CN1277027A - Silkworm excrement extracting process, extract and its use - Google Patents
Silkworm excrement extracting process, extract and its use Download PDFInfo
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Abstract
Silkworm excrement extracting process includes continuous or intermittent extraction with solvent in the amount of 4-14 times that of medicine material, filtering to eliminate sludge, heating to concentrate, cooling, addition of alcohol, filtering to eliminate insoluble matter, merging filtrate, further concentration into ointment and forming. The extracted matter is used to prevent and treat diabetes.
Description
The present invention relates to extracting method, extract and the new purposes thereof of Chinese medicine Bombyx mori L..
In motherland's traditional medicine, Bombyx mori L. (Faeces bombycis) is the granule of short cylindrical shape, long 2-5mm, diameter 1.5-3mm.The surface grey black is to green black, and is coarse, and 6 vertical ribs and horizontal ring grain are arranged, and two ends are blunt, are six prismatics, and matter is hard and crisp.Tool grass gas, lightly seasoned.Has expelling wind and removing dampness, the effect of the analgesic therapy of invigorating blood circulation.Be used for rheumatic arthralgia, unsuccessful, the munbness in joint, waist lower limb cold type of pain, rubella pruritus; The wind syndrome of head headache, baking string wind-eye.Prior art bibliographical information Bombyx mori L. crude drug contains chlorophyll, pohytol, carotenoid, vitamin A, compositions such as B and free amino acid.Also contain compositions such as polysaccharide, monosaccharide, vegetable protein, phenols, flavonoid, organic acid, aminoacid in this medical material.
The object of the present invention is to provide a kind of Bombyx mori L. extract.
Another object of the present invention provides a kind of extracting method that extracts Bombyx mori L..
A further object of the present invention provides a kind of new purposes of extract of the present invention.This extract has the effect of the blood glucose that reduces mammal and people.
Have with the different effect of prior art owing to adopt the present invention's method to extract the product that obtains, make the Bombyx mori L. extract have medicine and industrial use widely, for treatment and prevent diabetes provide a new alternative approach.
In order to finish the present invention's above-mentioned purpose, the concrete technical scheme that adopts of the present invention is:
The extracting method of Bombyx mori L. extract of the present invention (hereinafter to be referred as FB) may further comprise the steps:
A crude drug water is static or dynamically down, room temperature or heating (or backflow) continuously or intermittent extraction, the 4-14 that extracts quantity of solvent and be former medicine weight is doubly;
B elimination medicinal residues, filtrate are heated to 40-100 ℃ and are concentrated into medical material weight 1-5 volume postcooling doubly under normal pressure or decompression, dynamical state; Heating-up temperature is preferably 60-65 ℃
C adds alcohol then makes it become the concentration of 40-80%, removes by filter insoluble matter, filtrate merging further is condensed into paste carries out preparations shaping.
Can repeat 1-4 time when adopting the intermittent extraction step.Described alcohol comprises alcohol such as methanol well known to those skilled in the art, ethanol, isopropyl alcohol, butanols.
Another extracting method two of the present invention may further comprise the steps:
The A crude drug uses alcohols (methanol, ethanol, isopropyl alcohol, butanols etc.) or aqueous alcohol (40-80%) in static state or down dynamic, and 40-100 ℃ of extraction of room temperature or heating or reflux, extract,, extraction quantity of solvent are 4-14 times of former medicine weight;
B elimination medicinal residues, filtrate are concentrated under the medical material weight 1-5 volume doubly in normal pressure or decompression, 40-100 ℃ of heating under the dynamical state and cool off;
The centrifugal insoluble matter of removing of C, and add a small amount of washing insoluble matter 1-3 time, merging filtrate further is condensed into paste and carries out preparations shaping;
D extractum can become dry powder through the direct spray drying of concentrated liquid, lyophilization or vacuum drying, and preparations shaping.
Extraction can be carried out continuously or intermittently, can repeat 1-4 time during intermittent extraction,
After the extracting solution of method one and method two can adopt the method for well known to a person skilled in the art directly to use exchange column (comprising exchange columns such as macroporous resin, ion exchange resin, polydextran gel) and membrane technology to make with extra care again to concentrate, become extractum or dry powder again.
The inventor has blood-sugar decreasing active through pharmacological evaluation proof Bombyx mori L. extract.Continue the pharmacological action of its active component of research, can develop the medicine of more effective blood sugar lowering or other purposes.
Describe the present invention in detail below with reference to embodiment.
Embodiment 1
Get 1 kilogram of crude drug Bombyx mori L., add 4 times of amount 60% ethanol, soaked overnight or 4 hours slowly are heated to boiling, reflux filtration 1 hour; Overall process repeats once, merging filtrate, concentrating under reduced pressure, 40 ℃ of temperature, pressure 85-98KPa, be concentrated into the water that adds when not having ethanol fully with the medical material identical weight and continue to be heated to boiling, the centrifugal insoluble matter of removing in cooling back, and add a small amount of washing insoluble matter three times, filtrate is merged, at temperature 60-70 ℃ of following concentrating under reduced pressure, drying under reduced pressure (80 ℃ of temperature, pressure 98Kpa) gets dry powder then.Extraction rate 5-7%.
Embodiment 2
Get 1 kilogram of crude drug Bombyx mori L., add 14 times of amount methanol, soaked overnight slowly is heated to boiling, refluxes 3 hours, filters; Overall process repeats four times, merging filtrate, concentrating under reduced pressure, 100 ℃ of temperature, pressure 98KPa, be concentrated into the water that adds when not having ethanol fully with the medical material identical weight and continue to be heated to boiling, the centrifugal insoluble matter of removing in cooling back, and add a small amount of washing insoluble matter three times, filtrate is merged, at 60 ℃ of following concentrating under reduced pressure of temperature, drying under reduced pressure (70 ℃ of temperature, pressure 85Kpa) gets dry powder then.Extraction rate 5-7%.
Embodiment 3
Get 1 kilogram of crude drug, the water logging bubble that adds 4 times of amounts spends the night, and slowly is heated to boiling, continued reflux, extract, 1 hour, and filtered then, overall process repeats four times, merging filtrate, concentrating under reduced pressure, 60 ℃ of temperature, pressure is 80KPa, is concentrated into the volume postcooling of 1 times of medical material weight, adds ethanol then, makes it become 60% concentration, placed 4 hours, and removed by filter insoluble matter, merging filtrate, further concentrating under reduced pressure under 60 ℃, dry (temperature 60-70 ℃) gets dry powder then.Extraction ratio is 8.8%.
Embodiment 4 gets 1 kilogram of crude drug, the water logging bubble that adding 14-doubly measures spends the night, slowly be heated to boiling, continued reflux, extract, 1 hour, filter then, overall process repeats once, merging filtrate, concentrating under reduced pressure, 70 ℃ of temperature, pressure is 85-98KPa, is concentrated into the volume postcooling of 1 times of medical material weight, add 95% ethanol then, make it become 60% concentration, placed 4 hours, and removed by filter insoluble matter, merging filtrate, further concentrating under reduced pressure under 50 ℃, dry (temperature 60-70 ℃) gets dry powder then.Extraction ratio is 7.2%.
The main pharmacodynamics of Bombyx mori L. extract for treating diabetic can be obtained a result by following experiment:
Male Kunming strain mice, body weight 22-25g No. the 029th, capital moving pipe matter word (1994) (the animal quality quality certification number for), the wistar rat, male, body weight 180-250g No. the 030th, capital moving pipe matter word (1994) (the animal quality quality certification number by) is all bred factory available from Chinese Academy of Medical Sciences's zooscopy.Alloxan induced hyperglycemia mice, rat model: give intact animal's intravenous injection alloxan (mice 90100mg/kg, rat 45-50mg/kg), blood glucose (glucose oxidase method) is predicted in administration after 72 hours, select for use blood glucose value to experimentize the above person of 11.1mmol/L.
Test example 1
The Bombyx mori L. extract is to the influence of alpha-glucosidase activity
With the alpha-glucosidase of dog or rat preduodenal, measure the inhibitory action of the FB of variable concentrations, IC to saccharase and maltase activity
50Be respectively 6.8ug/ml and 10.3ug/ml.Illustrate that FB has the enzyme inhibition of α glucoside.
Test example 2
The Bombyx mori L. extract to alloxan induced hyperglycemia mice sucrose load and starch-bearing after the influence of blood sugar increasing
Two batches of alloxan induced hyperglycemia mices, 4 groups every batch, every group 10, test overnight fasting on the same day, irritate stomach with sucrose (4.0g/kg) or starch (3.0g/kg) for one group and organize in contrast, all the other groups are irritated FB (0.6g/kg, the 1.2g/kg of stomach sucrose or starch and various dose respectively, 1.8g/kg), the blood glucose value (table 1,2) when measuring 30min, 60min after 0min and the administration, 120min.When table 1. Bombyx mori L. extract is loaded back glycemic peaks, peak value to alloxan induced hyperglycemia mice sucrose
Between and influence (mean ± SD, n=10) the group glycemic peaks time to peak AUC of area under curve
(mmol/L) (min.) (mmol/L.hr) matched group 22.6 ± 1.7 30 39.0 ± 4.2FB 0.6 19.3 ± 2.7** 60 34.2 ± 4.8*FB 1.2 17.1 ± 1.3*** 60 30.8 ± 2.8***FB 1.8 14.8 ± 1.8*** 60 28.2 ± 4.8***AUC is an area under the blood glucose curve; Compare * P＜0.05, * * P＜0.01, * * * p＜0.001 with matched group; FB dosage be g/kg table 2. Bombyx mori L. extract to the alloxan induced hyperglycemia mice starch-bearing after glycemic peaks, peak value
The influence of time and area under curve (mean ± SD, n=10) group glycemic peaks time to peak AUC
(mmol/L) (min.) (mmol/L.hr) matched group 24.4 ± 1.3 30 40.6 ± 4.4FB 0.6 23.0 ± 2.9 60 36.6 ± 5.9FB 1.2 21.3 ± 3.1** 60 35.9 ± 4.6*FB 1.8 19.3 ± 2.2*** 60 32.9 ± 3.9***AUC is an area under the blood glucose curve; Compare * P＜0.05, * * P＜0.01, * * * p＜0.001 with matched group; FB dosage is g/kg
Presentation of results, the FB of various dose can suppress the rising of blood glucose behind alloxan induced hyperglycemia mice sucrose load and the starch-bearing, and area under the minimizing blood glucose curve moves after making glycemic peaks.
Test example 3
The Bombyx mori L. extract to the normal mouse glucose load after the influence of blood sugar increasing
3 groups of normal mouses are irritated stomach with glucose (4.0g/kg) solution for one group and are organized in contrast, all the other groups irritate respectively stomach glucose and various dose FB (0.45g/kg, 1.8g/kg), the blood glucose value after mensuration 0min and the administration when 30min, 60min, 120min.Administration group and matched group compare, and the equal no difference of science of statistics of area under glycemic peaks, time to peak and the blood glucose curve illustrates that FB brings into play drug action by suppressing alpha-glucosidase, and the not directly absorption of affecting glucose.
Test example 4
The Bombyx mori L. extract is to the influence of alloxan hyperglycemic rat blood glucose and glucose in urine
Three groups of alloxan hyperglycemic rats, every group 10, first group of feed high-sucrose feedstuff contrasts as hyperglycemia, second and third group is taken food respectively, and (amount to dosage is 0.6g/kg to the high-sucrose feedstuff that contains various dose FB, 1.2g/kg), the 4th group is that the normal feedstuff of normal rat (10) feed is as the normal control group.Measure blood glucose after two weeks and collect 6 hours glucose in urine content of urine survey, also collect the 4th group of normal rat urine simultaneously and measure, as a comparison, the results are shown in Table 3 with metabolic cage.
Table 3. Bombyx mori L. extract is to the influence of alloxan hyperglycemic rat blood glucose, glucose in urine
(mean ± SD, n=10) group blood glucose (mmol/L) glucose in urine
Not fasting fasting (g/6hrs) normal rat 5.4+0.2 3.4 ± 0.5 0 hyperglycemic rat 25.9+3.6 16.5+2.1 2.59+0.76FB 0.6 19.1+3.4***, 10.4 ± 3.5*** 0.70+0.15***FB 1.2 10.9 ± 4.4***, 8.3 ± 3.7*** 0.38+0.22*** and hyperglycemic rat group are relatively, * * P＜0.001 presentation of results, FB can obviously reduce the blood sugar level and the glucose in urine content of alloxan hyperglycemic rat.
Test example 5
The Bombyx mori L. extract is to the influence of healthy volunteer's steamed bread test back blood sugar increasing
7 of healthy volunteers (this institute staff), 2 of male, 5 of women, the age is 25-62 year.Test for the first time, early morning, everyone ate 100g steamed bread (this institute dining room provides) as own control on an empty stomach; FB 1.5g/ people (4 people) is taken in test for the second time at random, and 3.0g/ people (3 people) and 100g steamed bread be with clothes, and the blood glucose value when measuring 30min, 60min behind (0min) and food steamed bread on an empty stomach, 120min.Twice experiment be 1 week at interval, the results are shown in Table 4.
Table 4. Bombyx mori L. extract is to healthy volunteer's steamed bread test back glycemic peaks, time to peak
With the influence of area under curve (mean ± SD)
Heavy dose of low dose of
(3.0g/ people) (1.5g/ people)
Before the administration after the administration before the administration after the administration N 3344 glycemic peaks 6.8 ± 1.4 4.6 ± 0.5* 6.5 ± 1.5 5.4 ± 0.9* (mmol/L) time to peak 30 60 30 60 (min.) AUC 12.2 ± 1.9 8.8 ± 0.5* 11.5 ± 3.1 10.0 ± 1.2 (mmol/L.hr) AUC be area under the blood glucose curve; Compare * P＜0.05, * P ≈ 0.05 before and after self
PRELIMINARY RESULTS explanation, FB can make normal person's steamed bread test back glycemic peaks reduce, and reduces area under the blood glucose curve, moves after making peak value.
Test example 6
The agent of Bombyx mori L. extract particles is to the influence of type 2 diabetes mellitus trial volunteer blood glucose
Type 2 diabetes mellitus trial volunteer 10 people that made a definite diagnosis, the age is between 43-83 year, male 7 people wherein, women 3 people, suffering from the diabetes time is 1-7.Take medicine before measurement fasting glucose and steamed bread (100g) load back 1h and 2h blood glucose, after three days, measure the steamed bread load back 1h once obey FB and 2h blood glucose and be mensuration first, take FB afterwards continuously, according to conditions of patients, take medicine every day 2-3 time (2.0g/ time), take after mixing it with water when advising on the feed.Take medicine 2 weeks of back, measure fasting glucose 4 weeks respectively and add steamed bread load back 1h and the 2h blood glucose of FB.Part experimenter has also carried out said determination during 8 weeks.In process on probation, advise the patient to continue to take the former medicine of using, other antidiabetic medicines but individual patient has been stopped using voluntarily.3 patients are arranged because other reasons fails to obtain whole experimental datas according to design on probation, existing with result on trial brief summary following (table 5):
Table 5.2 type diabetes trial volunteer is taken the Bombyx mori L. extract
The front and back change of blood sugar relatively
Pairing t-test#P ≈ 0.05 * P＜0.05 * * P＜0.01 before and after statistical method adopts and takes medicine
| Index | Before taking medicine (n=10) | Take FB | |||
| (n=8) first | 2 weeks (n=8) | 4 weeks (n=7) | 8 weeks (n=6) | ||
| Fasting blood sugar (mg/dl) | 183±65 | ?180±66 | 141±52* | ??146±54* | ??131±61* |
| 1h blood glucose value (mg/dl) behind the bread meal | 273±64 | ?219±67** | 207±67* | ??200±88* | ??192±89# |
| 2h blood glucose value (mg/dl) behind the bread meal | 310±88 | ?287±76 | 247±87* | ??223±88* | ??204±92* |
| Area under the blood glucose curve (mg/dlh) | 514±130 | ?452±138** | 402±136** | ??386±167* | ??361±167* |
As seen from the above table: the 1h blood glucose value obviously reduced after the Bombyx mori L. extract can make type 2 diabetes mellitus trial volunteer steamed bread load, and relatively promptly there were significant differences before taking and take 2 weeks first and trying out, and reduce with the Time of Administration prolongation; Steamed bread load back 2h blood glucose value when taking the Bombyx mori L. extract first with on probation before no significant difference, take 2 week the back obviously reduce, have afterwards and reduce trend gradually, 4 weeks and 8 week the back with try out before significant difference is more also arranged.Fasting glucose reduces gradually after taking the Bombyx mori L. extract, and more all there were significant differences after 2 weeks, 4 weeks and 8 weeks and before on probation.Area relatively has the difference of highly significant under the blood glucose curve when taking medicine first with before taking medicine, and along with prolonging medicine time and reducing gradually.Period in a medicine has patient's hunger sensation of symptom more than three, thirsty symptom obviously to alleviate, and amount of drinking water and urine amount significantly reduce, and all patients are conscious, and muscle power obviously strengthens, and the 2-3 hour after the meal glucose in urine of testing oneself obviously reduces or feminine gender.
Above-mentioned result of the test shows that Bombyx mori L. extract (FB) has the alpha-glucosaccharase enzyme inhibition, can obviously suppress the blood sugar increasing behind diabetic mice sucrose, the starch-bearing, makes that area obviously reduces under glycemic peaks, the blood glucose curve, and moves after making time to peak; Multiple dosing can make alloxan diabetes rats fasting glucose, post-prandial glycemia, glucose in urine be starkly lower than matched group; Normal person's steamed bread test back glycemic peaks is reduced, and reduce area under the blood glucose curve, move after making peak value; The agent of Bombyx mori L. extract particles can make type 2 diabetes mellitus trial volunteer steamed bread tested back 1 hour, 2 hours and blood glucose curve under area obviously reduce, fasting glucose is descended gradually.Therefore, the Bombyx mori L. extract can be used for treatment of diabetes.
Test example 7 acute toxicities
The Bombyx mori L. extract is made into 0.45g/ml, divides 2 times gastric infusion, each 0.2ml/10g body weight, two minor ticks 5 hours.Total dosage is 18.0g/kg (amounting to crude drug 300g/kg).Observed 7 days continuously, do not see animal dead and other poisoning symptom.Cut open inspection after 7 days, also no abnormality seen.
Test example 8 chronic toxicities
The wistar rat is carried out the chronic toxicity test in 24 weeks, observed growth and dead.Respectively at 6 weeks after the 12nd week of administration, the 24th week and the drug withdrawal putting to death the part animal, make routine blood test and blood biochemical regulating liver-QI kidney function test, get main organs row pathologic finding, check result shows: the every inspection of Bombyx mori L. extract treated animal all in normal range, is compared no significant difference with matched group.Illustrate that the Bombyx mori L. extract is safe in utilization.The people estimates dosage Bombyx mori L. extract 1-3g/ time, three times on the one.
Claims (10)
1. Bombyx mori L. preparation method of extract may further comprise the steps:
A Bombyx mori L. crude drug water is static or dynamically down, room temperature or heating (or backflow) continuously or intermittent extraction, the 4-14 that extracts quantity of solvent and be former medicine weight is doubly;
B elimination medicinal residues, filtrate are heated to 40-100 ℃ and are concentrated into medical material weight 1-5 volume postcooling doubly under normal pressure or decompression, dynamical state;
C adds alcohol then makes it become the concentration of 40-80%, removes by filter insoluble matter, filtrate merging further is condensed into paste carries out preparations shaping.
2. the method for claim 1 is characterized in that heating-up temperature is preferably 60-65 ℃.
3. method as claimed in claim 1 or 2 can repeat 1-4 time when it is characterized in that adopting the intermittent extraction step
4. Bombyx mori L. preparation method of extract may further comprise the steps:
A Bombyx mori L. crude drug water or aqueous alcohol (40-80%) are in static state or down dynamic, and room temperature or heating are extracted or reflux, extract, for 40-100 ℃, and the extraction quantity of solvent is 4-14 a times of former medicine weight;
B elimination medicinal residues, filtrate are concentrated under the medical material weight 1-5 volume doubly in normal pressure or decompression, 40-100 ℃ of heating under the dynamical state and cool off;
The centrifugal insoluble matter of removing of C, and add a small amount of washing insoluble matter 1-3 time, merging filtrate further is condensed into extractum and carries out preparations shaping;
5. method as claimed in claim 4 can repeat 1-4 time when it is characterized in that adopting the intermittent extraction step
6. method as claimed in claim 4 is characterized in that extractum can become dry powder through the direct spray drying of concentrated liquid, lyophilization or vacuum drying.
7. method as claimed in claim 4, it is characterized in that can be directly with exchange column (comprising exchange columns such as macroporous resin, ion exchange resin, polydextran gel) and membrane technology make with extra care concentrate after, become extractum or dry powder again.
8. extract the Bombyx mori L. extract that obtains through the described method of claim 1.
9. extract the Bombyx mori L. extract that obtains through the described method of claim 4.
10. Bombyx mori L. or Bombyx mori L. extract are in the application of preparation treatment and prevention mammal and people's diabetes medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991091116A CN1152692C (en) | 1999-06-14 | 1999-06-14 | Silkworm excrement extracting process, extract and its use |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991091116A CN1152692C (en) | 1999-06-14 | 1999-06-14 | Silkworm excrement extracting process, extract and its use |
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| Publication Number | Publication Date |
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| CN1277027A true CN1277027A (en) | 2000-12-20 |
| CN1152692C CN1152692C (en) | 2004-06-09 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100390176C (en) * | 2004-10-12 | 2008-05-28 | 大连理工大学 | Method for extracting chlorophyl from silkworm faeces by microwave pretreatment and preparing chlorophyllin copper and sodium salts |
| WO2009100605A1 (en) * | 2008-01-30 | 2009-08-20 | Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd | A pharmaceutical composition for treating diabetes and preparation methods thereof |
| CN104095178A (en) * | 2013-04-09 | 2014-10-15 | 北京宏泰康达医药科技有限公司 | Composition used for assistant reduction of blood sugar |
| CN104415082A (en) * | 2013-09-11 | 2015-03-18 | 安阳天尊生物工程有限公司 | Trichosanthes kirilowii flesh extract and extraction method thereof as well as application of trichosanthes kirilowii flesh extract in treatment of diabetes mellitus |
-
1999
- 1999-06-14 CN CNB991091116A patent/CN1152692C/en not_active Expired - Lifetime
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100390176C (en) * | 2004-10-12 | 2008-05-28 | 大连理工大学 | Method for extracting chlorophyl from silkworm faeces by microwave pretreatment and preparing chlorophyllin copper and sodium salts |
| WO2009100605A1 (en) * | 2008-01-30 | 2009-08-20 | Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd | A pharmaceutical composition for treating diabetes and preparation methods thereof |
| GB2469220A (en) * | 2008-01-30 | 2010-10-06 | Zhangzhou Pien Tze Huang Pharm | A pharmaceutical composition for treating diabetes and preparation methods thereof |
| GB2469220B (en) * | 2008-01-30 | 2012-10-24 | Zhangzhou Pien Tze Huang Pharm | Pharmaceutical composition for treating diabetes and preparation method thereof |
| CN104095178A (en) * | 2013-04-09 | 2014-10-15 | 北京宏泰康达医药科技有限公司 | Composition used for assistant reduction of blood sugar |
| CN104415082A (en) * | 2013-09-11 | 2015-03-18 | 安阳天尊生物工程有限公司 | Trichosanthes kirilowii flesh extract and extraction method thereof as well as application of trichosanthes kirilowii flesh extract in treatment of diabetes mellitus |
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| Publication number | Publication date |
|---|---|
| CN1152692C (en) | 2004-06-09 |
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