Study On Quality Evaluation System For Standard Decoction Of Curculigo Orchioides

Objectives:Through the study on the quality evaluation system of the standard decoction of Curculigo orchioides,the quality standard of the standard decoction of Curculigo orchioides was established,which can provide reference for the formulation of quality standard of modern decoction pieces based on the decoction of Curculigo orchioides.Method:1.Orthogonal test was used to study the factors such as softening water consumption,softening time and softening temperature of Curculigo orchioides.The optimal processing technology was determined through the process validation test,and 27 batches of Curculigo orchioides were prepared by this technology.2.The single factor experiment was used to investigate the factors such as the extraction rate,the content of curculigoside and the content of 2,6-dimethoxybenzoic acid.And through the process validation test,the most suitable preparation process was determined,with which 27 batches of decoction of Curculigo orchioides pieces were prepared.In such experiment,we will determine the relative density and pH value,calculate the paste rate of dry paste,and specify the allowable range of paste rate with the mean value±2sd.3.Using high performance liquid chromatography,SymmetryShield^TMM RP18（4.6×250 mm,5μm）column was used;mobile phase:acetonitrile-0.1%phosphoric acid solution（21:79）;detection wavelength:285 nm;flow rate:1.0 mL/min;column temperature:35℃;injection volume:20μL.The content of curculigoside and2,6-dimethoxybenzoic acid in 27 batches of standard decoction of Curculigo orchioides was determined and its transfer rate was calculated.The allowable range of transfer rate was determined by the mean value±2sd.4.HPLC was used to study the fingerprints of 27 batches of decoction of Curculigo orchioides pieces on SymmetryShield^TMRP18（4.6×250 mm,5μm）column;mobile phase:acetonitrile-0.1%phosphoric acid solution,gradient elution;detection wavelength:285 nm;flow rate:0.5 mL/min;column temperature:35℃;injection volume:20μL.5.Through the study on the quality evaluation system of decoction of Curculigo orchioides pieces,the quality standard of decoction of Curculigo orchioides pieces was established,and the pilot scale-up test of three batches of decoction of Curculigo orchioides pieces was carried out to verify the feasibility of the quality standard.Results:1.The processing technology of Curculigo orchioides is as follows:take a proper amount of Curculigo orchioides,divide it into different grades,add 1 time of water to soften it at room temperature（about 20℃）for 9 hours（the soften time of the crude medicine can be appropriately extended to the degree of no hard core）,and stir the medicine once every 1 hour.Take out the softened medicinal materials,cut them into1-1.5 cm sections,and dry them in an oven at a set temperature of 60℃for 6 hours.2.The preparation process of decoction of Curculigo orchioides standard is as follows:take about 100 g of decoction of Curculigo orchioides pieces,add 7 times of water to the first decoction,soak for 30 min,boil with fire,and decoct with fire for 60min;add 5 times of water to the second decoction,boil with fire,and decoct with fire for 40 min.Filter it with 100 mesh sieve while it is hot,cool it,combine the decoction twice,and concentrate it to 500 mL.The 27 batches of standard decoction of Curculigo orchioides were brown clear aqueous solution,which would precipitate after standing.The relative density is 1.009-1.015,the pH value is 5.24-5.56,the average paste yield is 13.09%,and the range of paste yield is 9.63-16.55%.3.The linear equation of curculigoside is y=7927.2x-32.164,R²=0.9999;the linear equation of 2,6-dimethoxybenzoic acid is y=11712x-29.538,R²=1.0000.The average recoveries of curculigoside and 2,6-dimethoxybenzoic acid were 100.94%and 98.73%,respectively.The samples were stable within 24 hours.The RSD of curculigoside was 3.98%,the RSD of 2,6-dimethoxybenzoic acid was 3.87%,the RSD of curculigoside was 0.22%,the RSD of 2,6-dimethoxybenzoic acid was0.77%.The average transfer rate of 27 batches of decoction of Curculigo orchioides pieces was 61.41%based on curculigoside,ranging from 34.97%to 87.85%,and90.42%based on 2,6-dimethoxybenzoic acid,ranging from 70.62%to 100%.4.The fingerprints of 27 batches of decoction of Curculigo orchioides pieces were established and 13 common peaks were matched.Three components of curculigoside,2,6-dimethoxybenzoic acid and chlorogenic acid were identified by comparison with the reference samples.The similarity of 27 batches of decoction of Curculigo orchioides pieces was more than 0.8.5.The standard decoction prepared by the pilot scale-up test is brown clear aqueous solution,which will precipitate after standing.The relative density was 1.010,1.010,1.011,the pH value was 5.44,5.32,5.35,the extract rate was 9.65%,11.81%,11.69%,the transfer rate was 62.29%,80.17%,48.70%respectively,the transfer rate was 81.06%,73.31%,77.98%respectively,and the similarity between the fingerprint and the control fingerprint of 27 batches of samples was above 0.95.Conclusion:In this paper,the relative density,pH value,extract rate and index component transfer rate of the standard decoction of Curculigo orchioides were studied,and the quality standard of the standard decoction of Curculigo orchioides was established by quantitative analysis of index component and qualitative analysis of fingerprint.Through the pilot scale-up test,it is verified that the preparation process of the standard decoction of Curculigo orchioides pieces is stable and feasible.And such process possesses good repeatability,reliable quality,and it has a certain degree of standardization.It can be used as an intermediate reference for the standardization of different drug forms of Curculigo orchioides,and can provide reference for the study of the quality standard of the standard decoction of other medicinal materials.