CN104557505A - Separation method of effective components of sharpleaf galangal fruit aqueous extract - Google Patents

Separation method of effective components of sharpleaf galangal fruit aqueous extract Download PDF

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CN104557505A
CN104557505A CN201310504602.6A CN201310504602A CN104557505A CN 104557505 A CN104557505 A CN 104557505A CN 201310504602 A CN201310504602 A CN 201310504602A CN 104557505 A CN104557505 A CN 104557505A
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water
acetonitrile
formic acid
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sharpleaf galangal
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CN104557505B (en
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王宗权
常青鲜
贾继明
乔莉
姜新刚
孟作环
秦拢
马艳玲
宋剑
裴彩云
王贵金
蔡燕
周梦夏
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Hebei Yiling Pharmaceutical Research Institute Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/78—Separation; Purification; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
    • C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins

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  • Oil, Petroleum & Natural Gas (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses a separation method of effective components of a sharpleaf galangal fruit aqueous extract, belonging to the field of pharmaceutical analysis. The separation method comprises the following steps: (A) fetching dried sharpleaf galangal fruit, grinding, carrying out reflux extraction with water, combining filtrate, carrying out reduced pressure distillation to obtain extractum, dissolving the extractum into distilled water, respectively extracting by virtue of petroleum ether, ethyl acetate and normal butanol, and recycling solvents, so as to obtain ethyl acetate extract dry powder; (B) dissolving the dry powder obtain in the step (A), filtering, and processing by virtue of a sephadex LH20 gel column, so as to obtain six components from F1 to F6; and (C) carrying out HPLC separation and purification on the components from F2 to F6, so as to obtain compounds including protocatechualdehyde, protocatechuic acid, p-coumaric acid, (-)-epicatechin and (+)-catechinic acid. According to the separation method, the effective active components are separated and extracted from the sharpleaf galangal fruit, the structures of the effective components are further defined, the basis is provided for later medication, and required components can be extracted according to different requirements.

Description

A kind of separation method of effective constituent of Sharpleaf Galangal Fruit aqueous extract
Technical field
The invention belongs to pharmaceutical analysis field, be specifically related to the separation and purification of Effective Component of Chinese Medicine.
Background technology
Sharpleaf Galangal Fruit is the dry mature fruit of Zingiber (Zingiberaceae) Alpinia (Alpinia Roxb.) plant intelligence development (A.oxyphylla Miq.), and be one of Chinese four great Nan medicines, main product, in the ground such as Hainan and Guangdong, is parts of generic medicinal plants.Sharpleaf Galangal Fruit has warm kidney controlling nocturnal emission with astringent drugs contracting urine, warming spleen and stopping diarrhaly takes the photograph the effects such as saliva, is clinically used for enuresis due to deficiency of the kidney, frequent micturition, and seminal emission gonorrhoea, malaria is had loose bowels, crymodynia in abdomen, the many salivas of mouth.Modern pharmacological research shows: Sharpleaf Galangal Fruit aqueous extract has neuroprotective and antioxygenation, and Sharpleaf Galangal Fruit also has cardiac stimulant, Ca2+ overloading, anticancer and improve the effects such as kidney in addition.Bibliographical information has isolated the Multiple components such as diaryl heptane class, sesquiterpenoids, flavonoid, steroidal and organic acid from Sharpleaf Galangal Fruit.But report is had no to the research of the aqueous extract chemical composition of Sharpleaf Galangal Fruit.
Summary of the invention
The object of the invention is to provide a kind of separation method of effective constituent of Sharpleaf Galangal Fruit aqueous extract, simple to operate.
The technical solution adopted in the present invention is:
A separation method for the effective constituent of Sharpleaf Galangal Fruit aqueous extract, described method is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, use water refluxing extraction, merging filtrate after pulverizing, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, uses sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtains ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with ethanol gradient elution, collects each section of effluent, obtain 6 component F 1~ F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid, (-)-l-Epicatechol, (+)-catechin.
As optimal way, described method is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, by 8-10 times of water gaging refluxing extraction 2 times after pulverizing, each 1.5 hours, merging filtrate, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, use sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtain ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with 20-40% ethanol gradient elution, collect each section of effluent, 20% ethanol elution obtains F 1, 30% ethanol elution obtains F 2, F 3, F 4, 40% ethanol elution obtains F 5, F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, component F 2~ F 4moving phase be acetonitrile-0.3% formic acid water, gradient elution: 0 ~ 10 min, 5% acetonitrile, the 0.3% formic acid water of 95%, 10 ~ 20 min, 5% ~ 12% acetonitrile, the 0.3% formic acid water of 95-88%, 20 ~ 40 min, 12% acetonitrile, the 0.3% formic acid water of 88%, 40 ~ 60 min, 12% ~ 40% acetonitrile, the 0.3% formic acid water of 88-60%, 60 ~ 70 min, 40% ~ 100% acetonitrile, the 0.3% formic acid water of 60-0%, 70 ~ 80 min, 100% acetonitrile, the 0.3% formic acid water of 0%, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid; F 5~ F 6moving phase be acetonitrile-water, gradient elution: 0 ~ 10 min 5% acetonitrile, 95% water, 10 ~ 20 min 5% ~ 12% acetonitriles, 95-88% water, 20 ~ 40 min, 12% acetonitrile, 88% water, 40 ~ 60 min, 12% ~ 40% acetonitrile, 88-60% water, 60 ~ 70 min, 40% ~ 100% acetonitrile, 60-0% water, 70 ~ 80 min, 100% acetonitrile, 0% water, obtains compound (-)-l-Epicatechol, (+)-catechin.
As optimal way, in described step C, chromatographic column is C18, and specification is 10 × 250 mm, 5 μm, and in step C, elution flow rate is 1 mL/min.
Owing to have employed technique scheme, the technical progress that the present invention obtains is:
The present invention's separation and Extraction from Sharpleaf Galangal Fruit goes out effective activeconstituents, specify that the structure of effective constituent further, for later medication provides basis, can extract the composition needed according to no needs.
Embodiment
Below in conjunction with embodiment, the present invention is described in further details:
Nucleus magnetic resonance adopts Varian uNITYiNOVA600 superconduction nuclear magnetic resonance spectrometer measures (in TMS mark); TOF-MS adopts AB Sciex 5600 high resolution magnetic mass spectrometer to measure; Fusing point adopts YRT-3 melting point apparatus to measure; Preparation HPLC adopts LC-8A preparative scale chromatography instrument, SPD-M10A detector, SIL-10A automatic sampler and FRC-10A automatic safety part collector (Japanese Shimadzu Corporation); Half preparative chromatography post is SunFire Prep C18 (10 × 250 mm, 5 μm) (waters company of the U.S.); Sephadex LH-20 dextrane gel (GE company); Acetonitrile is chromatographically pure, and all the other are analytical pure.
Sharpleaf Galangal Fruit is purchased from Anhui prepared slices of Chinese crude drugs company limited of wellspring group, and the place of production is Guangdong, is accredited as through combination of Chinese tradiational and Western medicine Medicine Research Academy resource room, Hebei province a. oxyphyllafruit.
embodiment 1
The separation method of the effective constituent in Sharpleaf Galangal Fruit Aqueous extracts, is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, pulverize rear 8 times of water gaging refluxing extraction 2 times, each 1.5 hours, merging filtrate, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, use sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtain ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with 20-40% ethanol gradient elution, collect each section of effluent, 20% ethanol elution obtains F 1, 30% ethanol elution obtains F 2, F 3, F 4, 40% ethanol elution obtains F 5, F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, and chromatographic column is C18, and specification is 10 × 250 mm, 5 μm, component F 2~ F 4moving phase be acetonitrile-0.3% formic acid water, gradient elution: 0 ~ 10 min, 5% acetonitrile, the 0.3% formic acid water of 95%, 10 ~ 20 min, 5% ~ 12% acetonitrile, the 0.3% formic acid water of 95-88%, 20 ~ 40 min, 12% acetonitrile, the 0.3% formic acid water of 88%, 40 ~ 60 min, 12% ~ 40% acetonitrile, the 0.3% formic acid water of 88-60%, 60 ~ 70 min, 40% ~ 100% acetonitrile, the 0.3% formic acid water of 60-0%, 70 ~ 80 min, 100% acetonitrile, the 0.3% formic acid water of 0%, flow velocity is 1 mL/min, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid; F 5~ F 6moving phase be acetonitrile-water, gradient elution: 0 ~ 10 min 5% acetonitrile, 95% water, 10 ~ 20 min 5% ~ 12% acetonitriles, 95-88% water, 20 ~ 40 min, 12% acetonitrile, 88% water, 40 ~ 60 min, 12% ~ 40% acetonitrile, 88-60% water, 60 ~ 70 min, 40% ~ 100% acetonitrile, 60-0% water, 70 ~ 80 min, 100% acetonitrile, 0% water, flow velocity is 1 mL/min, obtains compound (-)-l-Epicatechol, (+)-catechin.
embodiment 2
The separation method of the effective constituent in Sharpleaf Galangal Fruit Aqueous extracts, is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, pulverize rear 10 times of water gaging refluxing extraction 2 times, each 1.5 hours, merging filtrate, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, use sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtain ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with 20-40% ethanol gradient elution, collect each section of effluent, 20% ethanol elution obtains F 1, 30% ethanol elution obtains F 2, F 3, F 4, 40% ethanol elution obtains F 5, F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, and chromatographic column is C18, and specification is 10 × 250 mm, 5 μm, component F 2~ F 4moving phase be acetonitrile-0.3% formic acid water, gradient elution: 0 ~ 10 min, 5% acetonitrile, the 0.3% formic acid water of 95%, 10 ~ 20 min, 5% ~ 12% acetonitrile, the 0.3% formic acid water of 95-88%, 20 ~ 40 min, 12% acetonitrile, the 0.3% formic acid water of 88%, 40 ~ 60 min, 12% ~ 40% acetonitrile, the 0.3% formic acid water of 88-60%, 60 ~ 70 min, 40% ~ 100% acetonitrile, the 0.3% formic acid water of 60-0%, 70 ~ 80 min, 100% acetonitrile, the 0.3% formic acid water of 0%, flow velocity is 1 mL/min, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid; F 5~ F 6moving phase be acetonitrile-water, gradient elution: 0 ~ 10 min 5% acetonitrile, 95% water, 10 ~ 20 min 5% ~ 12% acetonitriles, 95-88% water, 20 ~ 40 min, 12% acetonitrile, 88% water, 40 ~ 60 min, 12% ~ 40% acetonitrile, 88-60% water, 60 ~ 70 min, 40% ~ 100% acetonitrile, 60-0% water, 70 ~ 80 min, 100% acetonitrile, 0% water, flow velocity is 1 mL/min, obtains compound (-)-l-Epicatechol, (+)-catechin.
embodiment 3
The separation method of the effective constituent in Sharpleaf Galangal Fruit Aqueous extracts, is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, pulverize rear 9 times of water gaging refluxing extraction 2 times, each 1.5 hours, merging filtrate, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, use sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtain ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with 20-40% ethanol gradient elution, collect each section of effluent, 20% ethanol elution obtains F 1, 30% ethanol elution obtains F 2, F 3, F 4, 40% ethanol elution obtains F 5, F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, and chromatographic column is C18, and specification is 10 × 250 mm, 5 μm, component F 2~ F 4moving phase be acetonitrile-0.3% formic acid water, gradient elution: 0 ~ 10 min, 5% acetonitrile, the 0.3% formic acid water of 95%, 10 ~ 20 min, 5% ~ 12% acetonitrile, the 0.3% formic acid water of 95-88%, 20 ~ 40 min, 12% acetonitrile, the 0.3% formic acid water of 88%, 40 ~ 60 min, 12% ~ 40% acetonitrile, the 0.3% formic acid water of 88-60%, 60 ~ 70 min, 40% ~ 100% acetonitrile, the 0.3% formic acid water of 60-0%, 70 ~ 80 min, 100% acetonitrile, the 0.3% formic acid water of 0%, flow velocity is 1 mL/min, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid; F 5~ F 6moving phase be acetonitrile-water, gradient elution: 0 ~ 10 min 5% acetonitrile, 95% water, 10 ~ 20 min 5% ~ 12% acetonitriles, 95-88% water, 20 ~ 40 min, 12% acetonitrile, 88% water, 40 ~ 60 min, 12% ~ 40% acetonitrile, 88-60% water, 60 ~ 70 min, 40% ~ 100% acetonitrile, 60-0% water, 70 ~ 80 min, 100% acetonitrile, 0% water, flow velocity is 1 mL/min, obtains compound (-)-l-Epicatechol, (+)-catechin.
Structural Identification
Compound 1: brown powder, C 7h 6o 3, mp.154 ~ 155 DEG C.UV (CH 3OH) λ max(nm) 231.0、279.5、310.5。TOF-MS m/z：139.0394[M+H] +。 1H-NMR（DMSO- d 6，600 MH Z）δ：6.91 (1H，brs，H-5) ，7.23 (1H，s，H-2)，7.26 (1H，d， J=6.6 H Z，H-6)，9.69 (1H，s，CHO)。 13C-NMR（DMSO- d 6，150 MH Z） δ：128.7 (C-1)，114.3 (C-2)，145.9 (C-3)，152.3 (C-4)，115.5 (C-5)，124.5 (C-6)，191.0 (CHO)。Above spectral data and bibliographical information basically identical.Therefore authenticating compound 1 is rancinamycin IV (protocatechuic aldehyde).
Compound 2: white, needle-shaped crystals (methyl alcohol), C 7h 6o 4, mp.203 ~ 205 DEG C.UV (CH 3OH) λ max(nm) 231.0、279.5、310.5。TOF-MS m/z：155.0337[M+H] +。 1H-NMR（DMSO- d 6，600 MH Z）δ：6.79 (1H，brs，H-5) ，7.28 (1H，d， J=7.2 H Z，H-2)，7.34 (1H，brs，H-6)。 13C-NMR（DMSO- d 6，150 MH Z） δ：121.7 (C-1)，116.6 (C-2)，144.9 (C-3)，150.0 (C-4)，115.1 (C-5)，121.9 (C-6)，167.4 (COOH)。Above spectral data and bibliographical information basically identical.Therefore authenticating compound 2 is Protocatechuic Acid (protocatechuic acid).
Compound 3: brown ceramic powder, C 9h 8o 3, mp.208 ~ 210 DEG C.UV (CH 3OH) λ max(nm) 309.3。TOF-MS m/z：163.0384[M-H] +。 1H-NMR（DMSO- d 6，600 MH Z）δ：6.28 (1H，d， J=16.2 H Z，H-8) ，6.78 (2H，d， J=8.4 H Z，H-3, H-5)，7.47 (1H，d， J=16.2 H Z，H-7) ，7.62 (2H，d， J=8.4 H Z，H-2, H-6)。 13C-NMR（DMSO- d 6，150 MH Z） δ：125.3 (C-1)，130.0 (C-2, 6)，115.7 (C-3, 5)，159.5(C-4)，143.6 (C-7)，115.7 (C-8)，168.3 (COOH)。Above spectral data and bibliographical information basically identical.Therefore authenticating compound 3 be P-coumaric acid ( p-coumaric acid).
Compound 4: brown ceramic powder, C 15h 14o 6, mp.170 ~ 172 DEG C.UV (CH 3OH) λ max(nm) 278.3。TOF-MS m/z：291.0865[M+H] +。 1H-NMR（DMSO- d 6，600 MH Z）δ：2.47 (1H，dd， J=3.0, 16.2 H Z，H-4β) ，2.67 (1H，dd， J=4.2, 16.2 H Z，H-4α) ，4.00 (1H，brs，H-3) ，4.73 (1H，brs，H-2) ， 5.71 (1H，d， J=2.4 H Z，H-6)，5.89 (1H，d， J=2.4 H Z，H-8)，6.65 (2H，m，H-5'，H-6')，6.88 (1H，d， J=1.8 H Z，H-2')。 13C-NMR（DMSO- d 6，150 MH Z）δ： 78.5 (C-2)，65.4 (C-3)，28.7 (C-4)，156.2 (C-5)，94.5 (C-6)，156.7 (C-7)，95.5 (C-8)，157.0 (C-9)，98.9 (C-10)，131.1 (C-1')，115.4 (C-2')，145.0 (C-3')，144.9 (C-4')，115.2 (C-5')，118.4 (C-6')。Above spectral data and bibliographical information basically identical.Therefore authenticating compound 4 is (-)-l-Epicatechol ((-)-epicatechin).
Compound 5: brown ceramic powder, C 15h 14o 6.UV (CH 3OH) λ max(nm) 278.3。TOF-MS m/z：291.0865[M+H] +。 1H-NMR（DMSO- d 6，600 MH Z）δ：2.34 (1H，dd， J=7.8, 16.2 H Z，H-4β) ，2.65 (1H，dd， J=5.4, 16.2 H Z，H-4α) ，3.81 (1H，m，H-3) ，4.47 (1H， J=7.8 H Z，H-2) ， 5.68 (1H，d， J=2.4 H Z，H-6)，5.88 (1H，d， J=2.4 H Z，H-8)，6.59 (1H，dd， J=1.8, 8.4 H Z，H-6')，6.68 (1H，d， J=7.8 H Z，H-5') ，6.72 (1H，d， J=1.8 H Z，H-2')。 13C-NMR（DMSO- d 6，150 MH Z）δ： 81.0 (C-2)，66.3 (C-3)，27.8 (C-4)，155.3 (C-5)，93.8 (C-6)，156.4 (C-7)，95.0 (C-8)，156.1 (C-9)，99.0 (C-10)，130.5 (C-1')，115.0 (C-2')，144.8 (C-3')，144.8 (C-4')，118.1 (C-5')，114.5 (C-6')。Above spectral data and bibliographical information basically identical.Therefore authenticating compound 5 is (+)-catechin ((+)-catechin).

Claims (3)

1. a separation method for the effective constituent of Sharpleaf Galangal Fruit aqueous extract, is characterized in that described method is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, use water refluxing extraction, merging filtrate after pulverizing, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, uses sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtains ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with ethanol gradient elution, collects each section of effluent, obtain 6 component F 1~ F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid, (-)-l-Epicatechol, (+)-catechin.
2. separation method according to claim 1, is characterized in that described method is made up of following steps:
A, extraction: get dry Sharpleaf Galangal Fruit, by 8-10 times of water gaging refluxing extraction 2 times after pulverizing, each 1.5 hours, merging filtrate, underpressure distillation obtains medicinal extract, is dissolved in by medicinal extract in distilled water, use sherwood oil, ethyl acetate, n-butanol extraction respectively, recycling design, obtain ethyl acetate extract dry powder;
B, gel column are separated: by steps A gained dry powder water dissolution, and filter, upper dextrane gel sephadex LH-20 gel column, with 20-40% ethanol gradient elution, collect each section of effluent, 20% ethanol elution obtains F 1, 30% ethanol elution obtains F 2, F 3, F 4, 40% ethanol elution obtains F 5, F 6;
C, HPLC purifying: by step B gained F 2~ F 6component carries out HPLC separation and purification, component F 2~ F 4moving phase be acetonitrile-0.3% formic acid water, gradient elution: 0 ~ 10 min, 5% acetonitrile, the 0.3% formic acid water of 95%, 10 ~ 20 min, 5% ~ 12% acetonitrile, the 0.3% formic acid water of 95-88%, 20 ~ 40 min, 12% acetonitrile, the 0.3% formic acid water of 88%, 40 ~ 60 min, 12% ~ 40% acetonitrile, the 0.3% formic acid water of 88-60%, 60 ~ 70 min, 40% ~ 100% acetonitrile, the 0.3% formic acid water of 60-0%, 70 ~ 80 min, 100% acetonitrile, the 0.3% formic acid water of 0%, obtains compound rancinamycin IV, Protocatechuic Acid, P-coumaric acid; F 5~ F 6moving phase be acetonitrile-water, gradient elution: 0 ~ 10 min 5% acetonitrile, 95% water, 10 ~ 20 min 5% ~ 12% acetonitriles, 95-88% water, 20 ~ 40 min, 12% acetonitrile, 88% water, 40 ~ 60 min, 12% ~ 40% acetonitrile, 88-60% water, 60 ~ 70 min, 40% ~ 100% acetonitrile, 60-0% water, 70 ~ 80 min, 100% acetonitrile, 0% water, obtains compound (-)-l-Epicatechol, (+)-catechin.
3. the separation method according to any one of claim 1-2, it is characterized in that in described step C, chromatographic column is C18, specification is 10 × 250 mm, 5 μm, and in step C, elution flow rate is 1 mL/min-.
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CN105213920A (en) * 2015-10-10 2016-01-06 河南大学 The application of Fructus Alpiniae Oxyphyllae leaf extract on preparation prevention, treatment hepatic injury health product or medicine
CN108514078A (en) * 2018-03-16 2018-09-11 广东日可威富硒食品有限公司 A kind of kelp vermicelli and preparation method thereof
CN109856254A (en) * 2018-01-11 2019-06-07 广西中医药大学 The method for building up and its finger-print of longan leaf ethyl acetate extract HPLC finger-print
CN110585385A (en) * 2019-10-17 2019-12-20 郑州大学第一附属医院 Extraction method and qualitative analysis method of alpinia oxyphylla sesquiterpenoids

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陈萍等: "益智仁的化学成分及药理活性研究进展", 《现代药物与临床》 *

Cited By (6)

* Cited by examiner, † Cited by third party
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CN105213920A (en) * 2015-10-10 2016-01-06 河南大学 The application of Fructus Alpiniae Oxyphyllae leaf extract on preparation prevention, treatment hepatic injury health product or medicine
CN105213920B (en) * 2015-10-10 2019-02-22 河南大学 Intelligence development leaf extract prevents, treats the application on liver injury medicament in preparation
CN109856254A (en) * 2018-01-11 2019-06-07 广西中医药大学 The method for building up and its finger-print of longan leaf ethyl acetate extract HPLC finger-print
CN108514078A (en) * 2018-03-16 2018-09-11 广东日可威富硒食品有限公司 A kind of kelp vermicelli and preparation method thereof
CN110585385A (en) * 2019-10-17 2019-12-20 郑州大学第一附属医院 Extraction method and qualitative analysis method of alpinia oxyphylla sesquiterpenoids
CN110585385B (en) * 2019-10-17 2021-08-17 郑州大学第一附属医院 Extraction method and qualitative analysis method of alpinia oxyphylla sesquiterpenoids

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