CN105367580A - Preparation method of corydalis amabilis alkaloid monomers - Google Patents
Preparation method of corydalis amabilis alkaloid monomers Download PDFInfo
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Abstract
The invention discloses a preparation method of corydalis amabilis alkaloid monomers. The preparation method includes the following steps that corydalis amabilis extract is separated and purified through high-speed centrifugation partition chromatography, and then the corydalis amabilis alkaloid monomers are obtained. According to the preparation method, all the corydalis amabilis alkaloid monomers can be prepared on a large scale and can reach the gram level to the maximum degree, and the prepared monomers are high in purity which can reach 98% or above. The preparation method is low in cost, high in separation efficiency and recovery rate, high in product purity and large in preparation quantity.
Description
Technical field
The present invention relates to a kind of preparation method of Rhizome of Decumbent Corydalis alkaloid monomer.
Background technology
The stem tuber of papaveracease of doing nothing summer Corydalis Subgen. Capnites plant Hotseason grow Corydalisdecumbens (Thunb.) Pers..Have another name called a Jindan, open country prolonged recklessly, angle, Yanhusuo of throwing oneself on the ground, overboard pearl in hole.Just this medicine (Jindan) is described: " angle in a hole, have another name called open country and prolong recklessly, people from south of the River exhales sudden tree peony " in Qing Dynasty's Zhao Xuemin supplementary Amplifications of the Compendium of Materia Medica.
Rhizome of Decumbent Corydalis plant-growth in hills or hillside, low mountain, meadow, happiness be born in warmer climate humidity, on the sunny side, good, the soil layer of draining deep sandyly.Main product in Jiangxi, Hunan, Fujian, Zhejiang, the province such as Anhui, be good with Jiangxi yield and quality.Rhizome of Decumbent Corydalis is a kind of conventional Chinese medicine among the people, has the effect of analgesia, anti-inflammatory, is used for the treatment of stomachache, pain caused by cancer, sciatica, rheumatic arthritis, poliomyelitis sequela.Modern age, pharmacological research found that Rhizome of Decumbent Corydalis demonstrated good curative effect in cardiovascular and cerebrovascular diseases, as treatment senile dementia, anti-arrhythmia, anticoagulant etc.
The activeconstituents of Rhizome of Decumbent Corydalis is alkaloid." Chinese Pharmacopoeia " 2010 editions one is also using the quantitative and qualitative analysis index of alkaloid as Rhizome of Decumbent Corydalis.
Total alkaloid of corydalis rhizome mainly comprises protopine, tetrahydropalmatine, summer rather alkali, Bicuculline, palmatine hydrochloride.Protopine is a kind of isoquinoline alkaloid, has the activity of cardiovascular aspect as reduced hypertension, anti-heart disorder; Also there is the multiple pharmacologically actives such as platelet aggregation-against, anti-immunity, anti-oxidative damage.Tetrahydropalmatine equally also has the multiple pharmacological effect such as antitumor, platelet aggregation-against, anti-arrhythmia, analgesia.The pharmacologically active that Rhizome of Decumbent Corydalis alkaloid is numerous causes to be paid close attention to widely, and therefore their separation purification method is not only the needs of research pharmacological action further, simultaneously also for clinical study provides material base.Make a general survey of existing Rhizome of Decumbent Corydalis alkaloid monomer separation purification method, mostly adopt column chromatography, crystallization and prepare the method for liquid phase, technique is loaded down with trivial details, the rate of recovery is low, cost is more high.
FCPC technology and high speed centrifugation distribution chromatography, it utilizes the partition ratio of material to be separated in immiscible two phase liquid different, through being similar to the process of continuous extraction, is separated material.It and HSCCC are slightly different, and HSCCC belongs to hydrodynamic-balanced system, the chromatographic column turned to by teflon hose except rotating around centrifugal shaft, also around rotating the gravity field changed from axle; FCPC belongs to hydrostatic equilibrium system, and chromatographic column is made up of a series of chamber be engraved on disk, produces constant centrifuge field thus realize the reservation to stationary phase by single-shaft-rotation.The nineties in 20th century, Ito has invented pH-zone refining countercurrent, and the method is very suitable for separating organic acid and organic bases compounds.This method is equally also applicable in FCPC, and this method has following characteristics: solute is highly concentrated in rectangle peak, and overlapping less between different rectangle peak, and the basis of separation has different hydrophobicitys and pKa value based on different solute.Compared with traditional adverse current chromatogram method, pH-zone refining countercurrent has many important advantages, as substantially increased the applied sample amount of sample, is more than 10 times of usual way; Large component high enrichment, the high enrichment of minor component.The method is own through being widely used in the material of separating natural product and synthetic at present, especially alkaloid and organic acid.
Summary of the invention
Technical problem to be solved by this invention is the shortcomings such as the preparation method's cost in order to overcome lifeless matter alkali monomer in summer in prior art is higher, separation efficiency is low, the rate of recovery is low, product purity is low, preparation amount is little and provides a kind of preparation method of Rhizome of Decumbent Corydalis alkaloid monomer, this preparation method can prepare each alkaloid monomer on a large scale, prepares the highlyest can reach a gram level amount; And obtained monomer purity is high, reaches more than 98%; Cost is low, separation efficiency is high, the rate of recovery is high, product purity is high, preparation amount is larger.
The invention provides a kind of preparation method of Rhizome of Decumbent Corydalis alkaloid monomer, it comprises the following steps: adopt high speed centrifugation partition chromatography to carry out separation and purification to Rhizoma Corydalis Decumbentis extract, obtain Rhizome of Decumbent Corydalis alkaloid monomer; Described Rhizome of Decumbent Corydalis alkaloid monomer is protopine, tetrahydropalmatine or summer rather alkali;
Wherein, the stationary phase that described high speed centrifugation partition chromatography uses and moving phase are obtained by following method: sherwood oil, ethyl acetate, second alcohol and water are mixed, after stratification, by upper phase, lower phase solution separately, upper phase solution is mixed with triethylamine and obtains stationary phase, namely lower phase solution and mixed in hydrochloric acid are evenly obtained moving phase; The volume ratio of described sherwood oil, ethyl acetate, second alcohol and water is (1 ~ 5): (1 ~ 10): (1 ~ 5): (1 ~ 10); In described stationary phase, the volumetric molar concentration of triethylamine is 5nM ~ 25nM; In described moving phase, the volumetric molar concentration of HCl is 5nM ~ 25nM.
In the present invention, described Rhizoma Corydalis Decumbentis extract can be the Rhizoma Corydalis Decumbentis extract that this area routine uses; In described Rhizoma Corydalis Decumbentis extract, the mass percentage of total alkaloid of corydalis rhizome is 50% ~ 99%, and described mass percentage refers to that in Rhizoma Corydalis Decumbentis extract, the quality of total alkaloids accounts for the per-cent of Rhizoma Corydalis Decumbentis extract quality.In total alkaloid of corydalis rhizome, the proportion of composing of several main alkaloid is as follows: protopine 15% ~ 35%, tetrahydropalmatine 10% ~ 25%, Bicuculline 5% ~ 15%, palmatine hydrochloride 0.05% ~ 0.2%, corydalis that bright 5% ~ 15%, and the summer is alkali 15% ~ 30%, related alkaloids and extract 10% ~ 25% rather; Preferred 0.5g ~ the 4g of described Rhizoma Corydalis Decumbentis extract, further preferred 0.8g ~ 1.6g.The preferred Rhizome of Decumbent Corydalis supercritical extract of described Rhizoma Corydalis Decumbentis extract; Supercritical extraction method preparation (see the application's reference example 1) of the Rhizome of Decumbent Corydalis that described Rhizome of Decumbent Corydalis supercritical extract is recorded according to CN101058576A.
In the present invention, the volume ratio preferred (3 ~ 5) of described sherwood oil, ethyl acetate, second alcohol and water: 7:(3 ~ 5): 7.
In the present invention, the preferred 5nM ~ 20nM of the volumetric molar concentration of triethylamine in described stationary phase, further preferred 10nM.
In the present invention, the preferred concentrated hydrochloric acid of described hydrochloric acid.
In the present invention, the preferred 5nM ~ 10nM of the volumetric molar concentration of HCl in described moving phase.
In the present invention, in the preparation process of described stationary phase and moving phase, the time of described stratification can be the stratification time of this area routine, preferably 2 ~ 12h, further preferred 4 ~ 12h.
In the present invention, described high speed centrifugation partition chromatography is carried out in high speed centrifugation distribution chromatography instrument; Described high speed centrifugation distribution chromatography instrument is the high speed centrifugation distribution chromatography instrument of this area routine, and the model that preferred French RousseletRobatelKromaton company produces is the high speed centrifugation distribution chromatography instrument of FCPCA200.The operation parameter condition of described high speed centrifugation distribution chromatography instrument can be the Parameter Conditions of this area routine.
In the present invention, described high speed centrifugation partition chromatography preferably comprises the steps:
(1) Rhizoma Corydalis Decumbentis extract is dissolved in upper phase solution or lower phase solution, obtains sample introduction solution;
(2) rotary drum of high speed centrifugation distribution chromatography instrument is filled with stationary phase, after being full of stationary phase in rotary drum, regulate stationary phase flow velocity, and by described sample introduction solution sample introduction, under 1000rpm ~ 1600rpm rotating speed, pump into moving phase, collect the corresponding moving phase flowed out.
In step (1), the mass volume ratio preferably 0.1 ~ 0.8g/mL of the described Rhizoma Corydalis Decumbentis extract described in sample introduction liquid and described stationary phase or moving phase, further preferred 0.32g/mL.
In step (2), before being full of stationary phase in rotary drum, the preferred 15mL/min of stationary phase flow velocity.
In step (2), described is full of for there being stationary phase to flow out rotary drum.
In step (2), after being full of stationary phase in rotary drum, stationary phase flow velocity preferably 2 ~ 6mL/min, further preferred 4mL/min.
In step (2), the flow velocity of described sample introduction, flow rate of mobile phase, the stationary phase flow velocity after being full of stationary phase with in rotary drum is identical.
In step (2), the preferred 1200rpm/min of described rotating speed.
In step (2), described collection is determined according to FCPC spectrogram, and FCPC adopts ultraviolet monitor function, and described ultraviolet wavelength is 282nm.
In the present invention, agents useful for same and raw material are all commercially.
Positive progressive effect of the present invention is: the present invention can prepare each alkaloid monomer on a large scale, prepares the highlyest can reach a gram level amount; And obtained monomer purity is high, reaches more than 98%; Separation efficiency is high, the rate of recovery is high, product purity is high, preparation amount is larger.
Accompanying drawing explanation
Fig. 1 is that Rhizome of Decumbent Corydalis supercritical extract FCPC schemes.
Fig. 2 is that Rhizome of Decumbent Corydalis supercritical extract HPLC schemes.
Embodiment
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or selects according to catalogue.
Instrument: supercritical extraction instrument HA121-50-01-C (the overcritical company limited in Huaan, Nantong), AgilentHP1260 type high performance liquid chromatograph, quaternary gradient pump, Hpcheme chromatographic working station.MSU224S-100-Duz balance (Sartorius); High speed centrifugation distribution chromatography (FCPC) (Kromaton, RousseletRobatel); SHIMDZULC-10AD pump; HD-21-2 UV-detector (Shanghai Jia Peng Science and Technology Ltd.); SHIMDZU-7725i sampler.
The preparation of reference example 1 Rhizome of Decumbent Corydalis supercritical extract
Get the dry tuber of Rhizome of Decumbent Corydalis, be ground into 20 order ~ 100 object medicinal powders; Get medicinal powder a certain amount of, add ammoniacal liquor, mix to pH8 ~ 10; Medicinal material is dropped in extraction kettle, extraction temperature 40 DEG C ~ 75 DEG C, extraction-container I temperature 30 DEG C ~ 70 DEG C, during extraction-container II temperature 30 DEG C ~ 70 DEG C, extract under extracting pressure 30MPa ~ 50MPa, add ethanol continuously as entrainment agent in extraction process, adding flow velocity is 260mL/h; When extraction is after 2 ~ 3 hours, stop extraction, analytically still I obtains total alkaloids extract, is Rhizoma corydalis decumbentis extract.All examples all adopt same batch sample to carry out below, and wherein Bicuculline content is 7.81%, the summer rather alkali content be 19.21%, protopine content is 5.43%, and Content determination of dl-tetrahydropalmatine is 18.12%, and palmatine content is 0.12%.
Embodiment 1
Selected sherwood oil: ethyl acetate: ethanol: water=3:7:3:7 is system is separation system, adds in separating funnel respectively in its ratio by all kinds of SOLVENTS, and each solvent of concuss fully mixes, and after balance, gets upper and lower phase respectively.In upper phase solution, add triethylamine, be made into the stationary phase containing triethylamine 5nM, in lower phase solution, add hydrochloric acid, be made into the moving phase of hydrochloric 5nM, Rhizome of Decumbent Corydalis supercritical extract 1.6g to be dissolved on 5ml in phase solution; Under Descending pattern, in FCPC rotary drum, stationary phase is pumped into 15ml/min flow velocity, after being full of stationary phase in post, regulate flow velocity to 4ml/min and sample introduction, then start to pump into moving phase with 4ml/min under 1200rpm rotating speed, and receiving the corresponding elutriant of each alkali according to detector collection of illustrative plates, namely evaporate to dryness elutriant obtains corresponding alkaloid.Protopine 62mg, yield 71%, purity 98.1%; Summer is alkali 274mg rather, yield 89%, purity 98.5%; Tetrahydropalmatine 264mg, yield 91%, purity 98.8%.
Embodiment 2
Selected sherwood oil: ethyl acetate: ethanol: water=4:7:4:7 is system is separation system, adds in separating funnel respectively in its ratio by all kinds of SOLVENTS, and each solvent of concuss fully mixes, after balance, gets phase solution and lower phase solution respectively.In upper phase solution, add triethylamine, be made into the stationary phase containing triethylamine 20nM, in lower phase solution, add hydrochloric acid, be made into the moving phase of hydrochloric 10nM, Rhizome of Decumbent Corydalis supercritical extract 0.8g to be dissolved on 5mL in phase solution; Under Descending pattern, in FCPC rotary drum, stationary phase is pumped into 15ml/min flow velocity, after being full of stationary phase in post, regulate flow velocity to 4ml/min and sample introduction, then start to pump into moving phase with 4ml/min under 1200rpm rotating speed, and receiving the corresponding elutriant of each alkali according to detector collection of illustrative plates, namely evaporate to dryness elutriant obtains corresponding alkaloid.Protopine 30mg, yield 69%, purity 98.2%; Summer is alkali 143mg rather, yield 93%, purity 98.6%; Tetrahydropalmatine 138mg, yield 95%, purity 98.4%.
Embodiment 3
Selected sherwood oil: ethyl acetate: ethanol: water=5:7:5:7 is system is separation system, adds in separating funnel respectively in its ratio by all kinds of SOLVENTS, and each solvent of concuss fully mixes, after balance, gets phase solution and lower phase solution respectively.In upper phase solution, add triethylamine, be made into the stationary phase containing triethylamine 10nM, in lower phase solution, add hydrochloric acid, be made into the moving phase of hydrochloric 10nM, Rhizome of Decumbent Corydalis supercritical extract 1.6g to be dissolved under 10mL in phase solution; Under Descending pattern, in FCPC rotary drum, stationary phase is pumped into 15ml/min flow velocity, after being full of stationary phase in post, regulate flow velocity to 4ml/min and sample introduction, then start to pump into moving phase with 4ml/min under 1200rpm rotating speed, and receiving the corresponding elutriant of each alkali according to detector collection of illustrative plates, namely evaporate to dryness elutriant obtains corresponding alkaloid.Protopine 66mg, yield 76%, HPLC purity 98.3%; Summer is alkali 287mg rather, yield 93%, HPLC purity 98.6%; Tetrahydropalmatine 268mg, yield 92%, HPLC purity 98.9%.Rhizome of Decumbent Corydalis supercritical extract FCPC figure is shown in that Fig. 1 (wherein, collect at 76 ~ 80min by protopine; Summer rather alkali collect at 83 ~ 98min; Tetrahydropalmatine is collected at 102 ~ 110min).Protopine
1h-NMR data in table 2, protopine
13c-NMR data in table 3, summer rather alkali
1h-NMR data in table 4, summer rather alkali
13c-NMR data in table 5, tetrahydropalmatine
1h-NMR data are in table 6.
Detection method
Quantitatively take each alkaloid reference substance, dissolve with mobile phase A (%): B (%)=88:12 (described % represents volume percent), prepare the Bicuculline 0.0532mg/mL that each concentration is following, summer is alkali 0.1202mg/mL rather, protopine 0.0764mg/mL, tetrahydropalmatine 0.1254mg/mL, palmatine 0.0446mg/mL reference substance solution.Rhizome of Decumbent Corydalis supercritical extract HPLC (high performance liquid chromatography) spectrogram see Fig. 2 (wherein, the summer rather alkali go out peak at 35min; Protopine goes out peak at 45min; Tetrahydropalmatine goes out peak at 67min).
Chromatographic condition is as follows: chromatographic column: Discovery (5 μm × 250mm).Testing conditions is A phase: 8mL triethylamine, and 30mL Glacial acetic acid to add in water constant volume to 1000mL; B phase: methyl alcohol: acetonitrile=1:4; Elution flow Phase Proportion is in table 1.Flow velocity: 1mL/min.Determined wavelength: 282nm.
Table 1 mobile phase ratio
| Time (min) | A(％) | B(％) |
| 0 | 88 | 12 |
| 50 | 88 | 12 |
| 75 | 75 | 25 |
Table 2 protopine
1h-NMR data inCDCl
3(400MHz)
| δH | |
| C 1,-H | 6.93(s) |
| C 4,-H | 6.67(s) |
| C 5,6-H 2 | 2.55(br,s) |
| C 8-H 2 | 3.55(s) |
| C 11-H | 6.70(s) |
| C 12-H | 6.69(s) |
| C 13-H 2 | 3.81(s) |
| N-CH 3 | 1.94(s) |
| C 2-O-CH 2-O-C 3 | 5.95(s) |
| C 9-O-CH 2-O-C 10 | 5.97(s) |
Table 3 protopine
13c-NMR data inCDCl
3(100MHz)
| δC | |
| C 1 | 108.128 |
| C 2 | 146.350 |
| C 3 | 148.019 |
| C 4 | 110.478 |
| C 5 | 31.815 |
| C 6 | 57.853 |
| C 8 | 50.851 |
| C 9 | 146.010 |
| C 10 | 145.889 |
| C 11 | 106.716 |
| C 12 | 125.106 |
| C 13 | 46.540 |
| C 14 | 195.011 |
| C 1a | 136.222 |
| C 12a | 129.014 |
| C 14a | 195.011 |
| C 8a | 117.976 |
| C 2-O-CH 2-O-C 3 | 101.189 |
| C 9-O-CH 2-O-C 1 | 100.84 |
| N-CH 3 | 41.454 |
Table 4 summer rather alkali
1h-NMR data inCDCl
3(400MHz)
| δ H | |
| C 1-H | 2.50(s) |
| C 9-H | 3.88(d, |
| C 5-H | 5.40(d, |
| C 8-H | 6.58(s) |
| C 6.7-OCH 2O- | 6.78(s) |
| C 4’.5’-OCH 2O | 5.98(s),5.96 |
| C 3’-H | 6.02(s),6.00 |
| C 2’-H | 6.53(d, |
| C 7’-H | 5.64(d, |
| N-CH 3 | 6.33(s) |
Table 5 summer rather alkali
13c-NMR data inCDCl
3(100MHz)
| δ C | δ C | ||
| NCH 3 | 44.1 | C 9 | 87.0 |
| C 1 | 65.1 | C 6’ | 114.7 |
| C 3 | 46.1 | C 5’ | 141.8 |
| C 4 | 22.5 | C 4’ | 148.3 |
| C 4a | 124.2 | C 3’ | 108.4 |
| C 5 | 107.6 | C 2’ | 114.5 |
| C 6 | 146.7 | C 1’ | 133.2 |
| C 7 | 146.5 | C 6.7-OCH2O- | 100.9 |
| C 8 | 108.7 | C 4.5-OCH2O- | 101.7 |
| C 8a | 128.7 | C 7’ | 98.0 |
Table 6 tetrahydropalmatine
1h-NMR data inCDCl
3(400MHz)
| δ H | |
| C 1-H | 6.62(s,1H) |
| C 4-H | 6.73(s,1H) |
| C 5,6-H | 2.84～3.29(m,4H) |
| C 8-H | 4.26(d,J＝16Hz,1H) |
| C 11-H | 6.79(d,J＝8.5Hz,1H) |
| C 12-H | 6.88(d,J＝8.5Hz,1H) |
| C 13-H | 2.67(1H)，2.70(1H) |
| C 14-H | 3.57(d,J＝13Hz,1H) |
| C 2,3,9,10-O-C | 3.89(s)，3.87(s)，3.86(s)， |
Claims (10)
1. a preparation method for Rhizome of Decumbent Corydalis alkaloid monomer, is characterized in that, comprises the following steps: adopt high speed centrifugation partition chromatography to carry out separation and purification to Rhizoma Corydalis Decumbentis extract, obtain Rhizome of Decumbent Corydalis alkaloid monomer; Described Rhizome of Decumbent Corydalis alkaloid monomer is protopine, tetrahydropalmatine or summer rather alkali;
Wherein, the stationary phase that described high speed centrifugation partition chromatography uses and moving phase are obtained by following method: sherwood oil, ethyl acetate, second alcohol and water are mixed, after stratification, by upper phase, lower phase solution separately, upper phase solution is mixed with triethylamine and obtains stationary phase, namely lower phase solution and mixed in hydrochloric acid are evenly obtained moving phase; The volume ratio of described sherwood oil, ethyl acetate, second alcohol and water is (1 ~ 5): (1 ~ 10): (1 ~ 5): (1 ~ 10); In described stationary phase, the volumetric molar concentration of triethylamine is 5nM ~ 25nM; In described moving phase, the volumetric molar concentration of HCl is 5nM ~ 25nM.
2. preparation method as claimed in claim 1, it is characterized in that, in described Rhizoma Corydalis Decumbentis extract, the mass percentage of total alkaloid of corydalis rhizome is 50% ~ 99%;
And/or described Rhizoma Corydalis Decumbentis extract is 0.5g ~ 4g;
And/or described Rhizoma Corydalis Decumbentis extract is Rhizome of Decumbent Corydalis supercritical extract.
3. preparation method as claimed in claim 1, is characterized in that, in the preparation process of described stationary phase and moving phase, the time of described stratification is 2 ~ 12h;
And/or the volume ratio of described sherwood oil, ethyl acetate, second alcohol and water is (3 ~ 5): 7:(3 ~ 5): 7;
And/or the volumetric molar concentration of triethylamine is 5nM ~ 20nM in described stationary phase;
And/or described hydrochloric acid is concentrated hydrochloric acid;
And/or the volumetric molar concentration of HCl is 5nM ~ 10nM in described moving phase.
4. preparation method as claimed in claim 3, is characterized in that, in the preparation process of described stationary phase and moving phase, the time of described stratification is 4 ~ 12h;
And/or the volumetric molar concentration of triethylamine is 10nM in described stationary phase.
5. preparation method as claimed in claim 1, it is characterized in that, described high speed centrifugation partition chromatography is carried out in high speed centrifugation distribution chromatography instrument.
6. preparation method as claimed in claim 5, is characterized in that, the high speed centrifugation distribution chromatography instrument of described high speed centrifugation distribution chromatography instrument to be model that French RousseletRobatelKromaton company produces be FCPCA200.
7. preparation method as claimed in claim 1, it is characterized in that, described high speed centrifugation partition chromatography comprises the steps:
(1) Rhizoma Corydalis Decumbentis extract is dissolved in upper phase solution or lower phase solution, obtains sample introduction solution;
(2) rotary drum of high speed centrifugation distribution chromatography instrument is filled with stationary phase, after being full of stationary phase in rotary drum, regulate stationary phase flow velocity, and by described sample introduction solution sample introduction, under 1000rpm ~ 1600rpm rotating speed, pump into moving phase, collect the corresponding moving phase flowed out.
8. preparation method as claimed in claim 7, it is characterized in that, in step (1), the mass volume ratio of described Rhizoma Corydalis Decumbentis extract and described stationary phase or moving phase is 0.1 ~ 0.8g/mL;
And/or in step (2), described high speed centrifugation distribution chromatography instrument is the model that French RousseletRobatelKromaton company produces is the high speed centrifugation distribution chromatography instrument of FCPCA200;
And/or in step (2), before being full of stationary phase in rotary drum, stationary phase flow velocity is 15mL/min;
And/or in step (2), after being full of stationary phase in rotary drum, stationary phase flow velocity is 2 ~ 6mL/min;
And/or, in step (2), the flow velocity of described sample introduction and flow rate of mobile phase, the stationary phase flow velocity after being full of stationary phase with in rotary drum is identical;
And/or in step (2), described rotating speed is 1200rpm.
9. preparation method as claimed in claim 8, is characterized in that,
In step (1), the mass volume ratio of described Rhizoma Corydalis Decumbentis extract and described stationary phase or moving phase is 0.32g/mL;
And/or in step (2), after being full of stationary phase in rotary drum, stationary phase flow velocity is 4mL/min.
10. preparation method as claimed in claim 7, it is characterized in that, in step (2), described collection is determined according to FCPC spectrogram.
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