rehmannia different extracts on alloxan induced diabetic mice, water intake and body weight, blood glucose and blood lipid.Results: 1. The best process for material slicing method for crosscutting: slice thickness for 2.5~4mm thick slices; material thickness is double; pre-freezing conditions are-30?, 2h; drying time is 28 h, vacuum degree is 5pa, the material temperature is from-40 ? to 15 ?, the temperature of cold trap is-80?; vacuum packaging material PV+PE sealing temperature in the temperature, sealing time is 3S, vacuum time 20~30s, vacuum degree for-0.1mpa.2. Under the microscope observation to the secretory cells and cork cells, parenchyma cells, bordered lines guide hole, reticulate vessel; in freeze dried Rehmanniae TLC detected catalpol and verbascoside; freeze dried Rehmanniae root of moisture, ash, acid insoluble ash did not exceed the limits specified;3. The content of moisture, ash, acid insoluble ash, harmful elements and pesticide residue in freeze dried Rehmannia did not exceed the limits specified.4. The HPLC chromatograms by similarity evaluation system established 10 common peaks, the establishment of the freeze dried Rehmannia fingerprint and the fingerprints of10 batches of medicinal materials and the control of fingerprint similarity in 0.977-0.998.The established method is simple and reliable.5. Determination the content of catalpol in freeze dried Rehmannia of the different origins by HPLC, we found the smooth baseline components were well separated. FAAS method was used to determine the trace elements in the frozen dried radix. The accuracy,repeatability and recovery rate of the method were good. The trace elements in different producing areas were different.6 UV methods was used to determine the inhibitory effect of extracts of alpha glucosidase in freeze dried rehmannia root, we find that the n-butanol and water and water extracts showed higher enzyme inhibitory activity.7. Taking acarbose as positive drug, with alloxan modeling, we found that the treatment group oral administered after two weeks can reduce diabetic mouse diet and water intake, weight have varying degrees of increase blood glucose was different degrees of decreased, to varying degrees of drop lower diabetes mice postprandial blood glucose,improve glucose tolerance, increase the content of HDL-C and reduce TG, TC.Conclusion: The vacuum freeze drying process stability experiment of Rehmannia is good; the indicators of freeze-dried products in line with the "CHP" the provisions of the2010 Edition; the determination methods of the content of harmful elements, trace elements, catalpol and verbascoside in freeze-dried rehmanniae are accurate, sensitive and repeatable; the method of freeze-dried Rehmannia fingerprint is simple and reliable, it can be used to formulate the quality and evaluation standard of freeze dried Rehmannia; in vitro and in vivo studies showed that all the various parts have the hypoglycemic effect, In vitro studies showed parts of alpha glucosidase inhibitory rate from high to low is as follows: the alcohol extract matter parts of water, acarbose, Rehmannia polysaccharide,water extractive fractions, ethanol extraction of butanol and alcohol provided material of ethyl acetate, alcohol extracts of petroleum ether fraction; in vivo studies found that ethanol extraction of butanol of freeze dried Rehmannia glutinosa can significantly reduce blood sugar, improve glucose tolerance, and all treatment groups can increase the content of HDL-C, reduce the content of TC and TG.