CN1060782C - Preparation and application of sargassum polysaccharide complex - Google Patents

Abstract

The present invention belongs to the technical field of the preparation of biomacromolecule metal complexes, particularly to a calcium polysaccharide complex prepared by the complex reaction of algae and calcium. In order to fully use cheap algae as raw materials and save energy sources, the present invention provides a seaweed calcium polysaccharide complex produced by that dry algae are soaked in alkali water to extract seaweed polysaccharide and the seaweed polysaccharide is in complex reaction with calcium ions under the conditions of normal temperature and normal pressure. The calcium polysaccharide complex is capable of being used for food additives and medicines, used for treating cardiovascular diseases and used as a medicine for treating osteoporosis, and has the advantages of no toxic or side effect, low price, simple preparation method, easy industrialized production and great practical value.

Description

Sargassum polysaccharides calcium complex and preparation method and purposes

The invention belongs to the preparing technical field of biomacromolecule metal complex, particularly relate to the technical field that algae and calcium ion complex reaction prepare the polysaccharide calcium complex.

The biomacromolecule metal complex that biomacromolecule and micro-metal ion react formation under certain condition demonstrates special biological activity, as amino acid, nucleic acid metal complex.The amino acids metal complex compound is made the existing patent of fodder additives.As Methionin and copper complex people such as () the Polish Gdansk Kaczmarek of university, the complex compound that glycine and ferric ion form people such as () the Devore EC of department of chemistry of U.S. Univ Wyoming.Germany's publication number be the complex compound that forms of 2,209,616 medicinal halfcystine and zinc as the zinc supplementation agent, this method is that solder(ing)acid is joined in the halfcystine solution, with NaOH solution adjusting PH and form complex compound.

And for example U.S. Patent number is 3,82l, a kind of glucose of using is disclosed in 192 the specification sheets, maltose, low molecular dextran and ferric chloride reaction form sugared iron complex, and its preparation method mixes with dextran solution with the aqueous solution of ferric ion, and the adding alkaline solution precipitates fully up to iron trichloride, this moment, PH was lower than 7, through stir adding till alkali makes PH rise to 11 above solution to be the homogeneous state, temperature remains on 80 ℃ in the reaction process, not above 95 ℃.For fully being precipitated out also, sugared iron complex to add alcohol solution, through centrifugation, drying, be ground into required particle.The product of describing from above-mentioned patent documentation is with amino acid, glucose, maltose warp and metal ion reacting by heating, whole process all need keep higher temperature of reaction, and the complex compound effect of preparing is single, as prepares sugared iron complex, only is used for the treatment of the piglet hypoferric anemia.

The objective of the invention is to overcome the shortcoming and defect of prior art, in order to make full use of cheap algae resource, and without under the heating condition, marine alga and calcium complex reaction generate the moving foodstuff additive and medical of Sargassum polysaccharides calcium complex conduct with difference in functionality, thereby providing a kind of makes raw material with algae at normal temperatures and pressures and soaks (alkalization) extract polysaccharides in alkali lye, generate the Sargassum polysaccharides calcium complex with the calcium ion complex reaction again, prepare the method for medicinal Sargassum polysaccharides calcium complex with ethanol sedimentation, separation, drying.

Task of the present invention is achieved in that

Adopt natural marine alga as: brown alga, red algae, blue algae, green alga, chrysophyceae etc. make raw material.Put into 0.5-5wt% sodium carbonate solution, 1000-10000 ml for the dry seaweed of 100g in proportion and soaked 12-24 hours, filter and extract polysaccharides liquid, filter residue, again the CaCl of 2 mol/L concentration ₂Solion drips in Sargassum polysaccharides liquid, stirs while dripping, and adds 80-150 ml CaCl altogether ₂Solution, till saturated, to drip the sodium hydroxide solution of 2 mol/L simultaneously, carry out complex reaction, make solution reach at PH=9-11 o'clock and stop to add sodium hydroxide solution, for complex reaction even, continue to stir 0.5-2 hours, generate Sargassum polysaccharides calcium complex solution, the ethanol or other alcohol that add 2 times of liquor capacities more inward, till its product precipitation, and adopt centrifugal or filter method leaches it come, the throw out drying can also adopt mechanical means to be ground into required particle and make the Sargassum polysaccharides calcium complex.Used chemical feedstocks purity all adopts more than the technical purity.

The structure of Sargassum polysaccharides calcium complex is as follows:

M is Ca in the formula ²⁺

Sargassum polysaccharides calcium complex of the present invention is to utilize cheap algae to make raw material, under the condition of pressurization of need not heating, obtains this product, save energy by complex reaction.This product is as foodstuff additive or pharmaceutical cheap, especially as prevention and treatment osteoporosis agents practical value arranged, because adopt natural matter production, can no side effects, and its production method is simple in addition, is easy to scale operation.

Sargassum polysaccharides calcium complex of the present invention can be cooked foodstuff additive; also can be medicinal; it is the medicine of a kind of prevention and treatment osteoporosis; can also prevent and treat cardiovascular disorder; and can protect cerebral neural medicine, its experimental result of effect of Sargassum polysaccharides calcium complex osteoporosis is as follows:

Make the mouse osteoporosis model with vitamin A acid, the back leg that gavages seven days mouse of vitamin A acid can not stood, can not walk after 12 days, experimental group gavages the sargassan calcium complex, control group gavages the battle array calcium element that Rocaltrol that Switzerland produces and injection Switzerland produce, medication after 15 days the anesthesia of used laboratory animal injection Thiopental Sodium carry out X-photo-beat photograph, observe the pathological change of its bone.The result shows the bone that gavages sargassan calcium complex mouse, turns white in the joint, and is shinny, equally matched with injection thyrocalcitonin group situation, and control group and Rocaltrol group mouse whole body multiple fracture, the artis blackout.After Thiopental Sodium anesthesia, blank group, all large quantities of death of positive drug control group reach 50% as Rocaltrol group dead mouse.And sargassan calcium complex group is all survived, and lives to such an extent that healthy active this explanation sargassan calcium complex is is not only prevented and treated the osteoporosis effect, protects cerebral neural effect in addition.

Below in conjunction with embodiment the present invention is described in detail.Embodiment 1: preparation Sargassum polysaccharides calcium complex

Get the Na that clean dried sargassun 100 g are placed on 0.5wt% concentration ₂CO ₃Soaked 24 hours in the 5000ml solution, become the heavy-gravity dark brown solution, filter residue after filtration, in solution, add the CaCl of 2 mol/L ₂About solution 100 ml, the limit edged stirs, and adds 2 mol/L NaOH solution simultaneously, and the pH value of regulator solution makes its solution keep alkalescence, observes in the solution to add CaCl ₂Solution has no longer added ionic calcium soln till can not dissolving, continuing to stir adding 2 mol/L NaOH stops to add when PH=9-11 scope, restir 0.5 hour, after little precipitation all dissolves like the snowflake, stop to stir the industrial alcohol that adds 2 times of total liquor capacities again, produce a large amount of flockss, place filtration or centrifugal in 3 hours under the room temperature, gained canescence product is the Sargassum polysaccharides calcium complex.Embodiment 2: preparation Sargassum polysaccharides calcium complex

Step adopts the clean dry sargassun 200g and the Na of 5wt% concentration with embodiment 1 ₂CO ₃Be raw material 10000 ml, soaked 12 hours, the ionic calcium soln CaCl of 2mol/L ₂Solution is regulated pH value with 2 mol/L NaOH solution, and the ethanol that adds 2 times of cumulative volumes produces precipitation rapidly, more after filtration, drying obtains final canescence Sargassum polysaccharides calcium complex product.Embodiment 3: preparation Sargassum polysaccharides calcium complex

Press the preparation of embodiment 1 step, wherein raw material adopts clean dried algae sargassun 50 g, Na ₂CO ₃Concentration is to soak 18 hours among 2.5wt% (weight percent) solution 2500 ml, and adding concentration is the CaCl that 2 mol/ rise ₂Calcium solution, and to add NaOH concentration simultaneously be that 2mol/L stirred 1 hour, placed 5 hours under the room temperature, all the other prepare canescence Sargassum polysaccharides calcium network and thing with embodiment 1.Embodiment 4: preparation Sargassum polysaccharides calcium complex

Press embodiment 1 step, condition preparation, just adopt the dried sea-tangle of cleaning to do raw material, get sea-tangle 100g, be placed on the Na of 3wt% concentration ₂CO ₃Soaked 24 hours in the 5000 ml solution, filter residue after filtration, in solution, add the CaCl of 2mol/L ₂About solution 100 ml, the limit edged stirs, and adds the pH value of 2mol/LNaOH regulator solution simultaneously, makes its solution keep alkalescence, observes in the solution to add CaCl ₂Till can not dissolving, solution no longer adds CaCl ₂Solution, continuing to stir adding 2 mol/L NaOH stops to add when PH=9-11 scope, restir 0.5 hour, after little precipitation all dissolves like the snowflake, stop to stir the industrial alcohol that adds 2 times of total liquor capacities again, produce a large amount of flockss, place filtration or centrifugal in 3 hours under the room temperature, gained canescence product is the laminarin calcium complex.Embodiment 5: preparation Sargassum polysaccharides calcium complex

According to step and the reagent dosage of embodiment 4, just adopt the Na of 1.5wt% concentration fully ₂CO ₃Soaked red algae 1000g 20 hours, all the other conditions are constant, make canescence laminarin calcium complex.Embodiment 6: preparation Sargassum polysaccharides calcium complex

The same with embodiment 1 fully, just marine alga is adopted blue algae 200g, with the Na of 4wt% concentration ₂CO ₃Soaked blue algae 20 hours, all the other conditions are identical, make the Sargassum polysaccharides calcium complex.Embodiment 7: preparation Sargassum polysaccharides calcium complex

Preparation method such as embodiment 1, marine alga is adopted chrysophyceae 400g, uses the Na of 2wt% concentration again ₂CO ₃Solution soaking chrysophyceae 24 hours, all the other conditions are identical, are prepared into the Sargassum polysaccharides calcium complex.Embodiment 8: preparation Sargassum polysaccharides calcium complex

Preparation method such as embodiment 1, marine alga is adopted brown alga 300g, uses the Na of 3.5wt% concentration again ₂CO ₃Solution soaking brown alga 22 hours, all the other conditions are identical, are prepared into the Sargassum polysaccharides calcium complex.Embodiment 9: preparation Sargassum polysaccharides calcium complex

Preparation method such as embodiment 1, marine alga is adopted green alga 400g, uses the Na of 2.5wt% concentration again ₂CO ₃Solution soaking green alga 24 hours, all the other conditions are identical, are prepared into the Sargassum polysaccharides calcium complex.Embodiment 10: measure its Plasma Concentration with the S.D rat oral gavage

Used Sargassum polysaccharides calcium complex is that 0.1000g is dissolved in 10 ml tap waters, and solution is creamy white, and does not have precipitation, pH value=8-9.Get 20 of S.D rats, male and female half and half, body weight are 250 grams.Test and ran out of grain in preceding 12 hours, only supply water.Earlier get blood once (excision eyeball) before the administration, get serum and make blank from eye socket.The Sargassum polysaccharides calcium complex is soluble in water, irritates the administration of stomach mode with 300 ml/Kg agent.Get blood half an hour after the administration, 1.5h, and 3.0h, 6.0h respectively get blood once, centrifugal, get serum 0.05ml and add calcon, after the mixing immediately with 722 spectrophotometers in 612nm place colorimetric, be blank with distilled water, zeroing deducts blank colour developing liquid absorbancy, prints the absorbancy for sample.The calculation formula of calcium ion content in the serum: serum calcium mg/dl=

* 10 (reference liquid is 10 milliliters) are slogan banner with the blood sampling time, and the logarithmic value of serum calcium levels draws the curve with Shi Yiliang for vertical mark mapping, as Fig. 1 be oral administration draw the time one discharge curve figure shown in.Eliminate speed Kel=2.3 * B hl=0.02logc=-β t+log c and eliminate the transformation period $t_{1/2\mathrm{\β}}=\frac{\mathrm{Lg}0.5\×2.3}{\mathrm{Kel}}=\frac{0.693}{\mathrm{Kel}}$ $=\frac{0.693}{0.02}=34.65h$ Oral administration belongs to the one-compartment model distribution half-life basically $t_{1/2\mathrm{\α}}=\frac{0.693}{1.28}=0.51h$ Embodiment 11: the experimental result of Sargassum polysaccharides calcium complex bioavailability:

Get 100 of Kunming mouses, body weight 20 ± 2g, male and female half and half are divided 10 groups.

Used medicine is: A. ties up sour potassium and is used to make the horizontal type of osteoporosis.Chongqing pharmaceutical factory produces B. calcitonin treatment osteoporosis specifics.Switzerland Sando z Pharma Inc. produces C. Rocaltrol vitamins D ₃The biological activity medicine.Switzerland Hoffmann company produces D. Calcium of intensifiens Guangzhou emigrant light pharmaceutical factory and produces E. and rely the synthetic F. Sargassum polysaccharides calcium complex oneself of basic acid complex oneself to synthesize

Experimentation: except that the thyrocalcitonin intramuscular injection, all the other are all to irritate the administration of stomach mode with above each medicine.Dosage is tieed up sour potassium once a day, every mouse 2mg.Every mouse intramuscular injection 0.05 ml of positive control drug thyrocalcitonin, three days once.It is 0.5 ml that Rocaltrol is irritated stomach, includes 0.00125 μ g, and per three days once.The every mouse of Calcium of intensifiens mg every days 2.5 is dissolved in the 0.5 ml water L-Lysine calcium salt 0.5 ml.Experiment divides two groups with the Sargassum polysaccharides calcium complex, high dose group 400 mg/Kg, a low dose group 300mg/Kg.

Mouse bone growth situation: test after 12 days, with Thiopental Sodium anesthesia, at Guangzhou Teaching Hospital No.3 of Zhongshan Medical Univ. orthopaedics X-ray sheet of taking pictures.Photographical condition: 33KVP, 1.5 mA, 75 cm, soft rays, 11 slice, thin pieces of taking pictures altogether, the bone growth situation is listed following table in.Current experiment obtains unexpected a discovery, and that is exactly that each group all had death after mouse was anaesthetized with Thiopental Sodium, wherein, L-Lysine calcium salt experimental group and Rocaltrol experimental group mortality ratio are the highest, and the death of Sargassum polysaccharides complexing calcium experimental group is minimum, and concrete numeral please be seen following table: with mouse situation behind the Thiopental Sodium

Group	Original number of elements	Dead	Situation	Skeleton situation *
					Vitamin A acid+Calcium of intensifiens	10	4	Difference	Generally
Dimension+extra large sugared calcium (L)	8	1	Vivaciously	Light in the joint
					Dimension+extra large sugared calcium (H)	8	1	Vivaciously	Light in the joint
Dimension+L-Lysine calcium salt	10	7	Only surplus three on the point of dying	Blackout
					Dimension+thyrocalcitonin	10	3	Difference	Generally
Dimension+Rocaltrol	10	6	Difference	Generally
					Vitamin A acid	10	4	Outage is poor	Blackout
Sargassum polysaccharides calcium (L)	10	1	Vivaciously	It is shinny to turn white
					Sargassum polysaccharides calcium (H)	10	1	Vivaciously	Light in the joint
Contrast	10	2	The difference torpescence	Generally

* skeleton turns white shinnyly for the bone density height, and turning black is osteoporosis.Embodiment 12: the toxicologic result of Sargassum polysaccharides calcium complex:

NIH is that mouse (mix, and divides two groups, and one group is control group, and one group is seaweed polysaccharide trace-element calcium complex experimental group, and two groups feeds utilized identical by 20-30g) 20 of body weight, male and female.With 4.0gkg ^-1Dosage gavage the seaweed polysaccharide trace-element calcium complex to experimental mice.Irritated stomach 15 days, none death.It is no abnormal to observe its outward appearance behavior, and mouse is put to death, and it is no abnormal to observe internal organ, and its heart, liver, kidney are made paraffin section, carries out microscopy at microscopically, and the result does not find any unusual.Only calculating oral maximum tolerated dose is 9.9gk ^-1KD ₅₀Kg ^-1, belong to non-toxic substance.

Claims (6)

1. Sargassum polysaccharides calcium complex is characterized in that:

M is Ca in the formula ²⁺

2. method for preparing the described Sargassum polysaccharides calcium complex of claim l is characterized in that:

1. be in the 0.5-5wt% sodium carbonate solution for the 100g dry seaweed is soaked into 1000-10000ml concentration in proportion, after-filtration extracted polysaccharides in 12-24 hour;

2. be mixed with the CaCl of 2mol/L concentration ₂, solion 80-160ml;

The ionic calcium soln that 3. will prepare 2mol/L concentration is added drop-wise in the polysaccharides that has extracted, the limit edged stirs and carries out complex reaction till solution is saturated, the sodium hydroxide solution that splashes into 2mol/L simultaneously keeps the solution pH value to stop to add alkali in the 9-11 scope, continue to stir 0.5-2 hour, generate Sargassum polysaccharides calcium complex solution;

4. add alcohol in the Sargassum polysaccharides calcium complex solution that 3. step generates, it is 2 times of Sargassum polysaccharides calcium complex liquor capacity total amounts that institute adds pure amount, produces precipitation, after filtration or centrifugation, drying, makes the Sargassum polysaccharides calcium complex.

3. by the method for each Sargassum polysaccharides calcium complex of the described system of claim 2, it is characterized in that: described marine alga comprises: sargassun, brown alga, red algae. blue algae, green alga, chrysophyceae.

4. press the method for the Sargassum polysaccharides calcium complex of the described preparation of claim 2, it is characterized in that: 4. described step makes dried Sargassum polysaccharides calcium complex again through being ground into required particle.

5. the purposes by the described Sargassum polysaccharides calcium complex of claim 1 is characterized in that: as foodstuff additive.

6. purposes by the described Sargassum polysaccharides calcium complex of claim 1 is characterized in that: preparation prevention and treat osteoporosis, treat cardiovascular disorder and protect the purposes of cerebral neural medicine aspect.