CN103622989B - A kind of complex polysaccharide composition and method of making the same and application - Google Patents
A kind of complex polysaccharide composition and method of making the same and application Download PDFInfo
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Abstract
The invention discloses a kind of complex polysaccharide compositions, with Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis for raw material, prepare through water extraction, concentrated, Ethanol Treatment, also disclose the preparation method of above-mentioned complex polysaccharide compositions, and disclose above-mentioned complex polysaccharide compositions and preparing the application that has in the health food of nourishing kidney-yin effect.
Description
Technical field
The invention belongs to polysaccharide technical field, be specifically related to a kind of complex polysaccharide composition and method of making the same and application.
Background technology
The main Physiological Function of kidney yin be promote body moist, quiet, be shaped and restriction sun heat, by internal organs, meridians, body, official's keys of three burnt arrival whole bodies.So kidney yin is vigorous, then whole body the moon is all vigorous; Kidney yin declines few, then whole body the moon all declines; Kidney yin is died, then the moon of whole body is all died, and people is also dead.Deficiency of the kidney yin refers to kidney YIN-fluid being insufficient, is more common in women, and common sympton has soreness of waist and knee joint, dizziness and tinnitus, insomnia and dreamful sleep, dysphoria with feverish sensation in the chest palms and soles, hectic fever night sweat etc.The kidney that the traditional Chinese medical science refers to is relevant with hormone secretion, nervous system, skeleton, reproduction and urinary system, so nourish when kidney portion lacks, will cause the symptom that lassitude loin and legs, menoxenia and urine are a lot of.Mrs climacteric hormone secretion has some changes, and deficiency of the kidney yin situation is even more serious.The traditional Chinese medical science is mainly nursed one's health with Chinese herbal medicine, reaches the object of enriching yin and nourishing kidney, has good curative effect.
At present not yet there is people with Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis for raw material, obtain the composite polysaccharide of each raw material, and for nourishing kidney-yin aspect.
Summary of the invention
First technical problem to be solved by this invention is to provide a kind of complex polysaccharide compositions, and said composition has effect of nourishing kidney-yin.
Second technical problem to be solved by this invention is to provide the preparation method of above-mentioned complex polysaccharide compositions, and this preparation method technique is simple, and extraction efficiency is high.
3rd technical problem to be solved by this invention is to provide above-mentioned complex polysaccharide compositions and is preparing the application had in the health food of nourishing kidney-yin effect.
First technical problem to be solved by this invention is achieved by the following technical solution: a kind of complex polysaccharide compositions, with Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis for raw material, prepares through water extraction, concentrated, Ethanol Treatment.
The weight of each raw material of the present invention is preferably: Radix Rehmanniae Preparata 10 ~ 200, Fructus Lycii 8 ~ 100, Rhizoma Polygonati 8 ~ 100, Radix Codonopsis 4 ~ 50.
The weight of each raw material of the present invention is more preferably: Radix Rehmanniae Preparata 30 ~ 150, Fructus Lycii 40 ~ 80, Rhizoma Polygonati 40 ~ 80, Radix Codonopsis 10 ~ 50.
Weight the best of each raw material of the present invention is: Radix Rehmanniae Preparata 90, Fructus Lycii 60, Rhizoma Polygonati 60, Radix Codonopsis 30.
In complex polysaccharide compositions preparation process of the present invention: during water extraction, during each extraction, the consumption of water is preferably medical material gross weight 5 ~ 20 times, and extract preferably 2 times, each extraction time is 1.5 ~ 4h.When wherein first time extracts, the consumption of water is preferably 10 ~ 20 times of raw material gross weight, boils 2 ~ 4h; When second time is extracted, the consumption of water is preferably 5 ~ 10 times of raw material gross weight, boils 1.5 ~ 3h.
In complex polysaccharide compositions preparation process of the present invention: adopt volumn concentration to be the ethanol water of 70 ~ 95% during Ethanol Treatment.
Multiple condensing mode of the prior art can be adopted time concentrated, wherein best with vacuum concentration.
Second technical problem to be solved by this invention is achieved by the following technical solution: the preparation method of above-mentioned complex polysaccharide compositions, containing following steps: get each raw material of Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis, compare by above-mentioned metering, after mixing, prepare through water extraction, concentrated, Ethanol Treatment.
In above-mentioned preparation method:
During water extraction, the consumption of water is each raw material gross weight 5 ~ 20 times, and extract 2 times, each extraction time is 1.5 ~ 4h; Volumn concentration is adopted to be the ethanol water of 70 ~ 95% during Ethanol Treatment.
3rd technical problem to be solved by this invention is achieved by the following technical solution: above-mentioned complex polysaccharide compositions is preparing the application had in the health food of nourishing kidney-yin effect.
Wherein complex polysaccharide compositions, can be directly oral, also can make said multiple dosage form on pharmaceutics, as capsule-type, tablet, powder, granule or oral liquid etc., has effect of nourishing kidney-yin after edible.
The present invention has the following advantages: complex polysaccharide of the present invention to be rich in the Chinese herbal medicine of active polysaccharide for major ingredient, utilize various polysaccharide component reasonable combination, coordinate share, support kidney by number of ways YIN nourishing, to subhealth state health care there is good effect.
Accompanying drawing explanation
Fig. 1 is the total ion current figure of " deficiency of the kidney yin " rat of hydrocortisone induction in the rat endogenous metabolite change of detailed description of the invention Part II 4.2.1 hydrocortisone induction, gives hydrocortisone continuously 7 days, drug withdrawal the 3rd day serum sample;
Fig. 2 is the total ion current figure of metabolite in normal rabbit serum in the rat endogenous metabolite change of detailed description of the invention Part II 4.2.1 hydrocortisone induction;
Fig. 3 is rat " deficiency of the kidney yin " model group (HC of hydrocortisone induction in the rat endogenous metabolite change of detailed description of the invention Part II 4.2.1 hydrocortisone induction
2) and normal group (N
2) endogenous metabolism thing PCA(principal component analysis);
Fig. 4 is rat " deficiency of the kidney yin " model group (HC of hydrocortisone induction in the rat endogenous metabolite change of detailed description of the invention Part II 4.2.1 hydrocortisone induction
2) and normal group (N
2) endogenous metabolism thing PLS-DA(partial least squares discriminant analysis);
Fig. 5 is the typical total ions chromatogram of intervening metabolite in complex polysaccharide intervention " deficiency of the kidney yin " rat sample rat blood serum during detailed description of the invention Part II 4.2.2 complex polysaccharide changes for endogenous metabolism thing, tests the 22nd day sample;
Fig. 6 is that detailed description of the invention Part II 4.2.2 complex polysaccharide changes the typical total ions chromatogram of metabolite in positives medicine LIUWEI DIHUANG WAN intervention " deficiency of the kidney yin " rat sample rat blood serum for endogenous metabolism thing, tests the 22nd day sample;
Fig. 7 is that detailed description of the invention Part II 4.2.2 complex polysaccharide is for experiment the 22nd day normal group (N in the change of endogenous metabolism thing
2), (complex polysaccharide intervenes rat " deficiency of the kidney yin " model (CPS to pharmaceutical intervention group
2-HC
2) group) and model group (HC
2) endogenous metabolism thing PLS-DA schemes;
Fig. 8 is that detailed description of the invention Part II 4.2.2 complex polysaccharide tests the 22nd day normal group (N for detailed description of the invention Part II in the change of endogenous metabolism thing
2), (LIUWEI DIHUANG WAN intervenes rat " deficiency of the kidney yin " model (PC to pharmaceutical intervention group
2-HC
2) group) and model group (HC
2) endogenous metabolism thing PLS-DA schemes;
Fig. 9 is that detailed description of the invention Part II 4.2.2 complex polysaccharide is analyzed for endogenous metabolism thing change Chinese medicine intervention group and model group endogenous metabolism thing PLS-DA, and wherein pharmaceutical intervention group refers to that complex polysaccharide intervenes rat " deficiency of the kidney yin " model (CPS2-HC2) group;
Figure 10 is that detailed description of the invention Part II 4.2.2 complex polysaccharide is analyzed for endogenous metabolism thing change Chinese medicine intervention group and model group endogenous metabolism thing PLS-DA, and wherein pharmaceutical intervention group refers to that positive drug intervenes rat " deficiency of the kidney yin " model (PC2-HC2) group;
Figure 11 is detailed description of the invention Part II 4.3.3 normal group, model group and the complex polysaccharide intervention group rat blood serum metabolite time dependent PLS-DA trajectory diagram of model group in mutation analysis in time;
Figure 12 is detailed description of the invention Part II 4.3.3 normal group, model group and the complex polysaccharide intervention group rat blood serum metabolite time dependent PLS-DA trajectory diagram of complex polysaccharide group in mutation analysis in time;
Figure 13 is detailed description of the invention Part II 4.3.3 normal group, model group and the complex polysaccharide intervention group rat blood serum metabolite time dependent PLS-DA trajectory diagram of mutation analysis positives medicine group in time.
Detailed description of the invention
The each raw material adopted in following examples, if no special instructions, is commercially available prod.
Part I complex polysaccharide composition and method of making the same
Embodiment 1
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 1 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 90kg, Fructus Lycii 60kg, Rhizoma Polygonati 60kg, Radix Codonopsis 30kg.
Preparation process is: get each raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 10 times of raw material gross weight, boils 2h; When second time is extracted, the consumption of water is 5 times of raw material gross weight, boil 1.5h, filter, merging filtrate, vacuum concentration, carries out Ethanol Treatment by concentrated solution, during Ethanol Treatment, the volumn concentration of ethanol is the ethanol water of 70%, and taking precipitate obtains Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis complex polysaccharide compositions.
Embodiment 2
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 2 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 140kg, Fructus Lycii 40kg, Rhizoma Polygonati 30kg, Radix Codonopsis 10kg.
Preparation process is: get each raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 15 times of raw material gross weight, boils 3h; When second time is extracted, the consumption of water is 8 times of raw material gross weight, boils 2h, and filter, merging filtrate, vacuum concentration, carries out Ethanol Treatment by concentrated solution, and during Ethanol Treatment, the volumn concentration of ethanol is the ethanol water of 80%.
Embodiment 3
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 3 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 50kg, Fructus Lycii 90kg, Rhizoma Polygonati 70kg, Radix Codonopsis 20kg.
Preparation process is: get each raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 20 times of raw material gross weight, boils 4h; When second time is extracted, the consumption of water is 10 times of raw material gross weight, boils 3h, and filter, merging filtrate, vacuum concentration, carries out Ethanol Treatment by concentrated solution, and during Ethanol Treatment, the volumn concentration of ethanol is the ethanol water of 95%.
Embodiment 4
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 4 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 30kg, Fructus Lycii 90kg, Rhizoma Polygonati 90kg, Radix Codonopsis 30kg.
Preparation process is: get each raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 12 times of raw material gross weight, boils 2h; When second time is extracted, the consumption of water is 6 times of raw material gross weight, boils 2.5h, and filter, merging filtrate, vacuum concentration, carries out Ethanol Treatment by concentrated solution, and during Ethanol Treatment, the volumn concentration of ethanol is the ethanol water of 90%.
Embodiment 5
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 5 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 25kg, Fructus Lycii 90kg, Rhizoma Polygonati 80kg, Radix Codonopsis 45kg.
Preparation process is with embodiment 1.
Embodiment 6
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 6 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 50kg, Fructus Lycii 90kg, Rhizoma Polygonati 70kg, Radix Codonopsis 20kg.
Preparation process is with embodiment 2.
Embodiment 7
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 7 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 100kg, Fructus Lycii 60kg, Rhizoma Polygonati 80kg, Radix Codonopsis 15kg.
Preparation process is with embodiment 3.
Embodiment 8
The complex polysaccharide of nourishing kidney-yin effect that what the embodiment of the present invention 8 provided have has the raw material of following weight proportioning to be prepared from: Radix Rehmanniae Preparata 60kg, Fructus Lycii 80kg, Rhizoma Polygonati 80kg, Radix Codonopsis 25kg.
Preparation process is with embodiment 4.
The complex polysaccharide provided by above example is that main composition prepares health food, by common process, can make said multiple dosage form on pharmaceutics, as capsule, tablet, powder, granule or oral liquid etc.
Part II complex polysaccharide compositions has the test of pesticide effectiveness of nourishing kidney-yin effect
Use the proportioning raw materials that above-described embodiment 1 provides, the complex polysaccharide pressed powder (hereinafter referred to as complex polysaccharide) obtained by the technique in embodiment 1 has carried out efficacy validation test, and result of the test is as follows:
1. experiment purpose
Set up the deficiency of the kidney yin rat model of hydrocortisone induction, inquire into the Radix Rehmanniae Preparata in embodiment 1, Fructus Lycii, Rhizoma Polygonati, Radix Codonopsis complex polysaccharide compositions to the impact of deficiency of the kidney yin metabolism of rat group.
2. experiment material
2.1 test specimen
The complex polysaccharide of preparation in embodiment 1, outward appearance is chocolate brown powder, has the distinctive fragrance of this product.
2.2 positive control
Positive control: LIUWEI DIHUANG WAN (water-honeyed pill), the accurate word Z11021283 of traditional Chinese medicines, Beijing Tongrentang Technology Development Co., Ltd. produces.
2.3 reagent and medicine
Injection hydrocortisone sodium succinate (lot number: 20100306), Biochemistry Tianjin Pharmaceutical Co; Chloride injection water, Shanghai ChangZheng Fumin JinShan Pharmaceutical Co., Ltd; Two (trimethylsilyl) trifluoroacetamide (BSTFA, containing 1%TMCS) of heptadecanoic acid, chlorophenylalanine, methoxamine, N-O-, equal purchased from American SIGMA company; Chloroform, methanol, pyridine, be chromatographically pure, purchased from Merck company.
2.4 experimental animal
Male SD rat (180 ~ 200g), 82 (cleaning grade, animal credit number SCXK(Shanghai) 2007-0005), purchased from Shanghai Slac Experimental Animal Co., Ltd..
2.5 experimental apparatus
3. test method
3.1 models are set up and pharmaceutical intervention
The foundation of " deficiency of the kidney yin " model that hydrocortisone (HC) is induced and pharmaceutical intervention experimental design as shown in table 1.
Table 1 experimental design
Note: deficiency of the kidney yin complex polysaccharide 0.26g/kg.d, positive control drug (LIUWEI DIHUANG WAN) 6g/kgd, is all equivalent to 30 times of 60kg adult human dose.
3.1.1 administration and model is prevented to set up
Male SD rat (180 ~ 200g), raises in pharmaceutical college of Shanghai Communications University Experimental Animal Center SPF level barrier animal laboratory.Indoor temperature 20 ~ 25 DEG C, relative humidity 40 ~ 70%, 12 h cycle illuminations, freely ingest, drink water.Animal is fed in bottomless stainless steel silk cylinder mould, 4, every cage.Animal daily nursing and experiment condition all meet " Ministry of Health of the People's Republic of China's laboratory animal environment and facility standard ".
Animal via mated raising after two weeks, was divided into two groups at random, not administration group 18, administration group 40.Administration component is two groups, comprising: positive drug control group (LIUWEI DIHUANG WAN group+blank 10, LIUWEI DIHUANG WAN+modeling group 10); Complex polysaccharide group (complex polysaccharide+blank group 10, complex polysaccharide+modeling group 10).Administration group 12:00 every day gastric infusion, 1 times/day, continuous 15 days, dosage was shown in 3.1.1.The normal saline of not administration group same time every day gavage equivalent volumes.
Test the 16th day, by two administration groups and not administration group respectively stochastic averagina be divided into 2 groups, often organize 9 ~ 10.Get wherein three groups (containing two administration groups and a not administration group) reference literatures and set up deficiency of the kidney yin rat model, in 16:00 right rear leg intramuscular injection every day 25mg/kg 5% hydrocortisone (1mLNS dilute, intramuscular injection volume is: body weight × 10
-3mL), continuous 4 days.Remain the normal saline of three groups of intramuscular injection equivalent volumes.
3.1.2 sample collection
Urine sample: test the 0th, 15,19,22,26,33 day, collects rat 24h urine sample, and record volume of urine, and Quick spin (4 DEG C, 2800rpm), get supernatant subpackage, in-80 DEG C of Refrigerator stores.
Blood sample: test the 0th, 15,22,33 day eye socket and get blood, separation of serum is in-80 DEG C of Refrigerator stores.
3.2 content of observing and indexs
Use ELISA kit to detect each group of purine adenosine phosphate (cAMP) and cyclic guanosine monophosphate (cGMP) level respectively, result represents with mean ± SD.SPSS statistical software (12.0) is adopted to carry out ANOVA analysis to result.
3.3 blood serum metabolic group researchs
3.3.1 serum sample process
Serum dissolves, and after vortex mixing, precision pipettes 100 μ L.Add the heptadecanoic acid 10 μ L of the 1mg/mL of dissolve with methanol and water-soluble 0.3mg/mL chlorophenylalanine 10 μ L, add mixed solvent 300 μ L(chloroform after mixing: methanol 1:3, v/v), vortex mixes,-20 DEG C of standing 10min, 4 DEG C of centrifugal 10min(10,000rpm).Getting 300 μ L supernatant reclaims in sample injection bottle in 1.5mL height, and ambient temperature in vacuum is dry.
Drain rear nitrogen to sample after drying, with ensure in sample anhydrous and in deaeration dampness on derivative impact (time derivative, air humidity≤35%).Add the methoxamine 80 μ L of 15mg/mL pyridinium dissolution after nitrogen dries up, vibrate after sealing 30s, and 30 DEG C of shaking tables (220rpm) react 90min; After reaction terminates, in reaction bulb, add the BSTFA(of 80 μ L containing 1%TMCS), vibrate after sealing 30s, 70 DEG C of reaction 60min.After reaction terminates, vibration 10s, ambient temperatare puts sample introduction analysis after 1h.
3.3.2GC-TOFMS analyze
Serum Silylation thing is analyzed through PegasusHT gas chromatogram-time of-flight mass spectrometer.
Chromatographic condition: chromatographic column DB-5MS capillary column (5%Diphenylcross-linked95%dimethylpolysiloxane:30m × 250 μm i.d., 0.25-μm; AgilentJ & WScientific, Folsom, CA); Splitless injecting samples, sample size 1 μ L; Injector temperature 280 DEG C; Temperature programming condition: initial temperature 80 DEG C; Keep 2min, rise to 180 DEG C with 10 DEG C/min, rise to 230 DEG C with 6 DEG C/min, then rise to 295 DEG C with 40 DEG C/min, keep 8min.Carrier gas is helium (99.9999%), flow rate of carrier gas 1mL/min.
Mass Spectrometry Conditions: solvent time delay 4.5 minutes, ionization mode EI, electron energy-70eV, scanning of the mass spectrum scope 40 ~ 600(m/z); Interface temperature 260 DEG C, ion source temperature 210 DEG C.
3.3.3 date processing and statistical analysis
The initial data that GC-TOFMS instrument detects carries out the pretreatment such as baseline correction, level and smooth and denoising through the software of ChromaTOF and Shanghai Communications University's exploitation and peak extracts and the operation such as peak alignment.Use Excel software and based on MATLABR2010a (MathWorks, Inc.Natick, MA), R10.2 (LucentTechnologies) and JavaSE1.6 (SunMicrosystems) platform Shanghai Communications University exploitation tool kit complete data batch in, batch between correct and Internal standard correction methods.
Identification is carried out in the standard substance storehouse using public database and Shanghai Communications University to set up.Public database mainly comprises NISTlibrary2005, LECO/FiehnMetabolomicsLibraryforGC-TOFMSmetabolomedata (similaritythresholdof70%) etc.; Shanghai Communications University sets up mark product storehouse contains nearly 800 kinds of endogenous metabolism thing.
Use SPSS(SPSS, Chicago, IL), SIMCA-P12.0.1+ (Umetrics,
sweden), Excel(Microsoft, USA), R(GNU) and Shanghai Communications University exploitation software kit etc. one-dimensional and multidimensional statistics analysis are carried out to data.
4. experimental result
4.1 Analysis of Biochemical
Table 2 is group rat urine and Biochemical Indices In Serum analysis respectively
Note: § and blank group ratio, P < 0.05; § § and blank group ratio, P < 0.01;
* with model group ratio, P < 0.05; * and model group ratio, P < 0.01.
Blood plasma cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) are two mutual antagonisms, the mutual neurotransmitteies restricted in cell, and cellular function is in steady statue and has double-direction control regulating action.Root according to research reports, be it is generally acknowledged " syndrome of deficiency of kidney yin ", and cAMP content raises, and cGMP content reduces, thus causes the rising of cAMP/cGMP ratio.To respectively group rat blood serum cAMP and the display of cGMP assay in the 25th day, model group cAMP level is higher than higher than normal group, and cGMP level is lower than normal group, and cAMP/cGMP is also than normal rising, be consistent (table 2) with the performance of bibliographical information yin deficiency syndrome, show the successful of modeling.
As seen from the results in Table 2, compared with model group, complex polysaccharide and the change of positive drug LIUWEI DIHUANG WAN to the rat These parameters that HC induces all have preventive effect to a certain degree.
4.2 metabonomic analysis
4.2.1 the rat endogenous metabolite change of hydrocortisone induction
Adopt the blood serum metabolic group science study method based on TMS derivatization and gas chromatogram-ionization time of flight, " deficiency of the kidney yin " rat of hydrocortisone induction is analyzed with blank group rat blood serum, obtain the total ions chromatogram of extract as depicted in figs. 1 and 2, the intensity of some metabolite therefrom can obviously found out between two groups presents difference.
Change for investigating the metabolic pathway relevant to " deficiency of the kidney yin " that hydrocortisone is induced in serum, carry out PCA and PLS-DA analyze giving the model group of hydrocortisone after 4 days when 3 days and normal rats serum protein moteblites modal data continuously, the results are shown in Figure 3 and Fig. 4.As can be seen from Fig. 3 and Fig. 4, model group and normal group have certain separation trend on PCA figure, but separating effect is unsatisfactory; Two groups can be separated completely on PLS-DA model, and this model has good explanation rate and prediction rate, and model parameter is in table 3.
Table 3 main models parameter
| Sequence number | Model | Composition a | R 2X b | R 2Y c | Q 2Y d | Corresponding diagram |
| 1 | PCA | 2 | 0.423 | ― | 0.023 | Fig. 3 |
| 2 | PLS-DA | 4 | 0.551 | 0.993 | 0.512 | Fig. 4 |
Note: a: model composition number; B: model is to the interpretability of initial data (be extracted the percentage ratio of the quantity of information in data square); C: model is to the capability of fitting instructing variable; D: the predictive ability of modeling.
Based on model group rats and normal rats blood-serum P LS-DA analysis result, this experiment have found 105 Differential variable (VIP>1) altogether as potential metabolic marker thing between two groups.Compose storehouse contrast by composing the self-built standard substance in storehouse (as Willy and NIST mass spectral database) and Shanghai Communications University with the commercialization of generally acknowledging, qualification wherein 42 compounds altogether, in conjunction with T check analysis (P<0.05), final screening obtains 10 diversity metabolite, the results are shown in Table 4.
Difference metabolite in table 4 " syndrome of deficiency of kidney yin " model group and normal rats serum
Note: the retention time that a. metabolite GC-TOFMS analyzes; The variable weight value (Variableimportanceintheprojection) that b.PLS-DA model provides; C.T checks significance; D. in ↓ representative model group, the content compared with normal group of this metabolite is low.
From the results shown in Table 4, the closely-related metabolite of " deficiency of the kidney yin " model of inducing with hydrocortisone relates generally to carbohydrate metabolism, fatty acid metabolism and amino acid metabolism etc.
4.2.2 complex polysaccharide intervention that endogenous metabolism thing is changed
This experimentation complex polysaccharide and positive drug LIUWEI DIHUANG WAN are to the intervention effect of rat " syndrome of deficiency of kidney yin " caused by hydrocortisone, Fig. 5 and 6 is respectively the rat modeling in 4 days that prevention gives complex polysaccharide and positive drug LIUWEI DIHUANG WAN, and drug withdrawal is to serum extract total ions chromatogram time modeling agent the 3rd day (namely testing the 22nd day).
Adopt multidimensional statistics analytical method to carry out PLS-DA modeling analysis to normal group, model group and pharmaceutical intervention group during experiment the 22nd day, the results are shown in Figure 5 and 6, main models parameter is in table 5.
Table 5 main models parameter 1
| Sequence number | Group | Composition a | R 2X b | R 2Y c | Q 2Y d | Corresponding diagram |
| 1 | CPS 2-HC 2,HC 2,N2 | 7 | 0.64 | 0.996 | 0.0272 | Fig. 7 |
| 2 | PC 2-HC 2,HC 2,N2 | 6 | 0.617 | 0.941 | 0.301 | Fig. 8 |
Table 6 main models parameter 2
| Sequence number | Group | Composition a | R 2X b | R 2Y c | Q 2Y d | Corresponding diagram |
| 2 | CPS 2-HC 2,HC 2 | 5 | 0.621 | 0.993 | 0.439 | Fig. 9 |
| 3 | PC 2,HC 2 | 5 | 0.589 | 0.999 | 0.705 | Figure 10 |
Note: a: model composition number; B: model is to the interpretability of initial data (be extracted the percentage ratio of the quantity of information in data square);
c: model is to the capability of fitting instructing variable; D: the predictive ability of modeling.
10 diversity metabolite between table 7 model group and normal group are in the situation of change of Chinese medicine intervention group
Note: a. Metabolites in serum intensity represents with peak area; B. the T of Kidney-yin-vacuity group/pharmaceutical intervention group and normal group checks difference, and * represents P<0.05, and * * represents P<0.01; C. the T of pharmaceutical intervention group and normal group checks difference.
By the metabolic patterns difference of comparative experiments the 22nd day model group and pharmaceutical intervention group, complex polysaccharide group, LIUWEI DIHUANG WAN group and model group are carried out PLS-DA analysis respectively, found that, each administration group and model group all have obvious separation trend, and model explanation rate and prediction rate are shown in Fig. 7-8 and table 6.On this basis, 10 endogenous diversity metabolite in further observing and nursing group and normal group and correlative metabolites are in the situation of change of each pharmaceutical intervention group, and the diversity of these difference things between each pharmaceutical intervention group and normal group is carried out T check analysis, the overwhelming majority all no longer includes without significance (P>0.05), in table 7.This result shows that the change of each administration group to the endogenous metabolism caused due to " syndrome of deficiency of kidney yin " has certain anti-preact, makes the fluctuation of these metabolite levels caused by the deficiency of YIN become no longer obvious.And the change of other correlative metabolites in each administration group is in table 8, the relative peak area intensity of the diversity metabolite as can be seen from the table between absolutely large number model group and normal group is under the intervention of complex polysaccharide and positive control drug, the level of metabolite in normal group is in various degree close, points out each administration group also to have preventive effect to a certain degree to the change of these metabolite.These results suggest that it is that complex polysaccharide or LIUWEI DIHUANG WAN all may have preventive effect to a certain degree to hydrocortisone induction " syndrome of deficiency of kidney yin ".
Other correlative metabolites of table 8 is at the situation of change a of normal group, model group and Chinese medicine intervention group
Note: a. Metabolites in serum intensity is with peak area table
4.2.3 normal group, model group and complex polysaccharide intervention group rat blood serum metabolite mutation analysis in time
By PLS-DA model depiction, model group and two administration group rats are in experiment the 0th day, the 22nd day and metabolism in the 33rd day spectrum variation track figure (Figure 11,12,13).Result shows, and model group presents obvious time dependent track, and metabolic patterns appearance compared with the 0th day of the 22nd day is significantly separated (Figure 11), metabolic regulation network when showing the 22nd day and modeling be front occur significantly different.Compare with model group, after the modeling of two administration groups terminates, the 3rd day (testing the 22nd day) metabolic patterns obviously weakens (Figure 12 and 13) than being separated between model group with normal group with the separation case before modeling, shows the disorder that prevention gives complex polysaccharide or LIUWEI DIHUANG WAN and can prevent the metabolism network that " syndrome of deficiency of kidney yin " causes caused by hydrocortisone to a certain extent.
Based on model group test the 22nd day with the 33rd day metabolite situation of change of relative 0th day, by one-dimensional analysis (T inspection), screen and identify wherein 6 diversity metabolite, calculate the change multiple of these 6 difference metabolite when the 22nd day and 33 days respectively, result shows that these diversity metabolite are very not large from situation of change during stopping modeling the 3rd day to 14 days, point out not repaired completely, in table 9 of corresponding metabolic patterns.
Table 9 model group in time difference in change the opposite sex metabolite significance analysis
Wherein model group-D22 refers to the model group of the 22nd day, and model group-D33 refers to the model group of the 33rd article.
The change multiple of 6 diversity metabolite more than when relative 0th day of experiment the 22nd day and the 33rd day each administration group is analyzed simultaneously, and carry out T inspection (table 10-11).The change of result display complex polysaccharide and LIUWEI DIHUANG WAN group part metabolite 22 days time does not have model group obvious, points out each administration group to present preventive effect to a certain degree to the metabolism network disorder caused by " yin deficiency syndrome ".
A table 106 diversity material time dependent significance analysis (a) in each administration group
Wherein complex polysaccharide-D22 refers to the significance analysis of the 22nd day complex polysaccharide, and complex polysaccharide-D33 refers to the significance analysis of the 33rd day polysaccharide.
A table 116 diversity material time dependent significance analysis (b) in each administration group
Wherein, wherein positive drug-D22 refers to the significance analysis of the 22nd day positive drug, and complex polysaccharide-D33 refers to the significance analysis of the 33rd day positive drug.
By the change of endogenous small molecule metabolites in comparison model group and Chinese medicine intervention group, we find that the change of the above-mentioned metabolism network caused by " syndrome of deficiency of kidney yin " can be resisted or prevent to this complex polysaccharide to a certain extent, the listed disorderly metabolite level of his-and-hers watches 7, table 8 and table 9 has regulating action in various degree, make metabolite metaboilic level return to normal level, draw close, the prevention intervention fully disclosing this complex polysaccharide effectively can prevent the metabolism disorder of hydrocortisone caused " syndrome of deficiency of kidney yin ".
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included in protection scope of the present invention.
Claims (9)
1. a complex polysaccharide compositions, it is characterized in that: with Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis for raw material, prepare through water extraction, concentrated, Ethanol Treatment, the weight of each raw material is: Radix Rehmanniae Preparata 10 ~ 200, Fructus Lycii 8 ~ 100, Rhizoma Polygonati 8 ~ 100, Radix Codonopsis 4 ~ 50.
2. complex polysaccharide compositions according to claim 1, is characterized in that the weight of each raw material is: Radix Rehmanniae Preparata 30 ~ 150, Fructus Lycii 40 ~ 80, Rhizoma Polygonati 40 ~ 80, Radix Codonopsis 10 ~ 50.
3. complex polysaccharide compositions according to claim 2, is characterized in that the weight of each raw material is: Radix Rehmanniae Preparata 90, Fructus Lycii 60, Rhizoma Polygonati 60, Radix Codonopsis 30.
4. the complex polysaccharide compositions according to any one of claim 1-3, is characterized in that: during water extraction, and the consumption of water is 5 ~ 20 times of medical material gross weight, and extract 1 ~ 3 time, each extraction time is 1.5 ~ 4h.
5. the complex polysaccharide compositions according to any one of claim 1-3, is characterized in that: adopt volumn concentration to be the ethanol water of 70 ~ 95% during Ethanol Treatment.
6. the preparation method of the complex polysaccharide compositions described in any one of claim 1-3, is characterized in that containing following steps: get each raw material of Radix Rehmanniae Preparata, Fructus Lycii, Rhizoma Polygonati and Radix Codonopsis, by above-mentioned metering ratio, after mixing, prepares through water extraction, concentrated, Ethanol Treatment.
7. the preparation method of complex polysaccharide compositions according to claim 6, is characterized in that: during water extraction, and the consumption of water is each raw material gross weight 5 ~ 20 times, and extract 1 ~ 3 time, each extraction time is 1.5 ~ 4h; Volumn concentration is adopted to be the ethanol water of 70 ~ 95% during Ethanol Treatment.
8. the complex polysaccharide compositions described in any one of claim 1-3 is preparing the application had in the health food of nourishing kidney-yin effect.
9. complex polysaccharide compositions according to claim 8 is preparing the application had in the health food of nourishing kidney-yin effect, it is characterized in that: the dosage form of described health food is capsule-type, tablet, powder, granule or oral liquid.
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