Nghiên cứu và xây dựng quy trình ức chế hoạt tính của enzyme α glucosidase từ lá cóc đỏ (lumnitzera litterea)

- - BÁO CÁO Tên đề tài: Ê Ứ Ứ -GLUCOSIDASE Ừ (LUMNITZERA LITTOREA) S UY :T GVHD: SVTH: MSSV: 1153010176 -2015 Tp hí Minh, ngày 05 tháng 05 năm 2015 Trong su t th i gi n h Th nh ph v truy n hí t p t i kho inh qu t p tr n gi ng h ng n nghi n u t n t nh gi p ho n th nh t t kh n ng i nh h h nh ng ih y ng vi guy n Th o ho m su t s u n y Thủy v Th s d nh r t nhi u th i gi n v t m huy t h ớng h o v t o m i i u ki n thu n l i ho m lu n t t nghi p m h n th nh g i l i o em nh ng m v kho h T i xin nghi m Sinh s ng m n qu Th y ng nh n Th i – Tr áu l m n n t ng v ng h ng m xin g i l i i t n s u s guy n Th gh Sinh m xin h n th nh t ho m nh ng ki n th nh ng tháng năm h ng m n m n n Th u ủ nghi n nt t n ủng h t i v m t tinh th n s guy n inh u v t o ho m ni m ng l m vi o ng m ph ng thí m ho n th nh t t kh lu n t t nghi p V tr n h t on xin g i l i H iv on th hăm s y hi n t t huy n Em n n n l m n t t n áy l ng m nh i mt kh tinh th n v ng h n v ồng th i l nh ng l lu n ủ m nh m n Tp hí inh tháng 05 năm 2015 g i th guy n Th hi n im i Ẽ ii Ầ Ổ Ổ Ề LUMNITZERA 1.1.1 Phân lo i 1.1.2 i m h nh thái 1.1.3 h n 1.1.4 iá tr kinh t 1.1.5 T nh h nh nghi n Ổ u n v ngo i n Ề 1.2.1 guy n t tá h hi t 1.2.2 ỹ thu t hi t lỏng - lỏng 1.2.3 ỹ thu t hi t r n - lỏng 1.2.4 ỹ thu t hi t ph r n 11 1.2.5 y ut nh h ng 1.3 1.3.1 n tr nh tá h hi t nguy n li u 12 15 hái ni m 15 1.3.2 h n lo i 15 1.3.3 h 1.4 Ổ ng pháp i u tr 16 Ề α-GLUCOSIDASE 17 1.4.1 Enzyme α-glucosidase 17 h t 1.4.2 h nzym 18 1.5.3 T nh h nh nghi n u v kh h nzym α-glucosidase 21 Ầ Ê Ứ 23 V t li u 23 2.1 it 2.1.1 ng nghi n u 23 2.1.2 i m nghi n u 23 2.1.3 h t v thi t 24 2.2 Quy tr nh i u h o tổng 25 2.3 Quy tr nh i u h o ph n o n 26 ghi n 2.4 us nh h ng th i gi n nv nt ph n ng ủ enzyme -glucosidase 29 ghi n 2.5 us nh h ng ủ nồng h t nv nt ph n ng ủ enzyme -glucosidase 31 ghi n 2.6 u kh o hi t ỏ 33 2.7 So sánh kh v 2.8 Xá y nzym -glu osi s t h ho t tính ủ h nzym -glu osi s t o hi t y ỏ Tr ng 36 nh ki u h ho t tính nzym -glu osi s Ầ ủ o hi t 38 40 3.1 t qu i u h o tổng t 3.2 t qu i u h o ph n o n t 3.3 t qu nghi n us nh h y ỏ 40 o tổng 40 ng ủ th i gi n nv nt ph n ng enzyme -glucosidase 40 3.4 t qu nghi n us nh h ng ủ nồng h t nv nt ph n ng enzyme -glucosidase 43 3.5 t qu nghi n ủ 3.6 u kh th ho t tính o hi t t y So sánh kh y ỏ với ỏ 44 h y nzym -glucosidase h nzym -glu osi s ủ Xá nh ki u h ho t tính nzym -glu osi s 3.8 Xá nh ki u h ho t tính nzym -glu osi s Ầ o hi t Tr ng 49 3.7 o hi t thyl o hi t n-hexane 52 ủ t t 55 Ề 59 4.1 t lu n 59 4.2 i n ngh 64 65 iii B ng 1.1: Ưu v nh i m củ ph ng pháp hi t Soxhlet 10 nh h B ng 2.1: B trí thí nghi m nghiên c u s ng p-nitrophenyl-α-D-Glucosidase .32 B ng 2.2: B trí thí nghi m nghiên c u kh ủ ng 2.4: nzym α-glucosidase o hi t 33 B ng 2.3: B trí thí nghi m so sánh kh t h o hi t y ỏv trí thí nghi m xá h y nh ki u nzym α-glucosidase Tr ng 37 h ủ o hi t l n enzyme α-glucosidase 39 B ng 3.1: S nh h ng th i gi n n v n t c ph n ng .41 B ng 3.2: t qu nghi n nh h B ng 3.3: t qu kh c ch enzyme -glucosidase củ us ng p-nitrophenyl-α-D-glucosidase 43 o hi t 45 B ng 3.4: iá tr IC50 th hi n kh c ch enzyme -glucosidase cao chi t t y ỏ .47 B ng 3.5: iá tr IC50 cao chi t củ ng 3.6: t qu m t y qu ng th o s th y ổi nồng 4-Nitrophenyl α- - lu opyr nos ng 3.7: ng x l s li u m u kh ng ng 3.8: t qu m t ng x l s li u m u kh ng h t v nồng o hi t n-hexane 53 h v m u o hi t n-hexane 54 qu ng th o s th y ổi nồng 4-Nitrophenyl α- - lu opyr nos ng 3.9: ỏ Cóc Tr ng 50 v nồng h v m u h t o hi t ethyl acetate.56 o hi t thyl t t .57 i DANH M C HÌNH VẼ nh 1.1 : Lumnitzera littorea (Jack) Voigt nh 1.2: Lumnitzera racemosa Willd nh 1.3: o ủ L littorea nh 1.4: o ủ L racemosa nh 1.5: o ủ h i lo i .4 nh 1.6: t ủ h i lo i nh 1.7: th h nh 1.8: th h phi nh 1.9: th h kh ng nh 1.10: th nh 2.1: nh tr nh 19 nh tr nh 19 nh tr nh 20 h h n h p .21 y ỏ (Lumnitzera littorea) tr v s u ph i 23 nh 2.2: Quy tr nh i u h o tổng nh 2.3: Quy tr nh i u h o ph n o n nh 2.4: S nghiên c u s nh h y ỏ .25 y ng th i gi n ỏ 27 n v n t c ph n ng enzyme -glucosidase .30 nh 2.5: S nghi n u kh h nh 2.6: S huy n h h t n n nzym α-glu osi s th th hi n s nh h nh 3.1: nzym α-glu osi s t ng th i gi n o hi t .34 h 35 n v n t c ph n ng enzyme -glucosidase .42 nh 3.2: th th hi n kh c ch enzyme -glucosidase cao chi t t y ỏ 46 ii nh 3.3: th th hi n giá tr IC50 cao chi t t y ỏ Cóc Tr ng so với acarbose c ch enzyme -glucosidase .51 nh 3.4: th i u i n V th o S với nồng nh u ủ o hi t n-hexane 55 nh 3.5: th i u i n V th o S với nồng o hi t thyl nh u ủ t t .58 iii Ặ nh tháo i n h ng nh hi … ng l m t ăn U T T Ấ Ề nh nguy hi m v nh tim m h t i i n m h máu n o m l tr th nh nguy n nh n g y t vong th t Tr n th giới 10 gi y ồng hồ l i nh tháo tháo ng ng V v y vi ng m t ng nghi n nh kho h Trong h ớng i u tr v h ho t ng nzym α-glucosidas nh thu thu igu ni s tá Sulphonylur s n ỏ th m nhi u l u h t s h n ho vi tr n th giới n n nguồn th i t t m ki m v s nh hép h ng hi u i t v o th UY is ụng tá i u tr i u tr v nhi u tá n ụng phụ v giá kim lo i; y v ng phát ụng phụ n to n ho ng nh tháo nhi u is ng ph i nghi n ng nh ụng y ỏ v t ồi ng tr nh nghi n Vi t m nằm v ng khí h u y ỏ t nhi n l m th l ih y m t v th nh t h ho t tính o v phong ph T th i x x nh h y nh ng kỹ thu t hi n Ư uv l p h p h t t nhi n h i ng nghi p phát tri n y ỏ t nhi n v n T h h ng lo i thu h truy n mi ng v ghi nh ng nghi n i ng h ăn v thu k t th nh kinh nghi m qu nhi u th h n i ti p nh u ho n y m y h l nh v nh kho h ụng hi n n y v n nguy n li u nh u Trong nhi t ới gi m t ằng h ng í hh n h ho t tính enzyme α-glucosidas t ng lo i h ớng i u tr h ho t tính enzyme α-glucosidas t nhi u nguồn nh u nh kho h sinh h nh ụng phụ g y r i lo n ti u h ;… N n nhằm mụ ụng v ng pháp i u tr ụng phụ uồn n n ti u h y tá ng phát tri n i h t v nh ng i n h ng ủ n to n ho ng ng o n qu n t m nh tháo n gi n v suy th n n u ph h ho t tính enzyme α-glucosidas hi n h ng gi tăng nh nh với uv nguồn th n ng y h m t phẩm ồi nh ủ n U T T R ng ng p m n l m t h sinh thái huy n ti p gi li n tr ng nh kho h v ng qu n t m ng phong ph nguồn s ng v n i n nhi t ới v i t ho t tính sinh h g (Sonneratia caseolaris) n Vi t n hu v t th n g thu y ỏ hi Lumnitzera h h sinh thái ủ r ng ng p m n ng t n sá h ỏ m năm 1996 Tr n s t i “Nghiên cứu enzyme -glucosidase t hí tr l ng tá nghi n r ng ng p m n tr nhi u lo i l i n (Clerodendrum inerme), Ô rô (Acanthus (Lumnitzera littorea) l m t lo i th (Combretaceae) ph n nhi u m i n (Avicennia marina) tr ng (Lumnitzera racemosa), v.v Trong ilicifolius), t nl n it ph ps v t qu hi m nh ng i n v n nhi t ới go i r r ng ng p m n ủ nhi u hủng th li u m i tr ng th nh o ụ it í h ủ Cóc Đỏ (Lumnitzera ittorea)” u kh ng l m nh h ng nghi n ho t tính sinh h nguy n li u l nh v UY T Ư xu t Tiêu ng tới h sinh thái th u ho t i ot ho vi y li u v th nghi n v t ủ h n l nh ng t i g p ph n l m tăng giá tr v m t kho h nh kinh t cho loài này, t o ti n nhi n ng qu tr nh ức ch ho t t nh u ỏ g p ph n l m ng l p h p h tt ng h nguồn phẩm U  T T Qu tr nh điều ch cao ph n đo n: o tổng (200 g) n-hexane Chi t ki t h Cô quay o Ethyl acetate Cao nhexane (50.33 g) nl il n1 Chi t ki t h Cao ethyl o Ethyl acetate : ethanol (8:2) acetate (57.44 g) Cô quay nl il n2 Chi t ki t Cao ethyl h acetate : ethanol (8:2) (11.12 g) Cô quay h Cao ethanol (66.43 g) nh 4.2: Quy tr nh i u h UY T Ư o ph n o n y ỏ 62 U  T T Quy trình thử ho t tính enzym α-glucosidase: ml enzym α-glucosidase (0.5mg/ml) ml o hi t (10-100 μg ml) u Ủ 20 ph t nhi t 1ml p-nitrophenylα-D-glucosidase (5mM) ph ng u Ủ 20 ph t o nhi t qu ng ph ng 405 nm Tính toán iá tr Hình 4.3: Quy tr nh th ho t tính UY T Ư 50 h nzym α-glucosidase 63 U 4.2 T T Kiến nghị o th i gi n th hi n nghi m th o t i nh n h nhằm ho n thi n n n nghi n u ng tr nh nghi n ut i nh n h m ts ngh s u:  Nghiên c u th nghi m kh c ch enzyme α-glucosidase cao chi t tr n  it Nghiên c u quy trình cô l p h p ch t t nhiên có kh c ch enzyme α- glucosidase t  ng chu t thỏ ph n o n cao chi t S dụng cao chi t thu acetate : th nol (8:2) v c củ th nol c bi t cao chi t n-hexane ph n o n n-hexane, ethyl acetate, ethyl kh o sát ho t tính sinh h c v kh c ch enzyme α-amylase, tyrosinase enzyme khác UY T Ư 64 U T T TÀI LI U THAM KH Apostolidis E., Kwon Y.I and Shetty K (2007), Inhibitory potential of herb, fruit, and fungal-enriched cheese against key enzymes linked to type diabetes and hypertension, Innovative Food Science and Emerging Technologies, 8, 46-54 Australian Diabetes Council (2012), Diabetes: what you need to know, Copyright Australian Diabetes Council, 6-90 Bandaranayake W M (2002), Bioactivities, bioactive compounds and chemical constituents of mangrove plants, Wetlands Ecology and Management, 10, 421-452 c Tu n, Tr n Th Ki u nh át Văn Th nh guy n nh Quí (2002), Khu d tr sinh quy n r ng ng p m n C n Gi , Nhà xu t b n Nông nghi p, trang 32-80 FAO Regional Office for Asia and the Pacific (2007), Trees and shrubs of the Maldives, first edition, 195 Ganiyu O., Ayodele Jacob Akinyemi and Adedayo Oluwaseun Ademiluyi (2012), Antioxidant properties and inhibitory effect of athanolic extract of Struchium sparganophora (ewuro odo) leaf on α-amylase and α-glucosidase activities, Afr J Tradit Comphement Altern Med, (3), 342-349 Ganiyu O., Ayodele Jacob Akinyemi and Adedayo Oluwaseun Ademiluyi and Fatai Olumide Bello (2013), Inhibition of α-amylase and α-glucosidase activities by ethanolic extract of Amaranthus cruentus leaf as affected by blanching, (17), 1026-1032 Jun W., Qiang X., Jing X., Min-Yi L., Jian-Yu P and Mei-hua Y (2008), Natural products from true mangrove flora: source, chemistry and bioactivities, Natural product reports, 955-981 Lê Công Khanh (1986), R ng ng p m n r ng nhi t ới t chua phèn, Nhà xu t b n Thành ph Hồ Chí Minh, trang 40-80 UY T Ư 65 U T T 10 Lin T C., Hsu F L and Cheng J T (1993), Journal of Natural Products, 56, 629 11 Lisette D., Solimabi W & Prabha D (2010), Antibacterial phenolics from the mangrove Lumnitzera racemosa, Indian Journal of Marine Sciences, 39 (2), 294-298 lo 12 inhi itors: om s S h mi l stru tur rv lho (2006) “α and β-glucosidase n iologi l tivity” T tr h ron 62, 10277-10302 13 Miaomiao G., Renchao Z., Yelin H., Jianhua O., Suhua S (2011), Molecular confirmation of natural hybridization between Lumnitzera racemosa and Lumnitzera littorea, Aquatic Botany, 95, 59-64 14 Nguy n ng (2012), Công ngh enzyme, Nhà xu t b n i H c Qu c Gia TP Hồ Chí Minh, trang 34-60 15 Nguy n Kim Phi Phụng (2007), h xu t b n ng pháp l p h p ch t h u , Nhà i H c Qu c Gia TP.HCM, trang 20-53 16 Ph m Hoàng H (1999), Cây cỏ Vi t Nam, quy n 2, Nhà xu t b n trẻ, trang 110 17 h m Th Tr n nzym v h u ng ụng h n Tu n h xu t gh (2009) ng ngh sinh h n iáo ụ Vi t m trang 56-80 t p 3: 18 Premnathan M (1992), A survey of some Indian marine plants for antiviral activity, Botanica Marina, 35, 321-324 19 Priyarani M., Padmakumari K.P., Sankarikutty B., Lijo cherian O., Nisha V.M and Raghu K.G (2011), International Journal of Food Sciences and Nutrition, 62 (2): 106-110 20 Shabbudin S., Muhammad T., Deny S., Haitham Q., Nurul Afifah B Abdul R (2011), Antimicrobial activity of mangrove plant (Lumnitzera littorea), Asian Pacific Journal of Tropical Medicine, 523-525 21 Sundaram R., Murugesan G (2011), Hepatoprotective and antioxidant activity of a mangrove plant Lumnitzera racemosa, Asian Pacific Journal of Tropical Biomedicine, 348-352 UY T Ư 66 U T T 22 Tr n Mỹ Linh, Lê Quỳnh Liên, Ninh Kh c B n (2013), Ho t tính c ch n m vi khuẩn gây b nh ba loài th c v t ng p m n Aegiceras corniculatum, Avicennia marina Lumnitzera racemosa t i v n qu c gia Xuân Thủy, H i ngh khoa h c toàn qu c v sinh thái tài nguyên sinh v t l n th 5, trang 1127-1131 23 Verma N., Singh A P., Amresh G., Sahu P K (2010), “Different approaches for treatment of type diabetes mellitus with special reference to traditional medicines: a revi w” The Pharma Research, 3, 27-50 24 Selvam V (2007), Trees and shrubs of the Maldives, FAO Regional Office for Asia and the Pacific Printed by Thammada Press Co., Ltd., Bangkok, 195 25 Chapman V.J (1975), Mangrove vegetation, Auckland University, New Zealand, 447 UY T Ư 67 ảng ết ng iên cứu ản ưởng p-nitrophenyl-α-D- glucosidase ANOVA Table for Col_2 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 0.00162228 0.000324456 1168.04 0.0000 Within groups 0.00000333333 12 2.77778E-7 Total (Corr.) 0.00162561 17 Table of Means for Col_2 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 0.0193333 0.00030429 0.0188645 0.0198021 B 0.0276667 0.00030429 0.0271979 0.0281355 C 0.0353333 0.00030429 0.0348645 0.0358021 D 0.0433333 0.00030429 0.0428645 0.0438021 E 0.045 0.00030429 0.0445312 0.0454688 F 0.0436667 0.00030429 0.0431979 0.0441355 -Total 18 0.0357222 Multiple Range Tests for Col_2 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -A 0.0193333 X B 0.0276667 X C 0.0353333 X D 0.0433333 X F 0.0436667 X E 0.045 X -Contrast Difference UY T Ư iii ảng 3.5 ết k ả ức c ế enzyme -glucosi ase cao c iết Cao n-hexane: ANOVA Table for Col_2 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 3325.99 1108.6644346576.33 0.0000 Within groups 0.0002 0.000025 Total (Corr.) 3325.99 11 Table of Means for Col_2 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 27.2733 0.00288675 27.2686 27.278 B 42.4233 0.00288675 42.4186 42.428 C 54.55 0.00288675 54.5453 54.5547 D 72.7233 0.00288675 72.7186 72.728 -Total 12 49.2425 Multiple Range Tests for Col_2 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -A 27.2733 X B 42.4233 X C 54.55 X D 72.7233 X -Contrast Difference Ethyl acetate: ANOVA Table for Col_3 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 2429.25 809.74924292474.25 0.0000 Within groups 0.000266667 0.0000333333 Total (Corr.) 2429.25 11 UY T Ư iv Table of Means for Col_3 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 19.2333 0.00333333 19.2279 19.2388 B 30.7567 0.00333333 30.7512 30.7621 C 42.3067 0.00333333 42.3012 42.3121 D 57.6867 0.00333333 57.6812 57.6921 -Total 12 37.4958 Multiple Range Tests for Col_3 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -A 19.2333 X B 30.7567 X C 42.3067 X D 57.6867 X -Contrast Difference Cao ethyl acetate : ethanol (8:2): ANOVA Table for Col_2 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 3898.42 1299.4751978897.22 0.0000 Within groups 0.0002 0.000025 Total (Corr.) 3898.42 11 Table of Means for Col_2 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 8.70333 0.00288675 8.69863 8.70804 B 21.73 0.00288675 21.7253 21.7347 C 39.1267 0.00288675 39.122 39.1314 D 56.5233 0.00288675 56.5186 56.528 -Total 12 31.5208 UY T Ư v Multiple Range Tests for Col_2 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -A 8.70333 X B 21.73 X C 39.1267 X D 56.5233 X -Contrast Difference Cao ethanol ANOVA Table for Col_3 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 1378.1 459.36727561992.67 0.0000 Within groups 0.000133333 0.0000166667 Total (Corr.) 1378.1 11 Table of Means for Col_3 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 16.6733 0.00235702 16.6695 16.6772 B 27.28 0.00235702 27.2762 27.2838 C 38.89 0.00235702 38.8862 38.8938 D 44.4433 0.00235702 44.4395 44.4472 -Total 12 31.8217 Multiple Range Tests for Col_3 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -A 16.6733 X B 27.28 X C 38.89 X D 44.4433 X -Contrast Difference UY T Ư vi Bảng 3.6: Gía trị IC50 thể khả ức chế enzyme -glucosidase cao chiết từ óc ỏ ANOVA Table for Col_5 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 24089.2 6022.31********* 0.0000 Within groups 0.000333333 10 0.0000333333 Total (Corr.) 24089.2 14 Table of Means for Col_5 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 34.7733 0.00333333 34.7681 34.7786 B 71.1633 0.00333333 71.1581 71.1686 C 80.7667 0.00333333 80.7614 80.7719 D 143.283 0.00333333 143.278 143.289 E 33.4333 0.00333333 33.4281 33.4386 -Total 15 72.684 Multiple Range Tests for Col_5 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -E 33.4333 X A 34.7733 X B 71.1633 X C 80.7667 X D 143.283 X -Contrast Difference UY T Ư vii ảng 7: Gi trị 50 cao c iết p ân đoạn n-hexane, ethyl acetate, et yl acetate et anol (8 ), et anol óc ỏ óc rắng Cao n-hexane: ANOVA Table for Col_2 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 46498.9 23249.5*********** 0.0000 Within groups 0.0002 0.0000333333 Total (Corr.) 46498.9 Table of Means for Col_2 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 34.7667 0.00333333 34.7609 34.7724 B 186.573 0.00333333 186.568 186.579 C 33.4333 0.00333333 33.4276 33.4391 -Total 84.9244 Multiple Range Tests for Col_2 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -C 33.4333 X A 34.7667 X B 186.573 X -Contrast Difference Cao ethyl acetate: ANOVA Table for Col_3 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 11252.0 5626.01*********** 0.0000 Within groups 0.0002 0.0000333333 Total (Corr.) 11252.0 UY T Ư viii Table of Means for Col_3 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 71.1567 0.00333333 71.1509 71.1624 B 119.813 0.00333333 119.808 119.819 C 33.4333 0.00333333 33.4276 33.4391 -Total 74.8011 Multiple Range Tests for Col_3 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -C 33.4333 X A 71.1567 X B 119.813 X -Contrast Difference Cao ethyl acetate : ethanol (8:2): ANOVA Table for Col_4 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 14014.7 7007.37*********** 0.0000 Within groups 0.0002 0.0000333333 Total (Corr.) 14014.7 Table of Means for Col_4 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 80.7733 0.00333333 80.7676 80.7791 B 130.087 0.00333333 130.081 130.092 C 33.4333 0.00333333 33.4276 33.4391 -Total 81.4311 UY T Ư ix Multiple Range Tests for Col_4 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -C 33.4333 X A 80.7733 X B 130.087 X -Contrast Difference Cao ethanol: ANOVA Table for Col_5 by Col_1 Analysis of Variance Source Sum of Squares Df Mean Square F-Ratio P-Value Between groups 56895.6 28447.8*********** 0.0000 Within groups 0.0002 0.0000333333 Total (Corr.) 56895.6 Table of Means for Col_5 by Col_1 with 95.0 percent LSD intervals -Stnd error Col_1 Count Mean (pooled s) Lower limit Upper limit -A 143.283 0.00333333 143.278 143.289 B 227.633 0.00333333 227.628 227.639 C 33.4333 0.00333333 33.4276 33.4391 -Total 134.783 Multiple Range Tests for Col_5 by Col_1 -Method: 95.0 percent Duncan Col_1 Count Mean Homogeneous Groups -C 33.4333 X A 143.283 X B 227.633 X -Contrast Difference UY T Ư x g iên cứu k ả t oạt t n ức c ế enzyme -glucosi ase cao c iết từ óc ỏ óc rắng ng: h ng tr nh n tính hồi quy nghi n enzyme -glucosidase ủ h u kh th ho t tính h o hi t ph n o n ng tr nh n tính hồi quy ghi m th y Cao n-hexane Cao ethyl acetate Cao ethyl acetate : ethanol (8:2) Cao ethanol Acarbose ( UY T i h ng) Ư ỏ y Tr ng y = 44.695x - 18.885 y = 28.7x - 15.173 R² = 0.9898 R² = 0.9948 y = 38.117x - 20.602 y = 31.375x - 15.213 R² = 0.9824 R² = 0.9839 y = 48.248x - 42.021 y = 32.958x - 19.681 R² = 0.988 R² = 0.9892 y = 28.768x - 12.029 y = 28.013x - 16.033 R² = 0.9866 R² = 0.9868 y = 23.188x + 14.657 R² = 0.9948 xi ... 1.1.5 Tình hình nghiên cứu nước nước 1.1.5.1 C Cóc Đỏ (Lumnitzera littorea)  Trong n ớc: ho h h nn y h t m th y v ho t tính sinh h ủ ng tr nh nghi n y u kho h n o v th nh ph n ỏ (Lumnitzera littorea)... insulin… nh ph ng lo i ph n ti u i thi n h ng pháp 1.4 TỔNG QUAN VỀ ENZYME α-GLUCOSIDASE 1.4.1 Enzyme α-glucosidase[ 14] Enzyme α-glucosidase với nh ng tên khác nh maltase, glucoinvertase, glucosidoinvertase,... với u h t h enzyme, nh ng nzym làm tăng ho t tính enzyme mà chúng liên k t S liên k t ủ m t h t ho t h có th ngăn h n m t h t i vào tâm ng ủ enzyme gây tr ng i cho ph n ng xúc tác ủ enzyme h t