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ObjectiveTo optimize the extraction and purification process of Gardeniae Fructus for industrial production， and to obtain the total iridoid and total crocin extracts. MethodOrthogonal test was used to optimize the water extraction process by taking contents of geniposide， genipin gentiobioside， gardenoside， crocin-1 and crocin-2 as indicators and the decocting time， decocting times and water amount as factors. The purification process was optimized by single factor test， and four different types of macroporous adsorption resins were screened. The process conditions such as resin type， maximum loading amount， water washing amount， ethanol concentration， ethanol dosage， and flow rate of sample loading were mainly investigated. In addition， the drying methods （vacuum drying and spray drying） of the extract were investigated， and a pilot scale-up verification test was carried out. ResultThe optimal water extraction process of Gardeniae Fructus was to add 15， 10 times the amount of water for decocting twice， 1 h each time. The optimal purification process was as follows：the water extract through SP825L macroporous resin column， the amount of crude drug-the amount of resin （1∶1.5）， the sample loading flow rate of 3 BV h-1， adding 2 BV of water to remove impurities， adding 4 BV of 30% ethanol to obtain the iridoid part， then adding 3 BV of 70% ethanol to obtain the crocin part， collecting the ethanol lotion， and drying at 70 ℃. Under these conditions， the extraction amount of total iridoids was 590.75 mg·g-1 with the transfer rate of 70.48%， and the yield of dry extract was 8.89%. The extraction amount of total crocins was 83.37 mg·g-1 with the transfer rate of 22.20%， and the dry extract yield was 2.60%. ConclusionThe optimized extraction and purification process is stable and feasible with high extraction rate of active components， which is suitable for the industrial extraction and purification of active parts of Gardeniae Fructus.
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Objective To screen the macroporous adsorption resin suitable for the separation and purification of total polyphenols from purple tea and establish the purification process parameters to prepare high-purity total polyphenols from purple tea. Methods The static adsorption-elution test was used to screen macroporous adsorption resin for the purification of total polyphenols from purple tea. Based on the single factor test, the comprehensive score of adsorption rate was used as the index to investigate the effects of different factors on the purification process and identify the optimal parameters for the purification process. Those factors included sample concentration, the pH value of the sample solution, the ratio of column diameter to height, sample size, ethanol percentage in the eluent, eluent volume and elution flow rate. Results The best process parameters for purification of total polyphenols from purple tea by AB-8 macroporous adsorption resin were as following. The sample concentration was 375 μg/ml with flow rate 2 ml/min. The sample volume was 3 BV. The sample solution pH was 2. The ratio of colume diameter to height was 1∶6. The impurities were removed first by water 3 BV. 50% ethanol 4 BV was used for elution with flow rate 2 ml/min. Conclusion AB-8 macroporous resin was selected for the purification of polyphenols from purple tea under the optimized technological conditions. The mass fraction of total polyphenols increased from 40.2% to an average of 69.8%. The solid content decreased from 56.0 mg to 29.9 mg. The established purification process has good stability and feasibility. It can be used as a purification process for total polyphenols from purple tea.
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In recent years, macroporous adsorption resin has been widely used in the purification and separation of natural medicine and effective components of Chinese materia medica, the purification of compound Chinese medicine preparations, the removal of harmful impurities due to the advantages of stable physical and chemical properties, high selectivity, strong adsorption capacity and easy elution, recyclable use and regeneration treatment, economic and environmental protection, convenient preparation molding and so on. By summarizing the preparation, properties, classification and working principle of macroporous adsorption resin, the influencing factors and purification process of Chinese materia medica components were reviewed. The purpose of this paper is to explore the important factors affecting the purification of Chinese materia medica components by macroporous adsorption resin, and provide reference for improving the purification effect of Chinese materia medica components.
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Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex,the process parameters included concentration of sample solution,speed of sampling,diameter-height ratio of resin column,water washing amount,concentration and dosage of eluent,flow rate of elution,etc.In order to determine the optimum process,HPLC was employed to determine the contents of four alkaloids(magnoflorine,jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm.After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following:10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1:8,sampling solution concentration of 11 g·L-1,the loading flow rate of 1 mL·min-1,sampling solution volume of 50 mL,washed with 4 BV of water(1 BV=15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was>80%,and its purity was>65%.The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was>0.99. Conclusion: This optimized purification process is stable and feasible, and the established quality standard is controllable.
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Objective To screen the macroporous adsorption resin suitable for the separation and purification of total flavonoids from Litchi Semen, and the purification process parameters were established to prepare the total flavonoids of Litchi Semen in accordance with the requirements of effective parts of Chinese materia medica, which laid the foundation for the development of the total flavonoids of Litchi Semen into five new Chinese medicines. Methods The macroporous adsorption resin for purifying the total flavonoids of Litchi Semen by static adsorption-elution test was used. Based on the single factor test, the comprehensive score of adsorption rate was used as the index to investigate the volume fraction of ethanol, the mass concentration of the sample, and the sample solution pH, diameter to height ratio, upper column volume, upper column flow rate, eluent concentration, eluent volume and elution flow rate on the purification process, and determine the optimal purification process parameters. Results The best process condition for separating the total flavonoids of Litchi Semen by AB-8 macroporous adsorption resin were as follows: the mass ratio of resin to medicinal material was 3:1, the concentration of the upper column sample solution was 4—6 mg/mL, sample flow rate was 1 mL/min, and the upper column volume was 2 BV, diameter to height ratio was 1∶12, pH of the sample solution was 2, first impurity removal by 20% ethanol 3 BV, and using 60% ethanol 3 BV for elution, elution flow rate was 4 mL/min. Conclusion AB-8 macroporous resin can be used to purify the total flavonoids of Litchi Semen under the established technological conditions. The mass fraction of total flavonoids in Litchi Semen increased from 29.22% to an average of 67.37%, and the solid content decreased from 1.25 g to 0.40 g. It indicates that the established purification process is stable and feasible, and can be used as a purification process condition for total flavonoids of Litchi Semen.
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Objective: To investigate the adsorption performance and purification effect of macroporous adsorption resin on total polyphenols in Acanthopanan trifoliates leaves, and to determine the technological conditions for the purification of total polyphenols from A. trifoliates leaves. Methods: The Folin-Ciocalteu colorimetric method was used to quantify the adsorption and desorption effects of five macroporous adsorption resins on the total polyphenols in the A. trifoliates leaves. The resin suitable for separation and purification of total polyphenols in A. trifoliates leaves was screened, and the adsorption and desorption conditions were investigated and optimized. The final optimized parameters were determined by single factor experiments. Results: The best purification parameters of total polyphenols were determined as follows: the concentration of sample solution was 1.0 mg/mL (crude drug) with pH 3.0, sample flow rate was at 2 mL/min, and the sample loading was controlled to 30 mL, the elution was 50% ethanol at a flow rate of 2.0 mL/min with pH 6.0, and the elution amount was 40 mL. The polyphenol sample of the A. trifoliates leaves was purified by HPD100 resin, and the purity increased from 11.7% to 49.7%, and the purification effect was 4.25 times than before. After 30 times enlargement experiment, the purity of the A. trifoliates leaves polyphenol sample increased from 12.5% before purification to 54.5%, and the purification effect was 4.4 times than before. The amount of macroporous resin did not affect the purification efficiency, which provided reference for HPD100 macroporous resin for industrial production of total polyphenols purification of A. trifoliates leaves. Conclusion: HPD100 is the best resin for purifying total polyphenols from A. trifoliates leaves, and the process technology result in this experiment can be applied to industrial production.
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Objective: To optimize the purification technology of saponins in steamed Panax notoginseng with macroporous resin. Methods: The main factors affecting the purification process were screened by failure mode and effects analysis (FMEA). The purification method with macroporous resin was optimized by central combination design-response surface method (CCD-RSM) based on the recovery and purity of saponins. In this experiment, the concentration of sample solution, loading volume, washing volume, ethanol concentration, and ethanol elution volume were used to investigate the purification of saponins in steamed P. notoginseng. Results: The optimized purification process with macroporous resin was as follows: maximum recovery (82.81%) and purity (77.24%) of saponins were obtained with the concentration of saponin solution of 11.22 mg/mL, loading volume of 4.97 BV, washing volume of 2 BV, ethanol concentration of 70%, and ethanol elution volume of 3.31 BV. Conclusion: The optimized purification process based on FMEA and CCD-RSM is convenient and stable, with high recovery and purity of saponins, which has a certain practical value.
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Objective To explore the effects of chitosan flocculation clarification process and alcohol precipitation process on the principal chemical constituents of water extract of Codonopsis Radix. Methods The influence of two purification processes on water extract of Codonopsis Radix was investigated through lobetyolin contents, extract yield, and relative apparent content of each component in HPLC fingerprint as evaluation indexes. Results Chitosan flocculation clarification process showed a significantly higher extract yield of water extract compared with alcohol precipitation process, and it has a markedly better retention effect for strong polarity constituents; alcohol precipitation process exhibit a little better retention effect for lobetyolin and a better retention effect for weak polarity constituents. Conclusion The above two processes have some regularity in the influence on the main chemical constituents in the water extract of Codonopsis Radix, which can provide some guidance for the reasonable choice of the purification process for water extracts of Codonopsis Radix, and other TCM water extracts.
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Objective To investigate the optimal process conditions for the separation and purification of extract from Hyssopus cuspidatus Boriss. by polyamide resins. Methods The total flavonoids and rosmarinic acid were used as the indexes. The maximum amount of sample solution, elution volume, concentration of sample solution, adsorption time of resin, loading time of sample solution and the amount of eluting solvent, pH and elution rate in the resin purification process were screened by single factor method. Results The optimal purification parameters were as follows: 10 mg/mL of extract, 12 mL of sample amount, 2 BV of water to remove impurities, 40% ethanol to elute 9 BV; the concentration of rosmarinic acid in sample solution was 86.3 μg/mL, and the total flavonoid concentration was 117.8 μg/mL; the resin adsorption time was 14 h; the pH of sample solution was 6.5; the elution rate was 3.0 BV/h. Conclusion This method is simple and feasible, fit for separating and purifying of extract from Hyssopus cuspidatus Boriss.
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Objective To multi-objectively optimize the purification process parameters of Yinju Jiedu Oral Liquid (YJOL), mixed weight of AHP-CRITIC, Plackett-Burman design (PBD), and Box-Behnken design (BBD) were adopted on the basis of HPLC fingerprints. Methods HPLC was used to establish HPLC fingerprint of YJOL. Recovery rate of fingerprints of six components (chlorogenic acid, linarin, harpagoside, R,S-epigoitrin, psoralen, and isobalin) and HPLC fingerprint similarity was taken as the index to optimize the type of macroporous resin among 10 types. Weights of the recovery rates of six components and similarity were determined by mixed weight of AHP-CRITIC, in order to obtain the comprehensive index as the evaluation criterion. The significantly influencing factors were firstly evaluated by PBD, and purification conditions were optimized by BBD. Results HPD-400 type resin showed a high selectivity for six components. The optimized purification technology was as follows: the ratio of dia-height was 1:7, pH value was 3.5, sample concentration was 0.18 g/mL, ratio of sample to resin was 0.96 g/g, and 77% ethanol's dosage was 8 BV. Under the conditions, the recovery rates of six components were 78%-98%, and the similarity of fingerprint was higher than 0.99. These components could get balanced recovery. Moreover, the theoretical and actual comprehensive indexes were 94.28% and 93.69%, respectively, with a relative error of 0.59%. Conclusion Based on HPLC fingerprints, mixed weight of AHP-CRITIC, combined with PBD and BBD-RSM used to optimize the purification process for the YJOL in this study is scientific and feasible, this way can improve the purity of the active components and keep the uniformity of main components in the YJOL as well.
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OBJECTIVE: To study the purification process of anthocyanins from purple Solanum tuberosum by AB-8 macroporous resin combined with alcohol precipitation method. METHODS: Using River John Blue genotype purple Solanum tuberosum as raw material, purple Solanum tuberosum anthocyanin crude extract was obtained by leaching method. Then, the purification process of the crude extract was studied by using static adsorption (stirring, loading concentration, loading pH, eluent concentration, eluent pH) and dynamic adsorption (loading flow rate and elution flow rate), respectively, taking adsorption rate, resolution rate and color value as the evaluation indexes. On the basis of the single factor experiment, orthogonal optimization analysis was carried out to get the final optimized process condition. RESULTS: The optimum conditions for the purification of AB-8 macroporous resin were as follows stirring, loading concentration 0.500 mg·mL-1, loading pH 2, ethanol concentration 70% and elution pH 1. The column flow rate was 1 mL·min-1, and the elution flow rate was 1 mL·min-1. Under this condition, the color value of anthocyanin was able to reach 75.40, which was 8.43 times higher than that before purification. CONCLUSION: The purification method has high feasibility and can improve the purity and color value of anthocyanins, which laids a foundation for the market development and application of anthocyanins of purple Solanum tuberosum.
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OBJECTIVE:To optimize the purification process of the extract from Yiqi guben granules. METHODS:The purifi-cation effect of the process was investigated with transfer rate of polysaccharide,calycosin glucoside and dry paste as evaluation in-dexes,using ZTC1+1 type Ⅱ,and shell poly sugar and 95% ethanol as clarifying agents. The purification process of the extract from Yiqi guben granules was optimized by orthogonal test using the ratio of material to liquid,the amount of clarifying agent and standing time as factor. The validation test was conducted. RESULTS:Selecting ZTC1+1 type Ⅱ as a clarifying agent,the best trans-fer rate of effective component had been obtained;optimal purification process was as follows as the ratio of material to liquid 1:2, the ratio of ZTC1+1 type Ⅱ A liquid 5%,the ratio of B liquid 10%,standing time of 5 h. The results of verification test showed transfer rates of dry paste in 3 tests were 71.54%,70.98%,69.21%,respectively;those of polysaccharide were 82.55%, 81.78%,82.15%,respectively;those of calycosin glucoside were 91.92%,92.34%,91.58%,respectively (all RSD≤1.72%, n=3). CONCLUSIONS:The optimized purification process is effective,stable and practical,and can be used for the purification of the extract from Yiqi guben granules.
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Objective: To multi-objectively optimize the purification process parameters of licorice flavonoids using entropy-weight method, Plackett-Burman design (PBD), and Box-Behnken design (BBD). Methods: On the basis of HPLC fingerprints of licorice, the macroporous resin type was chosen using the recovery rates of six components (liquiritin apioside, liquiritigenin, isoliquiritin apioside, licuraside, isoliquiritin, and neoisoiiquiritin) as detection indexes. Weights of the recovery rates of six components were determined by entropy-weight method, in order to obtain the comprehensive index. The significantly influencing factors were firstly evaluated by PBD, then purification conditions were optimized by BBD. Results: ADS-7 type resin showed a high selectivity for six components. The optimized purification technology was as follows: pH value was 4.5, sample concentration was 0.20 g/mL, ratio of sample to resin was 1.0 g/g, flow rate was 0.6 mL/min, elution dosage was 6.7 BV, ethanol concentration was 83% of eluting agent, and elution rate was 1.0 mL/min. Under the conditions, the recovery rates of six components were 86%-97%, the theoretical and actual comprehensive indexes were 93.32% and 93.05%, respectively, with a relative error of 1.17%. Conclusion: Entropy-weight method combined with PBD and BBD-RSM used to optimize the purification process for the licorice flavonoids in this study is scientific and feasible, providing a new reference to realize the multi-objective optimization of the purification technology for active constituents in Chinese materia medica.
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OBJECTIVE: To optimize the extraction and purification process of eurycomanone (EN) from Eurycoma longifolia Jack. METHODS: Using single factor test and orthogonal test to screen the best conditions of ethanol concentration, extraction time, amount of solvent and extraction times with the index of transfer rate of EN. Resin model was screened by static adsorption and desorption test, purification process of EN was investigated by single factor test and orthogonal test. RESULTS: The optimal condition of the extracting was 20-fold water, 2 times with the first 2 and 1 h for the second time. Optimal purification process of HPD100 macroporous resin was 70% amount of saturated adsorption sample, 0.25 g·mL-1 of sample liquid concentration, 3BV·h-1 of sample flow rate, eluting using the 30% ethanol, 3 BV·h-1 of elution velocity. CONCLUSION: The optimized process is simple, stable and repeatable, which is suitable for industrial production.
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Objective: Through the grey correlation analysis technology and the effect of Caco-2 colon cancer cell proliferation to evaluate the optimization of purification process in total flavonoids of Schizonepetae Herba by macroporous adsorption resin. Methods: By static adsorption and desorption rate of total flavonoids as examining indexes to screen the ratios of macroporous adsorption resin, using single factor experiment to optimize its purification process conditions of total flavonoids; The Caco-2 cell inhibition rate was determined by MTT method; At the same time, using the grey correlation degree analysis technology, the correlation between the purity of total flavonoids and proliferation inhibition rate of the tumor cell was calculated. Results: HPD-400 macroporous adsorption resin has the good adsorption separation performance of the total flavonoids from Schizonepetae Herba, Schizonepetae Herba at the concentration of 0.1 g/mL, 0.8 g/mL (medicine/wet resin) on the amount of column, using 2 BV water to remove impurity, eluting by 9 BV 70% ethanol and all of the flow rate are 4 BV/h, collecting the eluent and dry it; With the same method to do the secondary purification, the purity of total flavonoids is 92.15%, the inhibitory rate of Caco-2 cell proliferation is 86.17%, the greycorrelation coefficient of flavonoids purity and inhibition is 0.9502. Conclusion: This optimization process and the conditions are feasible and stable, the inhibitory efficacy of the in vitro Caco-2 cells is clearly. It lays the foundation of the new anti-cancer drugs based on the purification of total flavonoids in Schizonepetae Herba.
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Objective To compare the purification process of two types of ceramic hydroxyapatite(CHT I and CHT II)to remove the aggregates from two monoclonal antibodies(mAb 1 and mAb 2).Methods All the chromatography runs were performed on AKTA AVANT 150 with Tricon 10/50 column.The dynamic binding capacity( DBC) of two types of CHT was studied firstly, and then purification research was carried out selecting the suitable DBC.The column was equilibrated with 5 mmol/L sodium dihydrogen phosphate pH 6.5, and then was eluted with gradient buffers which were 10 mmol/L sodium dihydrogen phosphate pH 6.5 and 2 mol/L sodium chloride pH 6.5.Aggregate content in loading and elution pool was evaluated by size exclusion chromatography.Scale-up process was carried on 20 cm height chromatography column XK16/40.Results DBC of CHT I for mAb 1 was 40 mg/mL and mAb 2 was 45 mg/mL.After purity, monomer content of mAb 1 reached 98.6% and yield was 92.5% and monomer content of mAb 2 reached 98.8%and yield was 91.5%.DBC of CHT II for mAb1 was 16 mg/mL and mAb 2 was 20 mg/mL.After purity, monomer content of mAb 1 reached 99.8% and yield was 91.8% and monomer content of mAb 2 reached 99.9% and yield was 92.2%.Conclusion Two types of CHT both can remove aggregates effectively from monoclonal antibodies when aggregate content reaches more than 10%, and results conform to the regulations.CHT I has higher dynamic binding capacity than CHT II, and CHT II is superior to CHT I in removing aggregate efficiency.The purification process is simple and can be easily scaled up in pilot and manufacture.Therefore, it meets the requirement pilot and scale production.
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This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.
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OBJECTIVE: To optimize the extraction and purification process of total triterpenoid acid in Ligustri lucidi fructus using response surface method and uniform test. Finally, the content of total triterpenoid acid reached more than 50%. METHODS: The content determination of total triterpenoid acid in Ligustri lucidi fructus was established by HPLC. Use ethanol as extraction solvent and through the single factor investigation about the ethanol concentration, solvent amount, extraction time and extraction times, we determined the number of ethanol extraction. In addition to extraction times, the other three factors were further optimized by response surface method. Then purified by the alcohol extraction-water precipitation method to remove the water-soluble ingredients. Petroleum ether was used to remove the chlorophyll, grease and other small polarity ingredients; using ethanol to remove the alcohol soluble components whose polarity is bigger than fructus ligustri lucidi terpene acids. Design the uniform experiment respectively to determine the amount of petroleum ether, frequency and time, also ethanol concentration and dosage. RESULTS: The content of oleanolic acid and ursolic acid is larger than other compounds in total triterpenoid acid in Ligustri lucidi fructus, and they are isomers. When the results are in the equivalent of oleanolic acid, the optimum extraction process was determined as 80%, 10 times the amount of ethanol, extracting for 2 times, each time 0.5 h. The optimal purification process is 12 times the amount of petroleum ether, purified once, 2 h every time. Then 10 times amount of 60% ethanol to purify for once. CONCLUSION: Optimizing the extraction and purification process of total triterpenoid acid in Ligustri lucidi fructus by using response surface method and uniform experiment is simple, feasible and worth promoting.
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Objective: To optimize the purification process of celandine in Celandine Alkali from Tong'an Injection, and to improve the content of celandine in Celandine Alkali extract. Methods: In order to select the best way to separate celandine acid liquid, a study was carried out in three different ways of separation such as plate straight association-like centrifuge, high speed tubular centrifuge, and disc centrifuge; We investigated the dissolving effect caused by different pH values of hydrochloric acid to celandine dry extract; Three separation methods on celandine acidic lysate were investigated to choose the best, such as vacuum filtration, high-speed centrifuge tube, and flat direct-coupled centrifugal. Celandine Alkali transfer rate, and dry paste rate were as used evaluation indexes, so as to determine the feasible celandine refining purification process. Results: It was the best way to separate celandine acid liquid by using disccentrifuge; Hydrochloric acid could have the best dissolving effect on celandine dry extract when the pH values were 3-4, and the transfer rate of chelidonine was also the highest; High-speed tubular centrifuge was the best separation method for the celandine acidic dissolution liquid. Conclusion: The optimized process is simple, stable and viable, and it could be used for the preparation of qualified celandine intermediates.
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OBJECTIVE: To optimize the purification process of 3-O-[β-D-xylopyranosyl-(→3) -α-L-rhamnopyranosyl-(1→2)-α-L-arabinpyranosyl] -28-O-[α-L-rhamnopyranosyl-(1→4) β-D-glucopyranosyl -(1→6)-β-Z)-glucopyranosyl]-hederagenin (HE-III) from Nigella glandulifera Freyn et Sint by macroporous resin. METHODS: Resin model was screened by static adsorption and desorp-tion test, and the purification process of HE-III was investigated by single factor test and orthogonal test. RESULTS: The optimum purification process of AB-8 macroporous resin was as follows: the concentration of sample liquid was 0.25 g·mL-1, sample flow rate was 1.5 BV·h-1, the eluent was 70% ethanol, and the elution velocity was 3 BV·h-1. The content of HE-III in the intermediate and yield rate were 47.97% and 76.26%, respectively. CONCLUSION: The optimized process is simple and stable with good repeatability, which is suitable for industrial production.