CN102086236A - Method for extracting and separating in vivo alkali-soluble polysaccharide from old water shield leaves - Google Patents

Abstract

The invention discloses a method for extracting and separating in vivo alkali-soluble polysaccharide from old water shield leaves, mainly a technology for extracting and separating in vivo polysaccharide from the old leaves remained after processing tender water shield leaves and from the old water shield leaves which are not used in the filed in later October. The method comprises the following technical steps: drying and smashing fresh old water shield leaves; extracting smashed leaves with hot water, extracting the residue after water extraction with diluted alkaline solution, centrifuging and collecting supernate; concentrating the supernate at reduced pressure, carrying out ethanol precipitation, centrifuging and collecting precipitate; and carrying out purifying and spray drying on the precipitate, so as to obtain the alkali-soluble polysaccharide in the old water shield leaves. The method is low in cost, high in product yield, environment-friendly and suitable for industrial production.

Description

A kind of from water shield Lao Ye the method for caustic solubility polysaccharide in the extraction separation body

Technical field

The invention belongs to the natural product extracting technique field, be specifically related to a kind of from water shield Lao Ye the method for caustic solubility polysaccharide in the extraction separation body.

Background technology

Water shield, a kind of original perianth subclass Nymphaeceae, water shield belongs to perennial fresh water aquatic herbaceous plant [Braseniaschreberi].Have another name called Herba Calthae Membranaceae, lake dish, water certain herbaceous plants with big flowers, reveal certain herbaceous plants with big flowers.This plant mainly is distributed in Zhejiang, Jiangsu two province's Taihu Lake basin, and western Lichuan City, Hubei Province is domestic.Late April to late October plucks and has the slightly tender of leaf roll.Traditional applicating history and modern pharmacological research to water shield among the people shows that the water shield polysaccharide has effects such as decreasing cholesterol, hypoglycemic, reducing blood-fat, anti-oxidant, anti-inflammatory, antiviral and immunomodulatory usually, polysaccharide has certain reducing power in the water shield tender leaf body, simultaneously ultra-oxygen anion free radical and hydroxy radical qiao is also had certain removing ability.At present about the extraction of water shield polysaccharide, extraction, the separation that separation method research mainly concentrates on the external polysaccharide of water shield tender leaf.Relevant patent documentation has: the gel technique of water shield (CN:03113443.2), and water shield polysaccharide viscose glue is produced bacterium and water shield polysaccharide viscose glue production technique (CN:94112514.9); The pertinent literature of water shield polysaccharide aspect: the 1. extraction of water shield polysaccharide, separation and some bioactive research (Wang Shuru etc., China Medicine University's journal, 1987, No.3); 2. the extraction of water shield polysaccharide and Study on Functional Properties thereof (Wang Qian etc., Southern Yangtze University, 2009); The 3. initial analysis of external colloid separation of water shield and moiety (Zhou Yifeng, etc., food and fermentation industries, 2005, No.4); The 4. IR spectroscopic analysis in the research of water shield polysaccharide extracting process (Gu Xiaoman etc., Wuhan Institute of Chemical Technology's journal, 1997, No.1); 5. water shield water-soluble polysaccharide extraction process optimization research (Tang Qiaoyu etc., northern gardening, 2008, No.3); 6. the IR in the water shield polysaccharide extracting process, UV spectroscopic analysis (Yu Shixin etc., the Hubei chemical industry, 1996, No.3); 7. the extraction and separation process technology of water shield polysaccharide (today science and technology, 1999, NO.1); 8. the glucuronic acid in the spectrophotometry water shield polysaccharide (Yu Shixin etc., analytical chemistry, 1997, No.9).

With the water shield tender leaf is that raw material carries out the external polysaccharide extraction of water shield, separates, and its resource utilization is low, and added value is not high, certainly will influence the economic benefit of comprehensive development and utilization water shield resource.

Summary of the invention

The object of the present invention is to provide a kind of from remaining Lao Ye in water shield processing back and the late October water shield Lao Ye that no longer utilizes of field the method for polysaccharide in the extraction separation body.

The present invention a kind of from water shield Lao Ye the method for caustic solubility polysaccharide in the extraction separation body, it is characterized in that may further comprise the steps:

A. raw material is handled: fresh water shield Lao Ye drying, pulverized 100 mesh sieves;

B. lixiviate: the water shield raw material powder is with 1: 20 solid-liquid ratio of weight, through 100 ℃ of hot water extraction 60min, centrifugal 15min under 4000rpm repeats to extract 1 time to filter residue while hot, filter residue is with 1: 20 solid-liquid ratio of weight, through 0.02M NaOH solution, 80 ℃ of temperature are extracted 60min, the centrifugal 15min of 4000rpm, collect supernatant liquor, filter residue is repeated to extract 1 time, merge supernatant liquor twice;

C. concentrating and separating: supernatant liquor is evaporated to the 1/3-1/5 of original volume under 50-60 ℃, re-adjustment pH is about 7, adds 95% ethanol of 3 times of volumes then, and the limit adds the ethanol limit stirs, place 8-24h after, the centrifugal 15min of 4000rpm, collecting precipitation;

D. washing: precipitation gets water shield caustic solubility Crude polysaccharides after 95% washing with alcohol;

E. purifying: the 0.02M NaOH dilute alkaline soln that Crude polysaccharides adds 20 times of weight redissolves; With sevag method deproteinated; Re-adjustment polysaccharide soln pH to pH8 after 50 ℃ isopyknic 20% hydrogen peroxide of dropping decolours down, adds 95% ethanol sedimentation polysaccharide of 3 times of volumes, and is centrifugal, collecting precipitation; The 0.02M NaOH solution that adds 20 times of weight again redissolves and precipitates, get polysaccharide soln, 95% ethanol sedimentation polysaccharide with 3 times of volumes of polysaccharide soln, centrifugal, collecting precipitation, with the 0.02M NaOH solution dissolution precipitation of 20 times of weight, regulating pH is about 7.0, spraying drying while stirring then, water shield caustic solubility polysaccharide.

The present invention is directed to Lao Ye remaining and the late October water shield Lao Ye that no longer utilizes of field from water shield processing back, after adopting the hot water lixiviate, extract caustic solubility polysaccharide in the water shield Lao Ye body with dilute alkaline soln again, through repeatedly alcohol precipitation, deproteinated, decolouring, spraying drying, obtain water shield caustic solubility polysaccharide, this method cost is low, the extraction yield is higher, environmental protection is fit to the industrialization continuous production, can realize the comprehensive development and utilization of water shield resource and the raising of economic benefit.

Embodiment

The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited thereto.

Embodiment 1

Raw material is handled: fresh water shield Lao Ye drying, pulverized 100 mesh sieves, and standby;

The lixiviate of caustic solubility polysaccharide: raw material powder 0.5kg, add 10L distilled water, be heated to 100 ℃, extract 60min, the centrifugal 15min of 4000rpm while hot is to filter residue adding distil water 10L, be heated to and repeat to extract under 100 ℃ 1 time, filter residue adds 10L 0.02M NaOH solution, is heated to 80 ℃, extracts 60min, the centrifugal 15min of 4000rpm, collect supernatant liquor, filter residue is repeated to extract 1 time, merge supernatant liquor twice;

Concentrating and separating: supernatant liquor is evaporated to 1/3 of original volume under 60 ℃, re-adjustment pH is 7, adds 95% ethanol of 3 times of volumes then, and the limit adds the ethanol limit stirs, behind the placement 24h, and the centrifugal 15min of 4000rpm, collecting precipitation;

Purifying: precipitation is after 95% washing with alcohol, get water shield caustic solubility Crude polysaccharides, add 1000ml 0.02M NaOH solution redissolve polysaccharide liquid, with sevag method deproteinated, promptly with the liquid glucose of 3: 1 (v/v): mixed solution (4 chloroforms: 1 propyl carbinol, v/v) deproteinated is 1 time, re-adjustment polysaccharide soln pH to pH8 after 50 ℃ isopyknic 20% hydrogen peroxide of dropping decolours down, adds 95% ethanol sedimentation polysaccharide of 3 times of volumes, centrifugal, collecting precipitation; After adding the redissolution of 1000ml 0.02M NaOH solution again, the 95% ethanol sedimentation polysaccharide that adds 3 times of volumes, centrifugal, collecting precipitation adds the dissolving of 1000ml 0.02M NaOH solution, and regulating pH is about 7.0, spraying drying while stirring then, get the water shield caustic solubility polysaccharide of preliminary purification, about 11% (glucuronic acid is made standard specimen, and the carbazole method is measured) of product yield.

Embodiment 2

Raw material is handled: fresh water shield Lao Ye drying, pulverized 100 mesh sieves, and standby;

The lixiviate of caustic solubility polysaccharide: raw material powder 1kg, add 10L distilled water, be heated to 100 ℃, extract 60min, the centrifugal 15min of 4000rpm while hot is to filter residue adding distil water 10L, be heated to and repeat to extract under 100 ℃ 1 time, add 20L 0.02M NaOH solution in the filter residue, be heated to 80 ℃, extract 60min, the centrifugal 15min of 4000rpm, collect supernatant liquor, filter residue is repeated to extract 1 time, merge supernatant liquor twice;

Concentrating and separating: supernatant liquor is evaporated to 1/3 of original volume under 60 ℃, re-adjustment pH is 7, adds 95% ethanol of 3 times of volumes then, and the limit adds the ethanol limit stirs, behind the placement 24h, and the centrifugal 15min of 4000rpm, collecting precipitation;

Purifying: precipitation is after 95% washing with alcohol, get water shield caustic solubility Crude polysaccharides, add 2000ml 0.02M NaOH solution redissolve polysaccharide liquid, with sevag method deproteinated, promptly with the liquid glucose of 3: 1 (v/v): mixed solution (4 chloroforms: 1 propyl carbinol, v/v) deproteinated is 1 time, re-adjustment polysaccharide soln pH to pH8 after 50 ℃ isopyknic 20% hydrogen peroxide of dropping decolours down, adds 95% ethanol sedimentation polysaccharide of 3 times of volumes, centrifugal, collecting precipitation; After adding the redissolution of 2000ml 0.02M NaOH solution again, the 95% ethanol sedimentation polysaccharide that adds 3 times of volumes, centrifugal, collecting precipitation adds the dissolving of 2000ml 0.02M NaOH solution, and regulating pH is about 7.0, spraying drying while stirring then, get the water shield caustic solubility polysaccharide of preliminary purification, about 11% (glucuronic acid is made standard specimen, and the carbazole method is measured) of product yield.

Embodiment 3

Raw material is handled: fresh water shield Lao Ye drying, pulverized 100 mesh sieves, and standby;

The lixiviate of caustic solubility polysaccharide: raw material powder 2kg, add 10L distilled water, be heated to 100 ℃, extract 60min, the centrifugal 15min of 4000rpm while hot is to filter residue adding distil water 10L, be heated to and repeat to extract under 100 ℃ 1 time, add 40L 0.02M NaOH solution in the water shield residue of filter residue after water is carried, be heated to 80 ℃, extract 60min, the centrifugal 15min of 4000rpm, collect supernatant liquor, filter residue is repeated to extract 1 time, merge supernatant liquor twice;

Concentrating and separating: supernatant liquor is evaporated to 1/3 of original volume under 60 ℃, re-adjustment pH is 7, adds 95% ethanol of 3 times of volumes then, and the limit adds the ethanol limit stirs, behind the placement 24h, and the centrifugal 15min of 4000rpm, collecting precipitation;

Purifying: precipitation is after 95% washing with alcohol, get water shield caustic solubility Crude polysaccharides, add 4000ml 0.02M NaOH solution redissolve polysaccharide liquid, with sevag method deproteinated, promptly with the liquid glucose of 3: 1 (v/v): mixed solution (4 chloroforms: 1 propyl carbinol, v/v) deproteinated is 1 time, re-adjustment polysaccharide soln pH to pH8 after 50 ℃ isopyknic 20% hydrogen peroxide of dropping decolours down, adds 95% ethanol sedimentation polysaccharide of 3 times of volumes, centrifugal, collecting precipitation; After adding the redissolution of 4000ml 0.02M NaOH solution again, the 95% ethanol sedimentation polysaccharide that adds 3 times of volumes, centrifugal, collecting precipitation adds the dissolving of 4000ml 0.02M NaOH solution, and regulating pH is about 7.0, spraying drying while stirring then, get the water shield caustic solubility polysaccharide of preliminary purification, about 11% (glucuronic acid is made standard specimen, and the carbazole method is measured) of product yield.

Claims (1)

1. method of caustic solubility polysaccharide in the extraction separation body from water shield Lao Ye is characterized in that may further comprise the steps:

A. raw material is handled: fresh water shield Lao Ye drying, pulverized 100 mesh sieves;

B. lixiviate: the water shield raw material powder is with 1: 20 solid-liquid ratio of weight, through 100 ℃ of hot water extraction 60min, centrifugal 15min under 4000rpm repeats to extract 1 time to filter residue while hot, filter residue is with 1: 20 solid-liquid ratio of weight, through 0.02M NaOH solution, 80 ℃ of temperature are extracted 60min, the centrifugal 15min of 4000rpm, collect supernatant liquor, filter residue is repeated to extract 1 time, merge supernatant liquor twice;

C. concentrating and separating: supernatant liquor is evaporated to the 1/3-1/5 of original volume under 50-60 ℃, re-adjustment pH is about 7, adds 95% ethanol of 3 times of volumes then, and the limit adds the ethanol limit stirs, place 8-24h after, the centrifugal 15min of 4000rpm, collecting precipitation;

D. washing: precipitation gets water shield caustic solubility Crude polysaccharides after 95% washing with alcohol;

E. purifying: the 0.02M NaOH dilute alkaline soln that Crude polysaccharides adds 20 times of weight redissolves; With sevag method deproteinated; Re-adjustment polysaccharide soln pH to pH8 after 50 ℃ isopyknic 20% hydrogen peroxide of dropping decolours down, adds 95% ethanol sedimentation polysaccharide of 3 times of volumes, and is centrifugal, collecting precipitation; The 0.02M NaOH solution that adds 20 times of weight again redissolves and precipitates, get polysaccharide soln, 95% ethanol sedimentation polysaccharide with 3 times of volumes of polysaccharide soln, centrifugal, collecting precipitation, with the 0.02M NaOH solution dissolution precipitation of 20 times of weight, regulating pH is about 7.0, spraying drying while stirring then, water shield caustic solubility polysaccharide.