CN1583725A - Preparation of liensinine, isoliensinine and methylliensinine extracted from lotus seed - Google Patents

Preparation of liensinine, isoliensinine and methylliensinine extracted from lotus seed Download PDF

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CN1583725A
CN1583725A CN 200410025198 CN200410025198A CN1583725A CN 1583725 A CN1583725 A CN 1583725A CN 200410025198 CN200410025198 CN 200410025198 CN 200410025198 A CN200410025198 A CN 200410025198A CN 1583725 A CN1583725 A CN 1583725A
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plumula nelumbinis
liensinine
isoliensinine
neferine
phase
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CN1286816C (en
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潘远江
吴世华
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Zhejiang University ZJU
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Zhejiang University ZJU
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Abstract

A process for preparing liensinine, iso liensinine and methyl liensinine from the core of lotus seed by counterflow chromatography method features that its separating solvent is the mixture of two or more in n-alkanes, halohydrocarbon, emtrol, lipone, fatty ester, ether and water.

Description

From Plumula Nelumbinis, separate the method for preparing liensinine, Isoliensinine and Neferine
Technical field
The present invention relates to a kind of chromatography that adopts and from Plumula Nelumbinis, separate the method for preparing high-purity monomer liensinine, Isoliensinine, Neferine.
Background technology
Liensinine, Isoliensinine, Neferine are the main alkaloid of Plumula Nelumbinis.Plumula Nelumbinis is the plumule of Nymphaeceae nelumbium seed, is one of important Chinese medicine of China, has functions such as clearing away the heart-fire, hypotensive, cardiac stimulant.A large amount of effective constituents that studies show that Plumula Nelumbinis are these three kinds of alkaloids.Because these three kinds of alkaloids, separate the important content that the high-purity compound monomer of preparation becomes research in the outstanding curative effect of aspects such as cardiovascular and cerebrovascular.Yet these three kinds of alkaloidal chemical property of phenol are unstable, easily oxidation, nor easily form crystallization.Therefore, when adopting traditional isolation technique such as thin-layer chromatography, column chromatography and high performance liquid chromatography etc., usually produce hangover, pollution, loss because of the absorption of solid carrier, the result is difficult to obtain the compound monomer of higher degree.
Chinese patent CN1117735C discloses a kind of method of extracting purifying Isoliensinine and liensinine from Plumula Nelumbinis, and this method is that complex operation step is difficult to large-scale production with fractional crystallization behind the embryo nelumbinis alkaloid total alkali salify.Therefore, press for a kind of efficiently, separation method fast, liensinine, Isoliensinine, Neferine are prepared so that higher yield is separated from the Plumula Nelumbinis alkaloid crude product.
Summary of the invention
The purpose of this invention is to provide a kind of low cost, high-recovery, be suitable for the extensive method for preparing liensinine, Isoliensinine and Neferine of from Plumula Nelumbinis, separating.
The method prepare liensinine, Isoliensinine and Neferine of from Plumula Nelumbinis, separating of the present invention, employing be the adverse current chromatogram method, may further comprise the steps:
1) with Plumula Nelumbinis powder with organic solvent lixiviate repeatedly, extracting solution is merged, is condensed into medicinal extract, gained medicinal extract is lower than the dissolving of 20% aqueous acid with concentration, filters, filtrate transfers PH to alkalescence, separates out throw out, obtains the Plumula Nelumbinis alkaloid crude product; Perhaps Plumula Nelumbinis powder directly is lower than 20% aqueous acid lixiviate with concentration, filters, filtrate transfers PH to alkalescence, separates out throw out, obtains the Plumula Nelumbinis alkaloid crude product;
2) will be used for isolating solvent and pour skimmer into, and form immiscible two-phase solvent up and down, the biphase volume ratio is 0.5-2.0 up and down;
3) the Plumula Nelumbinis alkaloid crude product is dissolved in step 2) two-phase solvent in, obtain the Plumula Nelumbinis alkaloid sample solution;
4) with step 2) one in the two-phase solvent inject in the counter current chromatograph separator column mutually as stationary phase, the Plumula Nelumbinis alkaloid crude product solution of step 3) gained is injected the injection annulus of counter current chromatograph, open counter current chromatograph, another of injection two-phase solvent made mobile phase mutually, Plumula Nelumbinis alkaloid just carries out successive and distributes in counter current chromatograph, by detection at thread detector, collect, merge effluent, obtain liensinine, Isoliensinine and the Neferine of purifying respectively.
Above-mentioned steps 1) said organic solvent can be normal paraffin, halohydrocarbon, Fatty Alcohol(C12-C14 and C12-C18), aliphatic ketone, fatty ester or ether.
Above-mentioned steps 2) saidly be used for two or more the mixture that isolating solvent can be normal paraffin, halohydrocarbon, Fatty Alcohol(C12-C14 and C12-C18), aliphatic ketone, fatty ester, ethers or water, these solvents have partition ratio preferably to liensinine, Isoliensinine and Neferine.
The adverse current chromatogram method that the present invention adopts is the liquid liquid distribution chromatography method that a kind of successive need not any solid support, the problems such as absorption, loss and sex change that it has avoided solid state adhesion body or carrier to cause, have and to guarantee higher peak type resolution, fractional dose is big, sample free of losses, rate of recovery height, isolating environment relax, save characteristics such as solvent, can directly separate slightly get sample in a large number product or synthetic mixture, separating resulting purity can reach more than 95%.And used counter current chromatograph is a General Instrument, its more efficient liquid chromatograph far is a simple cheap, its solvent is prepared by usual vehicle, and therefore low price is a kind of method of separating liensinine, Isoliensinine and Neferine from Plumula Nelumbinis extract of economy.
Description of drawings
Fig. 1 is the high-efficient liquid phase analysis figure of Plumula Nelumbinis alkaloid crude product among the embodiment 1;
Fig. 2 is the high-efficient liquid phase analysis figure of the compound of the isolating adverse current chromatogram of Plumula Nelumbinis alkaloid and purifying among the embodiment 1;
Fig. 3 is the chemical structural drawing of liensinine, Isoliensinine and Neferine among the embodiment 1;
Fig. 4 is the high-efficient liquid phase analysis figure of the compound of the isolating adverse current chromatogram of Plumula Nelumbinis alkaloid and purifying among the embodiment 2.
Embodiment
The invention will be further described below in conjunction with embodiment and accompanying drawing.
Embodiment 1
1. the Plumula Nelumbinis powder that 12kg is done was with 95% ethanol 10L room temperature cold soaking 48 hours, and suction filtration goes out solvent, adds 95% ethanol of 10L in filter residue again, repeats above operation three times, and the extracting solution merging is become medicinal extract with the rotatory evaporator concentrating under reduced pressure, and thickening temperature is 40 ℃.Gained medicinal extract is with 1.5% the dissolving with hydrochloric acid of 4L, suction filtration, and filtrate is separated out faint yellow precipitation with 10% ammoniacal liquor accent PH to 8.5.The faint yellow precipitation that collection is separated out gets 85g Plumula Nelumbinis alkaloid crude product.Efficient liquid phase chromatographic analysis (as Fig. 1,1 is liensinine among the figure, and 2 is Isoliensinine, and 3 are Neferine) shows that the content of three kinds of main components is respectively 15.4%, 15.9%, 47.6%.
2. separate the Plumula Nelumbinis alkaloid extract with counter current chromatograph.With sherwood oil, ethyl acetate, tetracol phenixin, chloroform, first alcohol and water, be 1: 1: 4 by volume: 4: 6: 1 preparation two-phase solvents, standing demix in liquid separatnig container is told phase up and down.Plumula Nelumbinis alkaloid crude product 1102mg is dissolved in that 9ml goes up mutually and under the 9ml mutually in the formation sample solution.Adverse current chromatogram column internal diameter 4mm, column volume are 1600ml.Phase mutually below fixes, be mobile phase mutually down, after filling with stationary phase in treating in the counter current chromatograph separator column, sample solution is injected the injection annulus of counter current chromatograph, open counter current chromatograph, rotating speed is 450 to change, flow velocity with 5ml/min injects mobile phase, UV-detector monitoring flow point by online detects wavelength 280nm, collects, merges effluent, obtain liensinine, Isoliensinine and Neferine respectively, its purity is respectively 97.45%, 95.32%, and 98.43%, see Fig. 2,1 is liensinine among the figure, and 2 is Isoliensinine, and 3 is Neferine.Separate the compound structure that obtains as shown in Figure 3 through modern wave spectrum test proof through adverse current chromatogram, 1 is liensinine among the figure, and 2 is Isoliensinine, and 3 is Neferine.Table 1-table 6 has been listed a peacekeeping two dimensional NMR spectral data of liensinine, Isoliensinine and the Neferine of gained respectively.
Table 1. liensinine 1H-NMR and 1H- 1H COSY spectral data
The atom sequence number Chemical shift δ, ppm Multiplicity ???? 1H- 1H?COSY
????1 ????3.50 ??1H，d，J＝8.30Hz ????H-9
????2-CH 3 ????2.58 ??3H，s
????3ax ????2.74 ??1H，m ????H-3eq，H-4
????3eq ????3.19 ??1H，m ????H-3ax，H-4
????4 ????2.84 ??2H，m ????H-4ax，H-3
????5 ????6.67 ??1H，s
????6-OCH 3 ????3.88 ??3H，s
????8 ????6.45 ??1H，s
????9ax ????2.69 ??1H，m ????H-9eq，H-1
????9eq ????3.22 ??1H，m ????H-1，H-9ax
????11 ????6.98 ??1H，d，J＝8.15 ????H-12
????12 ????6.79 ??1H，d，J＝8.15 ????H-11
????14 ????6.79 ??1H，d，J＝8.15 ????H-15
????15 ????6.98 ??1H，d，J＝8.15 ????H-14
????1′ ????3.69 ????1H，d，J＝9.00Hz ????H-9′
????2′-CH 3 ????2.54 ????3H，s
????3′ax ????3.00 ????1H，m ????H-3′eq，H-4′
????3′-eq ????3.46 ????1H，m ????H-4′，H-3′ax
????4′ax ????2.59 ????1H，m ????H-4′eq，H-3′
????4′eq ????3.01 ????1H，m ????H-3′，H-4′ax
????5′ ????6.58 ????1H，s
????6′-OCH 3 ????3.84 ????3H，s
????7′-OCH 3 ????3.43 ????3H，s
????8′ ????5.71 ????1H，s
????9′ax ????2.62 ????1H，m ????H-9′eq，H-1′
????9′eq ????3.13 ????1H，m ????H-9′ax，H-1′
????11′ ????6.81 ????1H，s
????14′ ????6.72 ????1H，d，J＝8.15 ????H-15′
????15′ ????6.39 ????1H，d，J＝8.15 ????H-14′
Table 2. liensinine 13C-NMR and 2D NMR spectral data
The atom sequence number ? 13C-NMR DEPT a ????HMQC ????HMBC
????1 ??65.604 ??CH ????3.5 ????H9，2-CH 3
????2-CH 3 ??42.709 ??CH 3 ????2.58 ????H-1，H-3
????3 ??48.143 ??CH 2 ????3.19，2.74 ????H-4，2-CH 3
????4 ??26.57 ??CH 2 ????2.84 ????H-3，H-5
????4a ??129.773 ??q b ????H-8，H-4
????5 ??112.17 ??CH ????6.67
????6 ??148.223 ??q ????H-5，H-8，6-OCH 3
????6-OCH 3 ??55.985 ??CH 3 ????3.88
????7 ??143.378 ??q ????H-5，H-8
????8 ??118.027 ??CH ????6.45 ????H-1
????8a ??130.671 ??q ????H-1，H-8
????9 ??40.087 ??CH 2 ????3.22，2.69 ????H-1，H-11，H-15
????10 ??130.725 ??q ????H-9，H-11，H-15
????11 ??130.992 ??CH ????6.98 ????H-12，H-15
????12 ??116.921 ??CH ????6.79 ????H-11，H-14
????13 ??155.755 ??q ????H-11，H-15，H-12，H-14
????14 ??116.921 ??CH ????6.79 ????H-12，H-15
????15 ??130.992 ??CH ????6.98 ????H-11，H-14
????1′ ??64.655 ??CH ????3.69 ????H-9′，2′-CH 3
????2′-CH 3 ??40.435 ??CH 3 ????2.54 ????H-3′，H-1′
????3′ ??44.237 ??CH 2 ????3.46，3.00 ????H-4′，2′-CH 3
????4′ ??21.705 ??CH 2 ????3.01，2.59 ????H-5′
????4′a ??123.427 ??q ????H-4′，H-3′，H-8′
????5′ ??111.349 ??CH ????6.58
????6′ ??147.75 ??q ????H-5′，H-8′，6′-OCH 3
????6′-OCH 3 ??55.826 ??CH 3 ????3.84
????7′ ??146.307 ??q ????H-5′，H-8′，7′-OCH 3
????7′-OCH 3 ??55.423 ??CH 3 ????3.43
????8′ ??111.474 ??CH ????5.71 ????H-1′
????8′a ??126.654 ??q ????H-5′，H-1′，H-9′
????9′ ??42.546 ??CH 2 ????3.13，2.62 ????H-1′，H-11′，H-15′
????10′ ??130.725 ??q ????H-11′，H-14′，H-9′
????11′ ??121.827 ??CH ????6.81 ????H-15′，H-9′
????12′ ??144.101 ??q ????H-11′，H-14′
????13′ ??146.462 ??q ????H-14′，H-15′，H-11′
????14′ ??115.377 ??CH ????6.72
????15′ ??127.496 ??CH ????6.39 ????H-11′，H-9′
aDEPT 135. bQuaternary carbon.
Table 3. Isoliensinine 1H-NMR and 1H- 1H COSY spectral data
The atom sequence number Chemical shift δ, ppm Multiplicity ?? 1H- 1H?COSY
????1 ????3.66 ????1H，m ????H-9
??2-CH 3 ????2.39 ????3H，s
????3ax ????2.64 ????1H，m ????H-3eq，H-4
????3eq ????3.08 ????1H，m ????H-4，H-3ax
????4ax ????2.47 ????1H，m ????H-4eq，H-3
????4eq ????2.70 ????1H，m ????H-4ax，H-3
????5 ????6.47 ????1H，s
????6-OCH 3 ????3.81 ????3H，s
????8 ????6.34 ????1H，s
????9ax ????2.77 ????1H，m ????H-9eq，H-1
????9eq ????3.03 ????1H，m ????H-9ax，H-1
????11 ????6.90 ????1H，d，J＝8.42 ????H-12
????12 ????6.70 ????1H，d，J＝8.42 ????H-11
????13-OCH 3 ????3.73 ????3H，s
????14 ????6.70 ????1H，d，J＝8.42 ????H-15
????15 ????6.90 ????1H，d，J＝8.42 ????H-14
????1′ ????3.57 ????1H，m ????H-9′
????2′-CH 3 ????2.51 ????3H，s
????3′ax ????2.78 ????1H，m ????H-3′eq，H-4′
????3′eq ????3.16 ????1H，m ????H-4′，H-3′ax
????4′ax ????2.66 ????1H，m ????H-4′eq，H-3′
????4′eq ????2.87 ????1H，m ????H-4′ax，H-3′
????5′ ????6.64 ????1H，s
????6′-OCH 3 ????3.79 ????3H，s
????8′ ????6.30 ????1H，s
????9′ax ????2.80 ????1H，m ????H-9′eq，H-1′
????9′eq ????2.93 ????1H，m ????H-9′ax，H-1′
????11′ ????6.46 ????1H，s
????14′ ????6.81 ????1H，d，J＝8.16 ????H-15′
????15′ ????6.72 ????1H，J＝8.16 ????H-14′
Table 4. Isoliensinine 13C-NMR and 2D NMR spectral data
The atom sequence number ??? 13C-NMR DEPT a ??HMQC ????HMBC
????1 ????64.569 ??CH ????3.66 ????H-8，2-CH 3，H-9
????2-CH 3 ????42.692 ??CH 3 ????2.39
????3 ????47.468 ??CH 2 ????3.08，2.64 ????2-CH 3
????4 ????25.562 ??CH 2 ????2.70，2.47 ????H-5，H-3
????4a ????130.034 ??qb ????H-5
????5 ????110.640 ??CH ????6.47
????6 ????145.254 ??q ????H-5，6-OCH 3
????6-OCH 3 ????55.982 ??CH 3 ????3.81
????7 ????143.568 ??q ????H-5，H-8
????8 ????113.869 ??CH ????6.34
????8a ????125.601 ??q ????H-8，H-4
????9 ????39.339 ??CH 2 ????3.03，2.77 ????H-11，H-15
????10 ????131.436 ??q ????H-12，H-14
????11 ????130.503 ??CH ????6.90 ????H-9，H-12
????12 ????113.572 ??CH ????6.70 ????H-11
????13 ????157.867 ??q ????13-OCH 3， ????H-11，H-15，H-12，H-14
????13-OCH 3 ????55.206 ??CH 3 ????3.73
????14 ????113.572 ??CH ????6.70 ????H-15
????15 ????130.503 ??CH ????6.90 ????H-9，H-14
????1′ ????64.418 ??CH ????3.57 ????H-8′2′-CH 3，H-9′
????2′-CH 3 ????42.748 ??CH 3 ????2.51
????3′ ????46.952 ??CH 2 ????3.16，2.78 ????2′-CH 3
????4′ ????26.115 ??CH 2 ????2.87，2.66 ????H-3′，H-5′
????4′a ????130.699 ??q ????H-1′，H-5′
????5′ ????112.455 ??CH ????6.64 ????H-4′eq
????6′ ????149.090 ??q ????H-5′，H-8′，6′-OCH 3
????6′-OCH 3 ????55.821 ??CH 3 ????3.79
????7′ ????142.866 ????q ????H-5′，H-8′
????8′ ????120.067 ????CH ????6.30
????8′a ????130.383 ????q ????H-1′，H-5′
????9′ ????40.679 ????CH 2 ????2.93，2.80 ????H-11′，H-15′
????10′ ????131.810 ????q ????H-14′
????11′ ????119.085 ????CH ????6.46 ????H-15′，H-9′
????12′ ????144.577 ????q ????H-11′，H-14′
????13′ ????145.254 ????q ????H-15′，H-11′
????14′ ????115.427 ????CH ????6.81
????15′ ????125.239 ????CH ????6.72 ????H-11′
aDEPT 135. bQuaternary carbon.
Table 5. Neferine 1H-NMR and 1H- 1H COSY spectral data
The atom sequence number ???? 1H-NMRδ，ppm Multiplicity ?? 1H- 1H?COSY
??1 ????3.62 ????1H，m ????H-9
??2-CH 3 ????2.48 ????3H，s
??3ax ????2.73 ????1H，m ????H-3eq
??3eq ????3.13 ????1H，m ????H-3ax，H-4
??ax ????2.62 ????1H，m ????H-4eq，H-3
??eq ????2.79 ????1H，m ????H-4ax，H-3
??5 ????6.61 ????1H，s
??6-OCH 3 ????3.79 ????3H，s
??8 ????6.34 ????1H，s
??9ax ????2.77 ????1H，m ????H-9eq，H-1
??9eq ????2.98 ????1H，m ????H-9ax，H-1
??11 ????6.89 ????1H，s ????H-12
??12 ????6.67 ????1H，d，J＝7.06 ????H-11
??13-OCH 3 ????3.70 ????3H，s
??14 ????6.67 ????1H，d，J＝7.06 ????H-15
??15 ????6.89 ????1H，d，J＝7.06 ????H-14
??1′ ????3.62 ????1H，m ????H-9′
??2′-CH 3 ????2.45 ????3H，s
??3′ax ????2.73 ????1H，m ????H-3′eq，H-4′
??3′eq ????3.08 ????1H，m ????H-3′ax，H-4′
??4′ax ????2.59 ????1H，m ????H-4′eq，H-3′
??4′eq ????2.78 ????1H，m ????H-4′ax，H-3′
??5′ ????6.49 ????1H，s
??6′-OCH 3 ????3.76 ????3H，s
??7′-OCH 3 ????3.52 ????3H，s
??8′ ????5.97 ????1H，s
??9′ax ????2.67 ????1H，m ????H-9′eq，H-1′
??9′eq ????3.06 ????1H，m ????H-9′ax，H-1′
??11′ ????6.51 ????1H，s
??14′ ????6.82 ????1H，d，J＝6.68 ????H-15′
??15′ ????6.66 ????1H，d，J＝6.68 ????H-14′
Table 6. Neferine 13C-NMR and 2D NMR spectral data
The atom sequence number ??? 13C-NMR ??DEPT a ????HMQC ????HMBC
????1 ????64.48 ????CH ????3.62 ????2-CH 3，H-8
????2-CH 3 ????42.766 ????CH 3 ????2.48
????3 ????47.275 ????CH 2 ????3.13，2.73 ????2-CH 3
????4 ????26.194 ????CH 2 ????2.79，2.62 ????H-3，H-5
????4a ????130.466 ????q b ????H-5
????5 ????112.421 ????CH ????6.61
????6 ????149.064 ????q ????H-5，H-6，H-8
????6-OCH 3 ????55.794 ????CH 3 ????3.79
????7 ????142.893 ????q ????H-5，H-8
????8 ????120.094 ????CH ????6.34
????8a ????130.979 ????q ????H-8，H-5，H-4
????9 ????39.94 ????CH 2 ????2.98，2.77 ????H-15，H-11
????10 ????131.506 ????q ????H-12，H14，H-9
????11 ????130.555 ????CH ????6.89 ????H-12，H-9，H-15
????12 ????113.515 ????CH ????6.67 ????H-11，H-14
????13 ????157.877 ????q ????H-11，H-15，H-12， ????H-14，13-OCH 3
????13-OCH 3 ????55.191 ????CH 3 ????3.70
????14 ????113.515 ????CH ????6.67 ????H-12，H-15
????15 ????130.555 ????CH ????6.89 ????H-14，H-9，H-11
????1′ ????64.834 ????CH ????3.62 ????2′-CH 3，H-8′
????2′-CH 3 ????42.521 ????CH 3 ????2.45
????3′ ????46.644 ????CH 2 ????3.08，2.73 ????2′-CH 3
????4′ ????25.214 ????CH 2 ????2.78，2.59 ????H-5′
????4′a ????129.07 ????q ????H-5′，H-8′
????5′ ????111.211 ????CH ????6.49
????6′ ????147.298 ????q ????H-5′，H-8′，H6′-OCH 3
????6′-OCH 3 ????55.919 ????CH 3 ????3.76
????7′ ????146.367 ????q ????H-5′，7′-OCH 3，H-8′
????7′-OCH 3 ????55.537 ????CH 3 ????3.52
????8′ ????111.066 ????CH ????5.97
????8′a ????125.653 ????q ????H-8′
????9′ ????40.706 ????CH 2 ????3.06，2.67 ????H-15′，H-11′
????10′ ????131.720 ????q ????H-11′，H-15′，H-9′
????11′ ????119.265 ????CH ????6.51 ????H-9′，H-15′
????12′ ????144.995 ????q ????H-14′
????13′ ????145.715 ????q ????H-15′，H-11′
????14′ ????115.879 ????CH ????6.82
????15′ ????125.299 ????CH ????6.66 ????H-9′，H-11′
aDEPT 135. bQuaternary carbon
Embodiment 2:
Plumula Nelumbinis alkaloid crude product preparation method separates the Plumula Nelumbinis alkaloid extract with embodiment 1 with counter current chromatograph.With ethyl acetate, tetracol phenixin, first alcohol and water, be 1: 6: 4 by volume: 1 preparation two-phase solvent, standing demix in liquid separatnig container is told phase up and down.Plumula Nelumbinis alkaloid crude product 5850mg is dissolved in that 15ml goes up mutually and under the 15ml mutually in the formation sample solution.Adverse current chromatogram column internal diameter 4mm, column volume are 1600ml.The phase that below fixes mutually be a mobile phase down mutually, treat that the counter current chromatograph separator column is filled with stationary phase in interior after, sample solution is injected the injection annulus of counter current chromatograph, open counter current chromatograph, rotating speed is 450 to change, flow velocity with 5ml/min injects mobile phase, and the UV-detector monitoring flow point by online detects wavelength 280nm, collect, merge effluent, obtain liensinine, Isoliensinine and Neferine respectively, its purity is respectively 98.52%, 97.12%, 99.54%, see Fig. 4.
Embodiment 3:
Plumula Nelumbinis alkaloid crude product preparation method separates the Plumula Nelumbinis alkaloid extract with embodiment 1 with counter current chromatograph.With ether, ethyl acetate, tetracol phenixin, first alcohol and water, be 1: 1: 7.5 by volume: 5: 1 preparation two-phase solvents, standing demix in liquid separatnig container is told phase up and down.Plumula Nelumbinis alkaloid crude product 150mg is dissolved in that 1.5ml goes up mutually and under the 1.5ml mutually in the formation sample solution.Adverse current chromatogram column internal diameter 2mm, column volume are 380ml.The phase that below fixes mutually be a mobile phase down mutually, treat that the counter current chromatograph separator column is filled with stationary phase in interior after, sample solution is injected the injection annulus of counter current chromatograph, open counter current chromatograph, rotating speed is 550 to change, flow velocity with 1.5ml/min injects mobile phase, UV-detector monitoring flow point by online detects wavelength 280nm, collects, merges effluent, obtain liensinine, Isoliensinine and Neferine respectively, its purity is respectively 96.54%, 95.13%, and 97.27%.
Embodiment 4:
Plumula Nelumbinis alkaloid crude product preparation method separates the Plumula Nelumbinis alkaloid extract with embodiment 1 with counter current chromatograph.With normal hexane and water, be 10: 7 preparation two-phase solvents by volume, standing demix in liquid separatnig container is told phase up and down.Plumula Nelumbinis alkaloid crude product 55mg is dissolved in that 1ml goes up mutually and under the 1ml mutually in the formation sample solution.Adverse current chromatogram column internal diameter 1.6mm, column volume are 280ml.The phase that below fixes mutually be a mobile phase down mutually, treat that the counter current chromatograph separator column is filled with stationary phase in interior after, sample solution is injected the injection annulus of counter current chromatograph, open counter current chromatograph, rotating speed is 550 to change, flow velocity with 1ml/min injects mobile phase, UV-detector monitoring flow point by online detects wavelength 280nm, collects, merges effluent, obtain liensinine, Isoliensinine and Neferine respectively, its purity is respectively 97.24%, 95.45%, and 96.33%.

Claims (3)

1. from Plumula Nelumbinis, separate the method for preparing liensinine, Isoliensinine and Neferine, it is characterized in that may further comprise the steps:
1) with Plumula Nelumbinis powder with organic solvent lixiviate repeatedly, extracting solution is merged, is condensed into medicinal extract, gained medicinal extract is lower than the dissolving of 20% aqueous acid with concentration, filters, filtrate transfers PH to alkalescence, separates out throw out, obtains the Plumula Nelumbinis alkaloid crude product; Perhaps Plumula Nelumbinis powder directly is lower than 20% aqueous acid lixiviate with concentration, filters, filtrate transfers PH to alkalescence, separates out throw out, obtains the Plumula Nelumbinis alkaloid crude product;
2) will be used for isolating solvent and pour skimmer into, and form immiscible two-phase solvent up and down, the biphase volume ratio is 0.5-2.0 up and down;
3) the Plumula Nelumbinis alkaloid crude product is dissolved in step 2) two-phase solvent in, obtain the Plumula Nelumbinis alkaloid sample solution;
4) with step 2) one in the two-phase solvent inject in the counter current chromatograph separator column mutually as stationary phase, the Plumula Nelumbinis alkaloid crude product solution of step 3) gained is injected the injection annulus of counter current chromatograph, open counter current chromatograph, another of injection two-phase solvent made mobile phase mutually, Plumula Nelumbinis alkaloid just carries out successive and distributes in counter current chromatograph, by detection at thread detector, collect, merge effluent, obtain liensinine, Isoliensinine and the Neferine of purifying respectively.
2. the method for preparing liensinine, Isoliensinine and Neferine of separating from Plumula Nelumbinis according to claim 1 is characterized in that the said organic solvent of step 1) is normal paraffin, halohydrocarbon, Fatty Alcohol(C12-C14 and C12-C18), aliphatic ketone, fatty ester or ether.
3. the method prepare liensinine, Isoliensinine and Neferine of separating from Plumula Nelumbinis according to claim 1 is characterized in that step 2) saidly be used for two or more the mixture that isolating solvent is normal paraffin, halohydrocarbon, Fatty Alcohol(C12-C14 and C12-C18), aliphatic ketone, fatty ester, ethers or water.
CN 200410025198 2004-06-09 2004-06-09 Preparation of liensinine, isoliensinine and methylliensinine extracted from lotus seed Expired - Fee Related CN1286816C (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101786984A (en) * 2010-03-31 2010-07-28 聊城大学 Method for extracting liensinine, isoliensinine and neferine from lotus plumule
CN102657652A (en) * 2011-03-10 2012-09-12 李宏 Novel uses of bisbenzylisoquinoline alkaloid derivative or analogue of general formula I
CN101862331B (en) * 2009-04-14 2013-05-22 中国中医科学院中药研究所 New use of methyl liensinine
CN104744367A (en) * 2015-04-03 2015-07-01 辽宁大学 Flash extraction method of lotus plumule alkaloid
CN114276293A (en) * 2022-01-09 2022-04-05 福建中医药大学 Preparation and purification method of neferine perchlorate

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101862331B (en) * 2009-04-14 2013-05-22 中国中医科学院中药研究所 New use of methyl liensinine
CN101786984A (en) * 2010-03-31 2010-07-28 聊城大学 Method for extracting liensinine, isoliensinine and neferine from lotus plumule
CN101786984B (en) * 2010-03-31 2011-09-07 聊城大学 Method for extracting liensinine, isoliensinine and neferine from lotus plumule
CN102657652A (en) * 2011-03-10 2012-09-12 李宏 Novel uses of bisbenzylisoquinoline alkaloid derivative or analogue of general formula I
CN104744367A (en) * 2015-04-03 2015-07-01 辽宁大学 Flash extraction method of lotus plumule alkaloid
CN114276293A (en) * 2022-01-09 2022-04-05 福建中医药大学 Preparation and purification method of neferine perchlorate
CN114276293B (en) * 2022-01-09 2024-01-26 福建中医药大学 Preparation and purification method of methyl lotus plumule perchlorate

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