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OBJECTIVE To optimize extraction technology of couplet medicinals of Astragalus membranaceus-Puerariae lobatae. METHODS With contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract as index，the analytic hierarchy method was used to determine the weight coefficient of each index and calculate the comprehensive score. The effects of solid-liquid ratio， extraction times and extraction time on the comprehensive score were investigated by single factor test. The level of each factor was determined. By multi-index comprehensive scoring method， using comprehensive scores of above 7 indexes as indexes，the extraction technology of couplet medicinals of A. membranaceus-P. lobata was optimized by orthogonal experiment，and the validation tests were conducted. RESULTS The weight coefficient for the contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract were respectively 0.304 7，0.065 2，0.185 8，0.185 8，0.107 8，0.107 8 and 0.042 7. The optimal extraction technology was determined as follows： solid-liquid of 1∶8（g/mL），extracting 3 times and for 1 h each time. RSD of each evaluation index in the validation test results was lower than 3.00% （n＝3）. CONCLUSIONS The optimized extraction technology for A. membranaceus-P. lobata is stable and feasible.
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OBJECTIVE To optimize the extraction technology of modified Tabusen- 2（MT-2），and to investigate inhibitory effects of the extract obtained by the optimal technology on osteoclast differentiation. METHODS The index components of MT- 2 process optimization were selected by using network pharmacology. Based on single factor tests ，the extraction technology of MT- 2 was optimized by Box-Behnken design-response surface methodology according to the comprehensive score of contents of above index components ，and then validated. RAW 264.7 cells were induced by receptor activator of nuclear factor-κB ligand（100 ng/mL） to prepare osteoclast differentiation model. Inhibitory effects of MT- 2 extract（18.6，37.2，74.4 ng/mL）obtained by the optimal technology on osteoclast differentiation were investigated. RESULTS The index components screened by network pharmacology included chlorogenic acid ，terpineol diglucoside ，isochlorogenic acid A ，1，5-dicaffeoylquinic acid ，hydroxysafflower yellow A ， ginsenoside Rg 1 and ginsenoside Rb 1. The optimal extraction technology of MT- 2 was ethanol volume fraction of 60％ ，the solid-liquid ratio of 1 ∶ 14（g/mL），extraction time of 94 min and extraction times of twice. The average comprehensive score obtained by the three validation experiments was 95.50，and the relative error with the predicted value （95.75）was －0.26％. Compared with osteoclastic differentiation model cells ，the cells treated with MT- 2 extract prepared by the optimal technology were mostly mononuclear round cells ，and the number of osteoclasts decreased significantly （P＜0.05），its inhibitory effects tended to strengthen with the increase of drug concentration. CONCLUSIONS The optimal extraction technology of MT- 2 is stable and feasible. Obtained extract can inhibit osteoclast differentiation.
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OBJECTIVE To optimize the extraction technology of volatile components from Wuyao decoction. METHODS On the basis of single factor investigation ，the extraction technology of volatile components from Wuyao decoction was optimized and validated by Box-Behnken design-response surface technology using the contents of bomyl acetate ，cyperotundone，α-cyperone， ligustilide and dehydrocostuslactone ， extraction rate of volatile oil as indexes ， with extraction time ， soaking time and liquid-material ratio as factors. On this basis ，the extraction state of the decoction was quantified. RESULTS The optimal extraction technology was as followed ：the ratio of liquid -material was 13∶1（mL/g），soaking time was 0.5 h，and the extraction time was 6 h in the boiling state. The comprehensive scores of the three validation experiments were 0.948 7，0.948 4 and 0.948 6 respectively （RSD＝0.02％，n＝3），and the deviation from the predicted value （0.947 9）was no more than 1％. The boiling state of the decoction in 180 ℃ oil bath was taken as the sudden boiling state. CONCLUSIONS The optimized extraction technology is stable and feasible.
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OBJECTIVE To optimize the e xtraction technology of Guizhi shaoyao zhimu decoction （GSZD）. METHODS The contents of 9 components in GSZD were determined by HPLC ，such as ephedrine hydrochloride ，pseudoephedrine hydrochloride ， mangiferin，paeoniflorin，liquiritin，5-O-methylvisammioside，glycyrrhizic acid ，cinnamic acid ，6-gingerol. On the basis of single factor experiment ，taking material-liquid ratio ，extraction times and extraction time as inspection factors ，taking the contents of above 9 components and the yield of dry extract as evaluation indicators ，the analytic hierarchy process and entropy weight method were used to determine the composite weight of each index and calculate the comprehensive score ；the extraction technology parameters of GSZD were optimized by Box-Behnken response surface method ，and the validation tests were conducted. RESULTS The composite weight of the contents of ephedrine hydrochloride ，pseudoephedrine hydrochloride ，mangiferin，paeoniflorin， glycyrrhizin，5-O-methylvisa- midol ，glycyrrhizinate，cinnamic acid ，6-gingerol and the yield of dry extract were respectively 0.12，0.10，0.05，0.12，0.14，0.06，0.13，0.15，0.10，0.03. The optimal extraction technology of GSZD is that the ratio of material to liquid is 1 ∶ 14（g/mL），extraction is 2 times，and the extraction time is 3.0 h；average comprehensive score of the 3 verification tests was 95.879，and RSD was 0.50％（n＝3），the deviation from the predicted comprehensive score （94.328）was 1.64％. CONCLUSIONS In this study ，the optimal extraction technology of GSZD is determined.
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OBJECTIVE：To opt imize the extraction technology of phenolic acid from Amomum tsaoko . METHODS ：The extraction technology of phenolic acid from A. tsaoko was optimized by using Box-Behnken design-response surface methodology with ethanol volume fraction ，liquid-solid ratio and extraction time as factors ，using the total contents of protocatechuic acid and vanillic acid as response value. The optimizd extraction technology was vlidated. RESULTS ：The optimal extraction technology was as follows ：ethanol volume fraction 65％，liquid-solid ratio 4∶1（mL/g），extraction time 2.5 h. After 3 times of validation tests ， average total content of protocatechuic acid and vanillic acid were 12.32 mg/g（RSD＝0.26 ％，n＝3），average relative error of which with predicted value （12.63 mg/g）was 2.45％. CONCLUSIONS ：The optimal technology is stable and feasible .
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OBJECTIVE：To optimize the extraction technology of Zhideke granules. ME THODS：The extraction technology （water extraction ，alcohol extraction ，water extraction and ethanol precipitation ）of Zhideke granules was initially screened by ammonia-induced cough experiment and xylene-induced ear swelling experiment in mice. Based on its preparation route ，the immersion time of medicinal materials containing volatile oil was investigated with water absorption as index firstly. The single factor test was adopted to investigate the amount of water added and the extraction time taking the volatile oil yield as index to optimize the extraction technology of medicinal materials containing volatile oil. Taking the contents of irisflorentin and total flavonoids as indicators ，on the basis of single factor investigation ，orthogonal test was adopted to examine the influence of three factors including the amount of water added ，extraction time and extraction frequency ，so as to optimize the water extraction technology of Zhideke granules and the validation tests were conducted. RESULTS ：The results of pharmacodynamics experiment showed that the cough latency of mice in water extract low-dose and high-dose groups （6.34，12.68 g/kg，by crude drug ）and water-extraction alcohol-precipitation extract high-dose group （12.68 g/kg，by crude drug ）were significantly longer than those inmodel group ，and the number of cough within 2 minutes was significantly reduced （P＜0.05 or P＜0.01）. Compared with model group ， the ear swelling of mice in water extract low-dose and high-dose groups （6.34，12.68 g/kg，by crude drug），ethanol extract high-dose group （12.68 g/kg，by crude drug） and water-extraction alcohol-precipitation extract hig dose group （12.68 g/kg，by crude drug ） were decreased significantly （P＜0.05 or P＜0.01）. The swelling inhibition rates were 42.26％，55.08％，33.49％，51.56％，39.57％ and 44.36％ in low-dose and high-dose groups of water extract ，alcohol extract ， water-extraction and alcohol-precipitation extract respectively ，indicating that the water extract had better antitussive and anti-inflammatory effects. The optimal extraction technology of volatile oil was adding 5-fold water ，soaking for 30 minutes，and extracting for 3 hours. The optimal water extraction technology was adding 12-fold water ，extracting for 3 times after soaked for 50 min，lasting for 1 h each time. Results of 3 times of validation tests showed that average content of irisflorentin in the extract obtained by optimal technology was 76.47 μg/g（RSD＝ 2.15％，n＝3）and the average content of total flavonoids was 92.45 mg/g（RSD＝0.48％，n＝3）. CONCLUSIONS ：The optimal extraction technology of Zhideke granules is stable and feasible.
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To optimize the ethanol extraction technology parameters of Fengyin Decoction by orthogonal experiment combined with beetle antennae search(BAS)-genetic algorithm(GA)-back propagation neural network(BPNN). Based on single factor investigation, the extraction temperature, ethanol volume, extraction time, and ethanol concentration were used as orthogonal experiment factors, and entropy weight method was used to calculate the comprehensive scores of aloe-emodin, glycyrrhizic acid ammonium salt, rhein, emodin, chrysophanol, physcion, cinnamaldehyde, 6-gingerol, extraction ratio and fingerprint similarity. BAS-BPNN model was established, and then, GA was used to predict the optimal extraction process. The results showed that BAS-BPNN was optimized to obtain the optimal ethanol extraction process of Fengyin Decoction as follows: extraction temperature of 87 ℃, adding 9 times of 75 % ethanol, and extracting for 47 minutes, with a comprehensive score of 1.052 9. Meanwhile, the optimal process parameters obtained by orthogonal design were as follows: the extraction temperature of 80 ℃, adding 10 times of 75% ethanol, extracting for 30 minutes, with a comprehensive score of 1.003 7. The comprehensive score of the process obtained from the BAS-BPNN model was slightly better than that from the orthogonal test, indicating that the optimized process from BAS-BPNN model was more ideal, so it was finally determined as the best extraction process for Fengyin Decoction. The process of Fengyin Decoction obtained from BAS-GA-BPNN has high extraction efficiency and good stability, which provides reference for the subsequent development and quality control.
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Drugs, Chinese Herbal , Entropy , Ethanol , Neural Networks, Computer , Quality ControlABSTRACT
Objective: The extraction process optimization method of chemometrics combined with information entropy weight was established and applied to the water extraction process optimization of Banxia Baizhu Tianma Decoction (BBTD), in order to fully ensure the effectiveness and quality consistency of classical prescription. Methods: Taking BBTD as the model drug, the fingerprint was established by HPLC method. The common peak area was analyzed by principal component analysis (PCA), the total factor score of PCA, the similarity of fingerprint and the yield of dry paste were used as evaluation indexes. L9(34) orthogonal design was used to investigate the effects of adding water, soaking time and boiling time on the extraction effect, and the Objective: weight of each index was determined by information entropy weighting method. The technological parameters of water extraction were optimized. Results: Twenty-six common peaks and seven compounds were identified by the similarity evaluation system of chromatographic fingerprints of traditional Chinese medicine. According to the results of comprehensive score, the optimum extraction process of the preparation was determined to be 12 times the amount of water, decoction twice, 1 hour each time. The average comprehensive score of the three batches of verification was 0.418 0 with RSD of 3.32%. Conclusion: The optimized process has high extraction rate, good stability and repeatability, and is suitable for the industrial production of BBTD.
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Objective: To optimize the extraction conditions of baicalin from Scutellariae Radix by electromagnetic pyrolysis. Method: Based on single factor experiments, taking extraction time, material size and liquid-material ratio as factors, yield of baicalin as index, extraction parameters were optimized by response surface methodology, and compared with ultrasonic method, ethanol refluxing method and decoction method. Result: The optimal extraction conditions were as follows:extraction time of 2.41 min, material size of 100 mesh, liquid-material ratio of 33 mL·g-1. Under these conditions, the yield of baicalin was 12.21%. The yields of baicalin by ethanol refluxing method, decoction method and ultrasonic method were 12.91%, 12.62% and 11.61%, respectively. The yield of baicalin by electromagnetic cracking extraction was close to several other conventional extraction methods, and the extraction time was significantly shortened. Conclusion: As a novel extraction technology of traditional Chinese medicine, electromagnetic cracking extraction has the advantages of high efficiency, energy and time saving, green environmental protection, etc. And it can provide a new method for the industrial extraction of baicalin.
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Objective: To optimize the extraction technology of Guizhi Zhumian Capsules (GZC). Methods: Based on the analysis of the single factor experiment Results:, the orthogonal test method was adopted to study the three factors including the amount of water added, extraction time, and extraction frequency by taking the dry extract yield of the medicinal materials and transfer rate of geniposide as indicators to optimize the water extraction process of GZC. To optimize the alcohol precipitation process of GSC, these factors including the concentration of the medicinal materials, alcohol content, and time of alcohol precipitation were investigated. The extracts before and after alcohol precipitation were compared by improving sleep pharmacodynamics. Results: The best water extraction technology of GZC was decocted three times with 10 times of water, 0.5 h each time. The optimal alcohol precipitation process was to concentrate the filtrate of water extraction to 1 mL, which was equivalent to 1 g of the original medicinal materials, with 80% alcohol content and 12 h alcohol precipitation time. Compared with the negative control group, the water extract group of GZC could prolong the sleep time of pentobarbital sodium mice and increase the sleep rate of mice under the lower dose of pentobarbital sodium valve (P 0.05). Conclusion: The pharmaceutical efficacy of the extract before and after alcohol precipitation was different. In order not to affect the drug effect, the water extraction process was finally selected as the best extraction process.
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Objective: To optimize the extraction technology of Shenqi Qiangxin Tablets (SQT). Methods: With the improvement of heart lesion of pharmacological model of isoproterenol induced heart failure in rats as the index, pharmacological efficacy test was used to screen extracting conditions of the technology. The extraction technology was optimized by analytic hierarchy process combined with principal component analysis, single factor and orthogonal tests using each content of solid matter, ginsenosides Rg1, Re as indexes. And the verification test was carried out by using solid mass and icariin content as indexes. Results: Pharmacological efficacy test showed that technology 4 was superior. The optimal extraction condition of technology 4 was as follow: five medicinal materials including red ginseng and astragalus were reflux extracted three times with 50% ethanol, 11 fold for the first time, 10 fold for the second and three times, 2.5 h for each extraction; Epimedium and the other two medicinal materials were decocted three times with water, 19 fold for the first time, 16 fold for the second and third times, 1.5 h for each decction. The verification test showed that the average yield of ethanol extracted solids was 19.78%, and the average extraction rate of ginsenoside Rg1 and Re was 77.52%; The average value of water extracted solids was 16.58%, and the average extraction rate of epimedium was 90.98% (RSD < 2.0%, n = 3). Conclusion: The optimized extraction technology was stable and feasible.
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Objective: To optimize the extraction technology and inclusion process of volatile oil for Qingxiangrukang Granules (QG). Methods: Box-Behnken was used to optimize the extraction technology of volatile oil in QG. With yield of inclusion compound and inclusion rate of volatile oil-β-CD as evaluation indexes, and with volatile oil-pure water ratio, β-CD-volatile oil ratio, and inclusion time as investigate factors, the optimal inclusion technology for volatile oil of QG was ensured based on PCA-G1-entropy method and orthogonal design and by using colloid milling. Results: The optimum extraction technology of volatile oil were as follows: extracting time was 6 h, liquid-material ratio was 10, immersion was 0 h. The optimal inclusion technology for volatile oil were as follows: pure water-volatile oil ratio was 1:80, β-CD-volatile oil ratio was 6 g, and the inclusion time was 30 min. Under such condition, there was no significant difference between verification groups of three batches. Conclusion: This optimal extraction technology and inclusion process is stable and workable and can provide experimental basis for industrial production of QG.
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Objective: To optimize the water extraction technology parameters of Yiqi Huoxue Prescription (YHP). Methods: On the basis of single factor experiment, orthogonal experiment design was used to evaluate the transfer rate and extraction yield of salvianolic acid B and hydroxysafflower yellow A by using adding water, extraction time and soaking time as factors. The comprehensive score was obtained by G1-entropy weight method. The optimal water extraction technology was obtained by orthogonal test design, and another method-BP neural network modeling was used to optimize the network model and target optimization. The two analytical methods were compared in the verification experiment to find the optimal water extraction technology parameters of YHP. Results: Based on the comparison of the two analytical methods, it was found that the comprehensive score of the optimal water extraction technology obtained by orthogonal test analysis was slightly higher than that obtained by BP neural network modeling. Therefore, it was finally determined that the optimal water extraction technology parameters of YHP were as follow: water extraction for three times, soaking for 0.5 h, adding water of 20 times, and extracting time for 3.5 h. Conclusion: The optimal water extraction technology of YHP is stable and feasible, which provides new ideas and references for the development and modernization of new drugs of compound Chinese medicine.
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Objective: To optimize the extraction of total flavonoids from leaves of Psidium guajava. Methods: L9(34) orthogonal test was used to screen the optimal extraction technology of total flavonoids from leaves of P. guajava with ethanol concentration, solid-liquid ratio, extraction time, and extraction times as factors. The weight coefficient of the four evaluation indicators, including the yield of total flavonoids, hyperoside, quercetin-3-O-β-D-pyranarabinoside, and quercetin-3-O-α-L-furanarabinoside was calculated by information entropy weighting so as to calculate comprehensive score. We obtained the optimal technology by orthogonal analysis based on the comprehensive score. Results: The optimal extraction technology was that using 8-fold 70% ethanol water to extract 2 h for three times. The mean comprehensive score of the three batches was 0.142 1 and the RSD was 2.37%. Conclusion: The optimal extraction technology of total flavonoids from leaves of P. guajava was stable and feasible with high yield.
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OBJECTIVE： To establish the method for content determination of total flavonoids from Combretum alfrdii， and to optimize the extraction technology of total flavonoids from C. alfrdii. METHODS： Using aluminium trichloride as， chromogenic agent， UV spectrum was adopted to determine the content of total flavonoids from C. alfrdii. Based on single factor test， ethanol volume fraction， material-liquid ratio， extraction time， extraction temperature and times were selected as investigation factors， and the content of total flavonoids was selected as response value， Plackett-Burman design was used to screen the factors that had significant influence on the content of total flavonoidsfrom C. alfrdii. Then steepest climbing test was adopted to confirm the optimum valuing range； the extraction technology of total flavonoids was optimized by Box-Behnken response methodology. RESULTS： The linear range of total flavonoids were 0.012-0.036 mg/mL （r＝0.999 9）； RSDs of precision， stability and repeatability tests were less than 3％； the recovery ranged from 92.98％ to 99.86％ （RSD＝2.71％， n＝6）. The optimal extraction technology included that 60％ ethanol， material-liquid ratio of 1 ∶ 34 （g/mL）， extracting for 3 times， lasting for 60 min， extraction temperature of 80 ℃. Under this technology， average content of total flavonoids from C. alfrdii was 2.71％ （RSD＝1.69％， n＝6）， and the relative error was 2.65％ compared with predicted value of the model （2.64％）. CONCLUSIONS： Established method is stable and reproducible， and can be used for content determination of total flavonoids from C. alfrdii. The optimized extraction method is stable and feasible.
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OBJECTIVE： To optimize the water extraction technology of Bupi yangshen granules， and to provide basis for the follow-up research and development of it. METHODS： The contents of astragaloside Ⅳ and salvianolic acid B in water extract of Bupi yangshen granules， were determined by HPLC-ELSD and HPLC-DAD. Using the comprehensive score of contents of astragaloside Ⅳ and salvianolic acid B and extract yield as index， weight coefficient of indicators were determined by AHP， CRITIC and AHP-CRITIC mixed weighting method. L9（34） orthogonal test was used to optimize decoction time， water volume and decoction times in water extraction technology of Bupi yangshen granules. Validation test was also performed. RESULTS： The weight coefficient determined by AHP-CRITIC mixed weighting method was the most reasonable. The optimal extraction technology was decocting twice， adding 12-fold water， 1 h each time. The results of 3 times of validation test showed that the average contents of astragaloside Ⅳ and salvianolic acid B were 8.79， 609.50 mg （total amount of 121 g medicinal herbs extracted from whole prescription）， respectively. The average extract yield was 31.24％. Average comprehensive score was 96.59（RSD＝1.01％，n＝3）. CONCLUSIONS： The optimized water extraction technology is reproducible， stable and feasible. It can provide a scientific basis for the follow-up development and industrial production of Bupi yangshen granules.
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OBJECTIVE： To establish the method for simultaneous determination of saikosaponin a and saikosaponin d in Bupleurum chinense water extract， and to optimize its water extraction technology for electromagnetic cracking. METHODS： HPLC method was used. The determination was performed on SB-C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 40 ℃. The detection wavelength was set at 210 nm， and the sample size was 10 μL. Based on single factor experiment， using extraction time， particle size， solide-liquid ratio as factors， total extraction rate of saikosaponin a to saikosaponin d as indexes， the extraction technology was optimized by using Box-Behnken response surface methdology， and compared with the results of ultrasound method and decoction method. RESULTS： The linear range of saikosaponin a and saikosaponin d were 50.70-202.80 μg/mL （r＝0.999 9） and 50.50-202.00 μg/mL （r＝0.999 9）， respectively. The quantitation limits were 0.16 and 0.13 μg/mL， respectively. The detection limits were 0.05 and 0.04 μg/mL，respectively. RSDs of precision， stability and reproducibility tests were all lower than 2％. The average recoveries were 98.23-102.47％ （RSD＝1.80％， n＝6） and 98.84％-102.06％ （RSD＝1.60％， n＝6）. The optimal extraction technology was as follows： the extraction time of 2.50 min； the particle size of 80 mesh， solid-liquid ratio of 1 ∶ 28 （g/mL）. Results of 3 times of validation tests showed that the optimal technology included the average total extraction rates of saikosaponin a and saikosaponin d were 8.42 mg/g， which was higher than that of ultrasonic method （8.34 mg/g） and decoction method （8.06 mg/g）， and the extration time was shorter. CONCLUSIONS： Established method is simple and accurate， and can be used for simultaneous determination of saikosaponin a and saikosaponin d in B. chinense water extract. The optimized water extraction technology for electromagnetic cracking is stable and feasible.
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OBJECTIVE： To establish a determination method for content of polysaccharide in Acanthopanax senticosus， and to conduct cluster analysis and ultrasonic extraction technology optimization. METHODS： The content of polysaccharide in A. senticosus was determined by phenol-sulfuric acid method. Cluster analysis was conducted by using SPSS 23.0 software of the polysaccharide in A. senticosus from 17 habitats. The ultrasonic extraction technology was optimized with L9（34）orthogonal test using extraction temperature， the ratio of material to liquid， extraction time as factors， the content of polysaccharides as index， and then validated. RESULTS： The linear range of glucose ranged from 0.007 75 to 0.151 mg/mL （r＝0.999 1）； the limits of quantification and detection were 2.854， 0.856 μg/mL； RSDs of precision， stability， and repeatability tests were less than 2％； recovery rates of the sample were 98.41％-101.58％（RSD＝1.23％，n＝6）. Cluster analysis results showed that 17 batches of samples could be clustered into three classes； S1， S6， S10， S12 and S13 were clustered into one class； S3-S5 and S7 were clustered into one class； and the rest samples were clustered into one class. The optimal ultrasonic extraction technology was extraction temperature 55 ℃， ratio of material to liquid 1 ∶ 10 （g/mL）， extraction time 35 min. Results of validation tests showed the content of the polysaccharide under the best process condition was 4.36％（RSD＝0.920％，n＝3）. CONCLUSIONS： Established method is simple，reproducible and suitable for determination of polysaccharide in A. senticosus； the optimized ultrasonic extraction technology is stable and feasible.
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OBJECTIVE： To optimize the extraction technology of verbascoside from Cistanche tubulosa， and to provide reference for further development and comprehensive utilization of C. tubulosa. METHODS： The content of verbascoside in C. tubulosa was determined by HPLC. The determination was performed on Inertsil-ODS-3V column with mobile phase consisted of methanol-0.2％ formic acid aqueous solution （40 ∶ 60， V/V） at the flow rate of 1 mL/min. The column temperature was 30 ℃， the detection wavelength was 330 nm， and the sample size was 10 μL. Using extraction rate of verbascoside as index， soaking time， ethanol concentration， liquid-solid ratio， extraction time and extraction times were investigated by single factor tests. According to the results of above tests， ethanol concentration， liquid-solid ratio and extraction time were optimized by Box-Behnken response surface methodology. The verification tests were carried out on the optimized extraction technology. RESULTS： The linear range of verbascoside was 18.65-932.4 μg/mL. The optimal extraction technology included that ethanol concentration 63％， liquid-solid ratio 8 ∶ 1 （mL/g）， soaking for 2 h， extraction time 1.5 h， extracting for 2 times. The extraction rates of verbascoside in the three parallel verification tests were 78.21％， 76.95％， 79.34％， respectively. The relative errors of those to predicted value 76.76％ were 1.89％， 0.25％， 3.36％. CONCLUSIONS： The optimized extraction technology of verbascoside from C. tubulosa is stable and feasible， and is suitable for the extraction of verbascoside.
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OBJECTIVE： To establish a method for the content determination of total flavonoids from Typhonium divaricatum， and to optimize its extraction technology. METHODS： The content of total flavonoids in T. divaricatum was determined by UV spectrophotometry. Using the extraction amount of total flavonoids from T. divaricatum as index， the volume fraction of ethanol， the ratio of material to liquid， the extraction time and times as factors， the extraction technology of total flavonoids from T. divaricatum was optimized by L9（34） orthogonal design， based on the single factor test. RESULTS： The linear range of rutin were 8-48 μg/mL （r＝0.999 7）； the quantification limit was 0.54 μg/mL， and the detection limit was 0.18 μg/mL； RSDs of precision， stability and repeatability tests were all lower than 2％. The recoveries were 99.61％-102.38％（RSD＝1.15％， n＝6）. The optimal extraction technology was as follows： ethanol concentration of 70％， solid-liquid ratio of 1 ∶ 20 （g/mL）， extraction time of 45 min， extracting for 2 times. Under this condition， the average content of total flavonoids from T. divaricatum was 2.74 mg/g. CONCLUSIONS： Established method is simple and accurate； the extraction process is stable and feasible.