CN105842380B - A kind of method for screening α-amylase inhibitor - Google Patents
A kind of method for screening α-amylase inhibitor Download PDFInfo
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- CN105842380B CN105842380B CN201510014534.4A CN201510014534A CN105842380B CN 105842380 B CN105842380 B CN 105842380B CN 201510014534 A CN201510014534 A CN 201510014534A CN 105842380 B CN105842380 B CN 105842380B
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Abstract
The invention discloses a kind of method for screening α-amylase inhibitor, methods described includes：Enzymatic reaction buffer solution, substrate solution, alpha amylase solution, chromogenic reagent solution and need testing solution are prepared, it is standby；By need testing solution point sample on lamellae, flinging to after solvent or being impregnated in substrate solution after being dried using suitable solvent expansion, then take out, volatilize solvent, then be impregnated in alpha amylase solution, enzymatic reaction 10~60 minutes is carried out at 5~50 DEG C；After the completion of reaction, lamellae is taken out, volatilizes to be impregnated in after solvent in chromogenic reagent solution and carries out chromogenic reaction；Inspected under visible light：Blue spot under the white or yellow or brown background of appearance is alpha amylase inhibitor.The invention provides a kind of method that can quickly and easily screen α-amylase inhibitor, a desirable route is provided to filter out the research with alpha amylase inhibitory activity composition from natural materials.
Description
Technical field
The present invention is to be related to a kind of method for screening α-amylase inhibitor, specifically, is to be related to one kind to utilize thin layer color
The method of spectrum-bioautography technology screening α-amylase inhibitor.
Background technology
Diabetes be it is a kind of because internal insulin is absolute or relative hyposecretion caused by incretion metabolism disease, with height
The feature of fatal rate, high disability rate and high payment for medical care, is that the third-largest serious threat mankind are good for after tumour, cardio cerebrovascular affection
The Chronic Non-Communicable Diseases of health.Clinic antidiabetic drug has sulfonylureas, biguanides, Insulin sensitizing drugs, non-sulfonylureas pancreas islet at present
Element promotees to secrete medicine, glucosidase inhibitor etc., wherein, α-glucosidase inhibitor is by Reverse transcriptase small intestine epithelium suede
The effect of glycosidase on trichilemma reduces the degraded of carbohydrate, delays the digestion and absorption of carbohydrate, so as to be effectively reduced glycosuria
Patient's postprandial blood sugar concentration.
Pancreas α-amylase is another key enzyme in digestive system, belongs to α-glycosidase category.Pancreas α-amylase is to form sediment
The first step hydrolase of powder, by Starch Hydrolysis into small molecule oligosaccharides, including maltose, maltotriose and many α-(l -6) and α -
(l -4) oligosaccharides.These oligosaccharides are further degraded to glucose in the presence of α-glucuroide, are absorbed into blood circulation.Cause
This, pancreas α-amylase can cause the fast degradation to dietary starch, so as to raise postprandial blood sugar (PPHG).
In the 1970s, people come to realise, some inhibitor can suppress or part suppresses small-intestinal disaccharidases and pancreas
The activity of α-amylase, these inhibitor can be for oral medication non-insulin-depending type (diabetes B) diabetes.Cause
This, α-amylase inhibitor is significant in terms of diabetes B is treated.At present, the clinical α-amylase mainly applied
There is a series of side effects in inhibitor such as acarbose etc., for example：Flatulence, stomachache, diarrhoea etc..By contrast, natural goods
α-the amylase inhibitor in matter source because its action temperature and, the advantage such as small, wide material sources of toxic side effect highlight, therefore, ground
The extensive concern for the person of studying carefully.
And internal, the external kinds of experiments screening technique that prior art is used, the purity and consumption to test compound is equal
Have higher requirements, there is the defects such as screening difficulty is big, cost is high.Especially, the active ingredient of natural materials is generally not clear and definite enough,
And active effect is produced by the synergy of multicomponent, Mutiple Targets mostly, using the screening technique of prior art to from day
When α-amylase inhibitor of right material is screened, there can be bigger difficulty, not only time and effort consuming, and it is complicated poorly efficient.Cause
This, studies a kind of method that can quickly screen α-amylase inhibitor, to screening α-starch enzyme level from natural materials
Agent will be significant.
The content of the invention
In view of the above-mentioned problems existing in the prior art, it is an object of the invention to provide a kind of screening α-amylase inhibitor
Method, using TLC-bioautography technology, screens α-starch with realizing quick, low cost, high sensitivity and specificity
Enzyme inhibitor.
For achieving the above object, the technical solution adopted by the present invention is as follows：
A kind of method for screening α-amylase inhibitor, comprises the following steps：
A) enzymatic reaction buffer solution, substrate solution, alpha-amylase solution, chromogenic reagent solution and need testing solution are prepared, it is standby
With；Described enzymatic reaction buffer solution is from the Tri(Hydroxymethyl) Amino Methane Hydrochloride buffer solution that pH value is 6.5~9.0；It is described
Substrate solution from pH value be 6.0~7.0, the phosphate buffer solution of the soluble starch containing 0.05wt%~1wt%；It is described
Alpha-amylase solution select active concentration >=6U/mL containing alpha-amylase enzymatic reaction buffer solution；Described developer solution
Liquid is from the iodine-potassium iodide aqueous solution that content of iodine is 0.1~20mg/mL；
B) by need testing solution point sample on lamellae, fling to it is standby after solvent or using suitable solvent expansion after take
Go out, dry it is standby；
C) lamellae after being handled through step b) is impregnated in substrate solution, then takes out, volatilizes solvent；
D) lamellae after being handled through step c) is impregnated in alpha-amylase solution, enzymatic reaction is carried out at 5~50 DEG C
10~60 minutes；After the completion of reaction, take out lamellae, volatilize solvent；
E) lamellae after being handled through step d) is impregnated in chromogenic reagent solution and carries out chromogenic reaction；
F) lamellae after being developed the color through step e) is inspected：White or yellow or the brown back of the body occurred under visible light
Blue spot under scape is alpha-amylase inhibitor.
Preferably, described enzymatic reaction buffer solution is by trishydroxymethylaminomethane and hydrochloric acid and calcium chloride shape
Into the molar concentration of trishydroxymethylaminomethane therein is 25~80mmol/L.
Preferably, described phosphate buffer solution is formed by sodium dihydrogen phosphate with disodium hydrogen phosphate, and institute's shape
Into phosphatic molar concentration be 15~25mmol/L.
Preferably, described chromogenic reagent solution is 3 in mass ratio by KI and iodine:1 content of iodine formed is 1
~5mg/mL the aqueous solution.
Preferably, described lamellae selects any one in silica gel plate, polyamide board, cellulose plate.
Compared with prior art, the present invention has the advantages that and conspicuousness progress：
1) simple to operate without complicated instrument and equipment, experiment expends low, is adapted to common laboratory operation, Er Qieling
Sensitivity and high-specificity are in conventional screening assays；
2) the selection result is visual in image, it is easy to identification and memory；
3) screening process is not limited using lamellae as medium by solvent and test sample concentration so that the choosing of given the test agent
Select scope more extensive；
In a word, it is from natural goods the invention provides a kind of method that can quickly and easily screen α-amylase inhibitor
The research with alpha-amylase inhibitory activity composition is filtered out in matter there is provided a desirable route, with conspicuousness application valency
Value.
Brief description of the drawings
Fig. 1 is that embodiment 1 is inspected acarbose suppression α-diastatic activity sexuality using the screening technique of the present invention
As a result.
Fig. 2 is the result of the appropriate concentration range of contained soluble starch in substrate solution used in 2 pairs of embodiment；
Wherein：Soluble-containing starch is shallow lake containing solubility in 0.05wt%, the substrate solution used in B plates in substrate solution used in A plates
Powder is 1wt%.
Fig. 3 is the result of the appropriate concentration range of contained iodine in chromogenic reagent solution used in 3 pairs of embodiment；Wherein：A plates
Content of iodine is 0.1mg/mL in chromogenic reagent solution used, and content of iodine is 20mg/mL in the chromogenic reagent solution used in B plates.
Fig. 4 is the checking of the suitable activity concentration range of contained alpha-amylase in alpha-amylase solution used in 4 pairs of embodiment
As a result；Wherein：The active concentration containing alpha-amylase is 6U/mL, the alphalise starch used in B plates in alpha-amylase solution used in A plates
The active concentration containing alpha-amylase is 120U/mL in enzyme solutions.
Fig. 5 is the result of the embodiment 5 to the Suitable ranges of enzymatic reaction；Wherein：Enzymatic reaction used in A plates
Temperature is 5 DEG C, and the enzymatic reaction temperature used in B plates is 50 DEG C.
Fig. 6 is the result of the embodiment 6 to the suitable time scope of enzymatic reaction；Wherein：Enzymatic reaction used in A plates
Time is 10 minutes, and the time of enzymatic reacting used in B plates is 60 minutes.
Fig. 7 is that embodiment 7 inspects Comparative result to perilla seed, the fruit of glossy privet and Baical Skullcap root P.E under the conditions of difference is inspected；
Wherein：A figures are the results of inspecting under 254nm wavelength lights photograph, and B figures are to inspect result under 366nm wavelength lights photograph, and C figures are
Under visible light inspect result.
Fig. 8 is that embodiment 8 uses the screening technique of the present invention to Rosiglitazone, Miglitol, oleanolic acid and reseda
The selection result of the element to the rejection ability of α-amylase；Wherein：A is acarbose；B is Rosiglitazone；C is Miglitol；D is
Oleanolic acid；E is cyanidenon.
Embodiment
Technical scheme is further elaborated with reference to specific embodiment：
Embodiment 1
First, solution is prepared
The preparation of enzymatic reaction buffer solution：302.5mg trishydroxymethylaminomethanes and 330mg anhydrous calcium chlorides are dissolved in water
In, with salt acid for adjusting pH value to 7.5, then add water and be settled to 100mL, it is 7.5, mole of trishydroxymethylaminomethane to produce pH
Concentration is 25mmol/L enzymatic reaction buffer solution, is placed in 2~4 DEG C of refrigerators and saves backup.
The preparation of substrate solution：By 312mg NaH2PO4·2H2O and 716mg Na2HPO4·12H2O be dissolved in 100mL go from
In sub- water, regulation pH is 6.9, and it is the phosphate buffer solution that 6.9, phosphate concn is 20mmol/L to produce pH；Take 1g solvable
Property starch is dissolved in the above-mentioned phosphate buffer solutions of 100mL, after 95 DEG C of heating water baths 4 hours, is let cool, in 8000 revs/min
Lower centrifugation 10 minutes, gained supernatant is that pH value is the 6.9, substrate of the phosphate buffer solution of the soluble starch containing 1wt%
Solution, is placed in 2~4 DEG C of refrigerators and saves backup.
The preparation of alpha-amylase solution：Alpha-amylase is added in above-mentioned enzymatic reaction buffer solution, alpha-amylase is configured to
Active concentration be 6U/mL alpha-amylase solution.
The preparation of chromogenic reagent solution：By KI and iodine in mass ratio 3:1 is dissolved in deionized water, is configured to content of iodine and is
2mg/mL chromogenic reagent solution.
The preparation of need testing solution：5mg acarboses (Acarbose) powder is dissolved in 100mL methanol, is configured to dense
Spend the acarbose solution for 0.05mg/mL.
2nd, operation is inspected to the inhibitory activity of α-amylase
By above-mentioned need testing solution point sample on lamellae, parallel 3 points of point sample, point sample amount is 1 μ L；Volatilize methanol solvate
After be impregnated in above-mentioned substrate solution；Take out, volatilize solvent, then be impregnated in above-mentioned alpha-amylase solution, enzyme is carried out in 37 DEG C
Promote reaction 30 minutes；After the completion of reaction, take out lamellae, volatilize solvent, then be impregnated in above-mentioned chromogenic reagent solution and developed the color
Reaction；Inspected under visible light, inspect result as shown in Figure 1：All equal tables of need testing solution sampling point on lamellae
It is now the blue spot under yellow background, it is alpha-amylase inhibitor to illustrate acarbose, consistent with existing result of study, enters one
Step explanation the inventive method has feasibility.
Embodiment 2
The present embodiment the difference is that only with embodiment 1：Substrate solution used is respectively 6.9 from pH value, contained
The phosphate buffer solution and pH value of 0.05wt% soluble starches are that the phosphate-buffered of the 6.9, soluble starch containing 1wt% is molten
Liquid, remaining solution is identical with described in embodiment 1.
Need testing solution difference point sample (is abbreviated as in 2 lamellaes：A plates and B plates) on, 3 points of the parallel point sample of every plate, point
Sample amount is 1 μ L；Volatilize and be impregnated in after methanol solvate in above-mentioned substrate solution respectively；Take out, volatilize solvent, then be impregnated in α-shallow lake
In powder enzyme solutions, enzymatic reaction 30 minutes is carried out in 37 DEG C；After the completion of reaction, take out lamellae, volatilize solvent, then be impregnated in aobvious
Chromogenic reaction is carried out in toner solution；Inspected under visible light, inspect result as shown in Figure 2：Institute on A plates and B plates
There is need testing solution sampling point to show as the blue spot under yellow background, illustrate soluble shallow lake contained in substrate solution used
The concentration of powder is suitable in 0.05wt%~1wt%.
Embodiment 3
The present embodiment the difference is that only with embodiment 1：Content of iodine in chromogenic reagent solution used is respectively
0.1mg/mL and 20mg/mL, remaining solution is identical with described in embodiment 1.
Need testing solution difference point sample (is abbreviated as in 2 lamellaes：A plates and B plates) on, 3 points of the parallel point sample of every plate, point
Sample amount is 1 μ L；Volatilize after methanol solvate and be impregnated in substrate solution respectively；Take out, volatilize solvent, then be impregnated in alpha-amylase
In solution, enzymatic reaction 30 minutes is carried out in 37 DEG C；After the completion of reaction, lamellae taken out, volatilize solvent, then be impregnated in above-mentioned aobvious
Chromogenic reaction is carried out in toner solution；Inspected under visible light, inspect result as shown in Figure 3：Institute on A plates and B plates
There is need testing solution sampling point to occur in that the content of iodine in the blue spot under white or yellow or brown background, chromogenic reagent solution
Height be influence lamellae background color, with the increase of content of iodine, background colour by white change again to brown to yellow
Become, but do not influence to inspect result；Illustrate that the concentration of contained iodine in chromogenic reagent solution used is suitable in 0.1~20mg/mL.
Embodiment 4
The present embodiment the difference is that only with embodiment 1：Contained alpha-amylase in alpha-amylase solution used
Active concentration is respectively 6U/mL and 120U/mL, and remaining solution is identical with described in embodiment 1.
Need testing solution difference point sample (is abbreviated as in 2 lamellaes：A plates and B plates) on, 3 points of the parallel point sample of every plate, point
Sample amount is 1 μ L；Volatilize after methanol solvate and be impregnated in substrate solution respectively；Take out, volatilize solvent, then be impregnated in above-mentioned α-shallow lake
In powder enzyme solutions, enzymatic reaction 30 minutes is carried out in 37 DEG C；After the completion of reaction, take out lamellae, volatilize solvent, then be impregnated in aobvious
Chromogenic reaction is carried out in toner solution；Inspected under visible light, inspect result as shown in Figure 4：Institute on A plates and B plates
There is need testing solution sampling point to occur in that the blue spot under yellow background, illustrate contained α-shallow lake in alpha-amylase solution used
As long as active concentration >=6U/mL of powder enzyme is that can obtain preferably to inspect effect.
Embodiment 5
Solution used in the present embodiment is identical with described in embodiment 1.
Need testing solution difference point sample (is abbreviated as in 2 lamellaes：A plates and B plates) on, 3 points of the parallel point sample of every plate, point
Sample amount is 1 μ L；Volatilize after methanol solvate and be impregnated in substrate solution respectively；Take out, volatilize solvent, then be impregnated in alpha-amylase
In solution, enzymatic reaction 30 minutes (A plates) is carried out respectively at 5 DEG C and 50 DEG C carry out enzymatic reaction 30 minutes (B plates)；Reaction is completed
Afterwards, take out lamellae, volatilize solvent, then be impregnated in chromogenic reagent solution and carry out chromogenic reaction；Inspected, examined under visible light
It is as shown in Figure 5 depending on result：All need testing solution sampling points on A plates and B plates occur in that the blue spot under yellow background,
Illustrate that enzymatic reaction temperature, at 5~50 DEG C, is optimal with 37 DEG C.
Embodiment 6
Solution used in the present embodiment is identical with described in embodiment 1.
Need testing solution difference point sample (is abbreviated as in 2 lamellaes：A plates and B plates) on, 3 points of the parallel point sample of every plate, point
Sample amount is 1 μ L；Volatilize after methanol solvate and be impregnated in substrate solution respectively；Take out, volatilize solvent, then be impregnated in alpha-amylase
In solution, enzymatic reaction 10 minutes (A plates) is carried out respectively at 37 DEG C and 37 DEG C carry out enzymatic reaction 60 minutes (B plates)；React
Cheng Hou, takes out lamellae, volatilizes solvent, then be impregnated in chromogenic reagent solution and carry out chromogenic reaction；Inspected under visible light,
Inspect result as shown in Figure 6：All need testing solution sampling points on A plates and B plates occur in that the blue spot under yellow background
Point, illustrates that time of enzymatic reacting, at 10~60 minutes, was optimal with 30 minutes.
Embodiment 7
Solution used in the present embodiment the difference is that only with embodiment 1：Need testing solution used respectively by perilla seed,
The fruit of glossy privet and Baical Skullcap root P.E are made：Perilla seed, the fruit of glossy privet and each 0.5g of radix scutellariae medicinal materials fine powder are weighed, 5mL methanol is separately added into
In, ultrasonication is after 30 minutes, and centrifugation takes supernatant, produces perilla seed need testing solution, fruit of glossy privet need testing solution and Huang
A kind of reed mentioned in ancient books need testing solution.
Draw above-mentioned each 6 μ L of three kinds of need testing solutions respectively, point sample on same lamellae, fling to after methanol with chloroform-
(volume ratio is 9 to methanol-water-formic acid:1:0.1:0.05) deployed for solvent, take out lamellae and fully volatilize solvent
After be impregnated in substrate solution；Take out, volatilize solvent, then be impregnated in alpha-amylase solution, enzymatic reaction 30 is carried out in 37 DEG C
Minute；After the completion of reaction, take out lamellae, volatilize solvent, then be impregnated in chromogenic reagent solution and carry out chromogenic reaction；Exist respectively
254nm wavelength lights are inspected (A scheme) in such as Fig. 7, (the B figures in such as Fig. 7) are inspected under 366nm wavelength lights photograph according to lower
And (the C figures in such as Fig. 7) is inspected under visible light.From inspecting result under the conditions of different inspect：Under visible light
To inspect result consistent with existing result of study, i.e.,：Perilla seed, the fruit of glossy privet and Baical Skullcap root P.E have certain to α-amylase
Inhibitory activity, and the activity of perilla seed is more than the fruit of glossy privet, the activity of the fruit of glossy privet is more than radix scutellariae.
Embodiment 8
Using the enzymatic reaction buffer solution described in embodiment 1, substrate solution, alpha-amylase solution and chromogenic reagent solution and sieve
Choosing method, is examined to Rosiglitazone, Miglitol, oleanolic acid and cyanidenon to the inhibitory activity of α-amylase respectively
Depending on：
Reference substance solution：By acarbose (Acarbose, article No.：A8980-1G, purchased from Sigma) methanol is dissolved in, prepare
Into the solution that concentration is 0.05mg/mL；
Rosiglitazone need testing solution：By Rosiglitazone (Rosiglitazone, article No.：111M4728V, purchased from Sigma)
Methanol is dissolved in, the solution that concentration is 1mg/mL is configured to.
Miglitol need testing solution：By Miglitol (Miglitol, article No.：0446211-4, purchased from Cayman
Chemical) it is dissolved in deionized water solution, is configured to the solution that concentration is 1mg/mL.
Oleanolic acid need testing solution：By oleanolic acid (Oleanolic acid, article No.：07-2004, purchased from upper marine
Medicine Standardization Research center) methanol is dissolved in, it is configured to the solution that concentration is 1mg/mL.
Cyanidenon need testing solution：By cyanidenon (Luteolin, article No.：05-2008, purchased from Shanghai Chinese medicine standard
Change research center) methanol is dissolved in, it is configured to the solution that concentration is 1mg/mL.
By above-mentioned reference substance solution and need testing solution difference point sample on 5 lamellaes (successively labeled as A, B, C, D,
E), parallel 3 points of point sample, reference substance point sample amount is 1 μ L, and test sample point sample amount is 3 μ L, without expansion, volatilizes and is soaked after solvent
Stain is in substrate solution；Take out, volatilize solvent, then be impregnated in alpha-amylase solution, enzymatic reaction 30 minutes is carried out in 37 DEG C；
After the completion of reaction, take out lamellae, volatilize solvent, then be impregnated in chromogenic reagent solution and carry out chromogenic reaction；Enter under visible light
Row is inspected, and inspects result as shown in figure 8, in Fig. 8：A is acarbose；B is Rosiglitazone；C is Miglitol；D is olive
Acid；E is cyanidenon.As seen from Figure 8：Compared with acarbose, cyanidenon and Rosiglitazone show certain α-shallow lake
Powder enzyme inhibition activity, Rosiglitazone is relatively weak to the inhibitory activity of alpha-amylase, and Miglitol and oleanolic acid are then to α-shallow lake
Powder enzyme does not have inhibitory activity.
To sum up test visible：The inventive method need not be complicated instrument and equipment, can quickly, high flux screening go out to α-
Amylase has the composition of inhibitory activity, and not only the selection result is visual in image, readily discernible and memory, and sensitivity and exclusive
Property be higher than conventional screening assays, and do not limited by solvent and test sample concentration so that the range of choice of tested test sample
It is more extensive, it is adapted to common laboratory operation, a kind of convenient reality is provided with alpha-amylase inhibitory activity composition to filter out
Approach, with conspicuousness application value.
Finally be necessary described herein be：Above example is served only for making further detailed to technical scheme
Ground explanation, it is impossible to be interpreted as limiting the scope of the invention；Those skilled in the art is according to the above of the invention
Some the nonessential modifications and adaptations made belong to protection scope of the present invention.
Claims (4)
1. a kind of method for screening α-amylase inhibitor, it is characterised in that comprise the following steps：
A) enzymatic reaction buffer solution, substrate solution, alpha-amylase solution, chromogenic reagent solution and need testing solution are prepared, it is standby；Institute
The enzymatic reaction buffer solution stated, from the Tri(Hydroxymethyl) Amino Methane Hydrochloride buffer solution that pH value is 6.5~9.0, is by three hydroxyl first
Base aminomethane is formed with hydrochloric acid and calcium chloride, and the molar concentration of trishydroxymethylaminomethane therein is 25~80mmol/L；
Described substrate solution is 6.0~7.0, phosphate buffer solution of the soluble starch containing 0.05wt%~1wt% from pH value；
Described alpha-amylase solution selects the enzymatic reaction buffer solution of active concentration >=6U/mL containing alpha-amylase；Described colour developing
Agent solution is from the iodine-potassium iodide aqueous solution that content of iodine is 0.1~20mg/mL；
B) by need testing solution point sample on lamellae, fling to it is standby after solvent or using suitable solvent expansion after take out,
Dry standby；
C) lamellae after being handled through step b) is impregnated in substrate solution, then takes out, volatilizes solvent；
D) by through step c) handle after lamellae be impregnated in alpha-amylase solution, 5~50 DEG C carry out enzymatic reactions 10~
60 minutes；After the completion of reaction, take out lamellae, volatilize solvent；
E) lamellae after being handled through step d) is impregnated in chromogenic reagent solution and carries out chromogenic reaction；
F) lamellae after being developed the color through step e) is inspected：Under the white or yellow or brown background that occur under visible light
Blue spot be alpha-amylase inhibitor.
2. the method as described in claim 1, it is characterised in that：Described phosphate buffer solution is by sodium dihydrogen phosphate and phosphoric acid
Disodium hydrogen is formed, and the phosphatic molar concentration formed is 15~25mmol/L.
3. the method as described in claim 1, it is characterised in that：Described chromogenic reagent solution is in mass ratio with iodine by KI
3:The aqueous solution that 1 content of iodine formed is 1~5mg/mL.
4. the method as described in claim 1, it is characterised in that：Described lamellae selects silica gel plate, polyamide board, cellulose
Any one in plate.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102980969A (en) * | 2012-11-27 | 2013-03-20 | 上海中医药大学 | Method for screening lipase inhibitor |
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|---|---|---|---|---|
| CN102980969A (en) * | 2012-11-27 | 2013-03-20 | 上海中医药大学 | Method for screening lipase inhibitor |
Non-Patent Citations (3)
| Title |
|---|
| Digestion of legume starch granules by larvae of Zabrotes subfasciatus (Coleoptera: Bruchidae) and the induction of a–amylases in response to different diets;Carlos P. Silva et al;《Insect Biochemistry and Molecular Biology》;20011231;第31卷;41-50 * |
| Extracts of Maqui (Aristotelia chilensis) and Murta (Ugni molinae Turcz.): Sources of Antioxidant Compounds and r-Glucosidase/r-Amylase Inhibitors;Monica Rubilar et al;《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》;20110204;第59卷;1630-1637 * |
| 薄层色谱生物自显影法筛选新疆莫合烟天然乙酰胆碱酯酶抑制剂的活性成分;王健等;《中国药业》;20141020;第23卷(第20期);38-40 * |
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