• N g h i e n C I J U - Ky t h u a t
Do dOng
Xac dinh bang each them 75, 100, 125%
l u g n g e h u i n d6i ehieu vao mdu placebo (mdi
n i n g do t h y c hien 3 m l u ) . Trung binh l y lg phyc hoi (%) cac e h u i n doi chieu them vao dugc trinh bay trong bang 5.
Bang 5: Do dung cua phwang phap trong phan tich dinh lwang glucose va fructose, sorbitol va manitol trong miu thwc phim chwc nang
Mau Hcrp chat
Glucose Fructose Manitol Sorbitol
7594 ± RSD 100.59 + 0.81
99.59 ±0.18 100.35 + 0.53 98.91 ±0.71
Ty le phuc hoi (%) 100% ± RSD 99.29 ± 0.24 99.95 ± 0.39 100.22 ±0,39 100,34 ±0,57
125% t RSD 99,71 ± 0,21 99.18 ±0,51 100.71 ±0.40 99,91 ± 0,24
Tinh thong ke
T h u t-Test cho cac gia tri thu d u g c , k i t qua cho t h i y . eac gia tn Irung binh tren gidng nhau v§ mat thong ke, RSD< 2%. Vgy do diing cCia phuang phap 99,9 % la chap nhan d u g c .
Ket luan
Chung toi da xay dyng quy trinh d\nh tinh, dinh lugng d i n g thai d u d n g d a n la glucose va fructose, polyalcol d u d n g la manitol va sorbitol bang p h u a n g phap s i c ky long hieu nang cao vdi d i u do Rl. Quy trinh dap dng tdt cac yeu c l u cua phuang phap phan tieh. Do vgy, eo the ap dung quy trinh nay de dinh tinh, djnh lugng eae c h l pham cd chua glucose va fructose, manitol va sorbitol fren th| trudng.
Tai lieu tham khao 1. Bo Yti (2009). Duac dien Viet Nam IV.
2. Dethy J M., Callaert-Deveen B., Jansens M. (1984),
"Determination of sorbitol and galactitol in biological samples by HPLC", J. fijial Biochem., 142,119-124.
3. Food Chemicals Codex, 2008-2009, Edit 6, USP- United Book Press. Inc.
4 Vincente C. Mateos J. L (1991). "HPLC detemiinaton of sugar and pdyols in extracts of liciiens and sugarcane juice", J. Chromatogr., 553, 271-283
5. Yamachi S.. Nakai C , Nimura N. (1993),
"Highly sensitive detection of non-reducing carbohydrate by HPLC", J. Analyst., 118 (7), 769-771.
6. Kadoya S , Kahon F., Izutsu K. , Yonemochi E., Terada K , Chikako Y., Kawanishi T (2010), "Freeze- drying of proteins with glass-forming oligosacciiande- derived sugar alcohols", International Journal of Pharmaceutics. 389,107-113.
r -y
Phan lap mot so thanh phan hoa hoc tir cay rau ma ngo [Polygonum perfoliatum L.)
Nguyen Van Dau", D6 Thi Mai, Oinh Nhir Chien Khoa Hoa hoc. Trucmg BHKHTN-BHQGHN E-mail: [email protected]
Summary
Stearic acid (1), ^-sitosterol (2), daucosterol (3), 3,4-dihydro-5-hydroxy-7-methoxy-4-(4'- methoxyphenyl)coumarin (4) and quercetin (5) were isolated from the aerial parts of Polygonum perfoliatum L. by combination of various chromatography techniques. Their structures were identified by NMR. The occurrence of 3,4-Dihydro-5-hydroxy-7-methoxy-4-(4'-methoxyphenyl)coumarin (4) was detected for the first time in the Polygonym genus.
Keywords: Polygonum perfoliatum L., p-sitosterol, daucosterol, quercetin 3.4-dihydro-5-hydroxy-7- me-thoxy-4-(4'-methoxyphenyl)coumarln.
36 TAP CHi DUgfC HQC - 09/2013 (SO 449 NAM 53)
• Nghien CLFU - Ky thuat
Dat van de
Rau ma ngp (thom lom gai) c6 ten khoa hpc Id Polygonum perfoliatum L., thudc hg Rau ram (Polygonaceae^.
Theo Ddng y thom l6m gai eo vi d i n g , chua, tinh binh vao cac kinh can, than. Cd tac dung thanh nhiet giai dgc, hogt huyet t r u phong, Ihudng diing ehua ho ga. trT. vang da. sot ret.
kiet ly. lieu tign buot, khi hu,...'^'
Theo tra c d u ciia ehung tdi, cho den nay d Viet Nam chua ed cdng trinh nghien cdu vd thanh phdn hda hpc ciia cdy thudc nay. Bai bao nay thong bao ket qua phan lap mdt so thanh phan t u cay rau ma ngg va nhan dang chiing la acid stearic (1), p-sitosterol (2), daucosterol (3), 3.4-dihydro-5-hydroxy-7-methoxy-4-(4'-metoxyphenyl) -coumarin (4) va quercetin (5).
Nguyen lieu, thiet bj va phu'O'ng phap nghien ciJu
Nguyen lieu
Cay rau ma ngg {Polygonum perfoliatum L.) duge thu hai tgi Yen San, Ba Vi, Ha Ndi vao thang 10/2012 va duac PGS. Tran Van On, Tardng Oai hqc Dugc Ha Npi gidm dinh ten khoa hgc,
Hoa chat thiet bj
Ban mdng silica gel trang sSn DC-Alufolien 64 F254 (Merck. Dannstadt, CHLB Due); silica gel cho cdt sdc ki c d hgt 63-100 pm. Cae dung mdi (Trung Quoc) d u g c tinh che trudc khi tien h^nh sic ky.
Pho ' H - N M R , • ' ^ C - N M R d u g c do trdn may Bruker DRX500 ciia Vidn Hda hpc, Vien Khoa hpc va Cdng nghd Viet Nam.
Chiet nguyen li^u
Bdt khd phan tren mat dat ciia cay rau ma ngg (3 kg) dugc ngam kiet trong methanol 5 ldn, sau 66 Ipc lay djch chiet va cat logi bd dung mdi thu dugc cgn chiet methanol. Hda tan can methanol trong nudc cat va chiet lan l u g l vdi cac dung mdi n-hexan, dichlonDmethan, ethyl acetat.
n-butanol. Sau khi logi dung moi dudi ap s u i t giam thu d u g c cac cgn chief n-hexan (94,8 g, higu su^l 3,16%), dichloromelhan (8,13g, hieu su^t 0,27%), ethyl acetat (19,31 g, higu suat 0,64%) va n-butanol (9,52 g, higu suat 0,31%).
Phan lap cac chSt
C^c c^n chiet n-hexan, dichloromelhan va ethyl acetat d u g c phan tach tren cgt s i c ki
TAP C H i DUgfC HQC - 09/2013 (SO 449 N \ M 53)
silica gel vdi hdn hgp dung mdi-gradient, tang d^n do phan eye. Cac ehat phan lap d u g c tinh ehe tiep b i n g phuang phap k i t tinh tgi.
Ket qua va thao luan
Ket q u a phan lapBang phuang phap s§c ky cdl, t d can chief n- hexan (rua giai vdi hdn hgp n-hexan/elhyl acetat) thu d u a c c h i t (1) va (2); t u can chiet dichloromelhan (nia giai vdi hdn hgp n- hexan/aceton) nhgn dugc c h i t (3), va t u can chief ethyl acetat {ara giai vdi hdn hgp dichloromethan/aceton) thu dugc c h i t (4) va (5).
Xac d j n h cau true cua cac chat phan lap Acid stearic (1)
Hap c h i t 1 thu dugc dudi dang chdt rdn mau trdng (25 mg). Trong pho ^H-Nli^R (500 MHz, CDCb) xuat hien tin hieu ciia nhdm methyl d 6H 0,88 ppm (3H. t, J=6,75Hz), tin hidu ciia 14 nhdm methylen (5H 1,294-1,25 ppm), tin hieu hai nhdm methylen d 5H 2,26 ppm (2H, t, J = 7.75Hz) va d 5H 1,59 (2H, sextet, J=7.38Hz).
Pho "C-WMR (125MHz, CDCI3) chl ra tin hieu cua 18 carbon, trong dd cd mdt carbon ciia nhdm carboxylic d 5c = 194,562 ppm. mgt carbon ciia nhdm metylen vj tri a ddi vdi nhom carboxylic lai 5c = 99,042 ppm, va 16 carbon tai 5c 14,11 (C-18), 22,70 (C-17), 23,43 (C-16), 25,76 (C-15), 29,06 (C-14), 29,26 (C-13), 29,36 (C-12). 29,48 (C-11). 29,48 (C-10), 29,62 (C-9), 29,66 (C-8), 29.68 (C-7), 29,93(C-6). 38,43(C- 5), 43,82 (C-4). 57.23 (C-3)
Nhu vgy pho NMR eiia chat (1) phii hgp vdi pho ciia mgt acid no, mach dai, phan t u cd 18 carbon la acid steanc.
P-sitosterol (2)
Tinh the hinh kim mau trang (35 mg). Nhiet dp ndng chay 135-138°C. Chat (2) cd R, triing vdi eua p-sitosterol ehuan trong cung mdt didu kien s i c ki Idp mdng. Pho ^H-NMR (500 MHz, CDCI3) ciia chdt (2) cung khang dinh cau triic nay thdng qua cae pic dac trung ciia proton- carbinol d 5H 3,68 ( I H , m. H-3). proton melin trong vdng d 5H 5,33 ( I H , d, J=5,2 Hz, H-6), sau nhdm methyl d 5H 0.96 (3H, d, J=7,0 Hz, H-21), 0,84 (3H. t, J=7.0 Hz. H-26), 0,86 (3H, d, J=8,0 Hz, H-27), 0,88 ( 3H, d, J=7.5 Hz, H-29), 0,76 (3H, s, H-18), 1,03(3H, s, H-19).
37
• Nghien ctru - Ky thuat
p-sitosterol Daucosterol (3)
C h i t ran mau t r i n g (13 mg). Nhiet dp ndng chay 283-286''C. Phan tich pho ^H-NMR va " C - NlitlR cho thay day la mpt hgp chdt glycosld ma phan aglyeon la [3-sitosterol (so sanh v d i chat 2) va p h i n d u d n g la D-glucose v d i tin hieu cua proton p-anomer d 6H 4,43 (1H, d, J=8,0 Hz) va 6c d 102,24 ppm; cac proton khac cua D- glucoza ndm trong khoang 5H 2,04-2,40 ppm.
Pho cua chat (3) eung t u a n g dong v d i pho c h u i n ciJa p-sitosterol 3-0-P-D-glucopyranosid (daucosterol). Cau true nay eung d u g c khang djnh khi so sanh v d i daucosterol c h u i n trong cung mgt dieu kien s i c ki Idp mdng.
3,4-dihydro-5-hydroxy-7-metoxy-4- (4'-meto xyphenyl)coumarin (4)
Chat (4) la tinh the mau trdng (12 mg), ket tinh trong hon h g p n-hexan/aeelon, Pho *H- WM/? (500 MHz, aceton-de) cho tin hieu cua 4 proton dang AA'XX'doi xdng cua vdng tham hai lan the-para, d 5H 6,8 (2H. dd, J=9.0 va 2.5 Hz, H-3'& H-5') va 7,07 (2H, dd, J=8,5 va 2,5 Hz, H- 2' & H-6'); 2 proton tham khac d vj t r i the meta- vdl nhau d 5H 6.32 (1H, d, J=2,5 Hz, H-6) va 6,24 ( I H , d, J= 2,5 Hz, H-8). Ngoai ra, cdn cd cac tin hieu eua mgt nhdm hydroxyl d AH = 8,9 ppm (Cs) va 2 nhdm metoxy d 6H 3,71 va 3,73 ppm (C4' & C7), cua nhdm metylen d 2,93 (1H, dd, J=16 & 2,.0 Hz. H-3a ) va 3,10 ppm ( I H , dd, J=16 &7,0 Hz, H-3b) va cua metin gan v d i vdng tham CT 4,57 ppm (1H, t, J=7,0 Hz, H-4). Pho
"C-W/WR (125 MHz, aceton-de) cho tin hieu ciia 17C, dc 34,49 (C-4). 37,99 (C-3). 94,42 (C-8), 98,62 (C-6), 106,35 (C-4a), 114,78 (C-3',5'), 128,68 (C-2".6'), 135.12 (C-l'), 154,57 (C-8a), 156,16 (C-5), 159,54 (C-4'), 161.33 (C-7), 167,96 (C-2).
D y a vao s y phan tich pho neu tren. chat (4) phii h g p v d i eau tao 3.4-dihydro-5-hydroxy-7- meloxy-4-(4'-metoxyphenyl)coumarin.
3,4-dihydro-5-hydraxy-7-methoxy4-(4'- methoxyphenyl)coumarin
C h i l (4) lan dau tidn d u g c t i m t h i y trong chi Polygonum.
Quercetin (5) thu d u g c d dang tinh the mSg vang (30 mg), nhi$l dp ndng chay 314-316°C, Ph6 ^H-NMR (500 MHz, aceton-de) cho cdc tin hieu cua 2 proton vdng tham d 5H 6,26 ( I H , d, J=2,0 Hz, H-6) va 5H 6,52 (1H, d, J=2.0 Hz, H-8) d vj tri meta- v d i nhau thudc vdng A. Cdc proton d vdng B, tin hieu d 5H 7,82 (1H, d, J=2,0 Hz, H- 2 ' ) v a d 5 H 7 , 6 9 ( 1 H , d d . J=2,0, 8,5 Hz, H-6') la cua 2 proton d vj tri meta- v d i nhau; proton H- 5' nam canh H-6' nen tin hieu ciia nd xuat hi^n CT 5H 6,99 ( I H , d, J=8,5 Hz, 'H-5'). Pho "C-NMR cho tin hieu cCia 15 carbon. Pho ^H-NMR vS
^^C-NMR ciia chat (5) t u a n g dong v d i eac ph6 t u a n g dng eua quercetin. Dieu nay d u g c khang dinh them khi so sanh chat (5) ciing vdi quercetin chuan trong ciing mgt dieu kign s i c ki Idp mdng.
OH
OH O
Ket luan
B i n g each ket hgp cac p h u a n g ph§p sac ki.
I d phan tren mgt dat cua cay rau m^ ngg {Polygonum perfoliatum L., hg Rau rSm PolygonaceaeJ cac h g p chat stearic acid (1), p- sitoslerol (2), daucosterol (3), 3,4-dihydro-5- hydroxy-7-methoxy-4-(4'-methoxyphenyl) coumarin (4) v a quercetin (5) da d u g c phan lgp. C i u trtc cua Chung da d u g c k h i n g dinh thdng qua nghien c u u ph6 NMR cung n h u so Scinh d6i chung v d i cac c h i t chuan t u a n g u n g . Hgp ch&
TAP CHI DlTQC HQC - 09/2013 (SO 449 NAIM 53)
• Nghien C I J U - Ky thuat
3.4-dihydro-5-hyd roxy-7-methoxy-4-(4'-methoxy phenyl) coumarin (4) lan d i u lien d u g c phan lap t d chi Polygonym.
Tai lieu tham khao
1. D 6 T i t Lai, (2004), NhQvg ciy thuoc va vj thuic Vi$t Nam, Nha xuit b^n Y hoc, tr 111-112.2. Zhang R. L, Sun X. C, Li W. X., Wu L. J., Huang J.. Sun B. H., 2008, "Isolation and idebtification of chemical constituents of Polygonum perfoliatum", Shenyang Pharm. Univ., 105-107.
3. Xingzhong Sun, and Wbert Sneden.
Department of Chemistry, Virginia Commonwealth University, iRichmond, Virginia, USA, 1999,
"Neoflavonoids from Polygonum perfoliatum", Planta Medica. January 12.
4. WANG Qiong, CHEN Li, TIAN Ying, Ll Bin, SUN Qi-Hong, DONG Jun-Xing 2009-03, "Chemical constituents of Polygonum perfoliatum L", Bulletin of the Academy of Military Medical Sciences I.
5. WANG Ding-yong, LU Jiang-Hong, 2004-02;
"Chemical constituents in roots of Polygonum perfoliatum L.", Subtropical Plant Science.
Thanh phan hoa hoc cua cay thanh ngam [Picria fel-terrae Lour.)
Phuffng Thi|n Thuorig^ TrSn Trong Hai^*, Nguyin Tien Dat^, L§ Viet Dung', Nguyen Minh Khdi' ' Vien Dicac lieu
^ Scr Y te tinh San La
^ Vien Hoa Sinh hien, Viin Hdn ldm Khoa hoc vd Cong nghe Viet Nam E-mail: [email protected]
Summary
Our phytochemical studies on the herb Picria fel-terrae (Lour.) collected in Vietnam led to the isolation of six compounds (1-6). Their identification revealed five saponins, including Picfeltarraenin VI, Picfeltarraenin IX, PIcfeltarrenin IA, Picfeltarrenin IB, Picfeltarraenin XI, and a flavonoid apigenin, respectively. This is the first report on the occurence of apigenin in fhe plant Picria fel-terrae Lour..
Keywords: Picria fel-terrae, curcubitanoid saponin, apigenin.
Oat van de
Cdy thanh ngam cd ten khoa hpc la Picria fel-terrae Lour., thugc hp Hoa mom sdi (Scrophulariaceae), la cdy thuoc cd d mien bae Vl$t Nam, mien nam Trung Quoc va mdt so n u d c Odng Nam A '^' ^'. Theo y hpc c6 truyen thanh ngdm ed vj dang, tinh mat, ed tac dung thanh nhigt giai dpc, tieu thung chi thong (lieu sung gi§m dau), khai vi lieu thyc va d u g c diing d ^ chda cSc chdng viem hpng, viem phoi, ho, d^i r i l , ddi ra mau, dau da ddy'^'. Ngudi dan nhi^u n u d c kh^c eung diing thanh ngam d ^ chua b^nh theo y hgc dan gian'^'.
Cic nghidn cuu khoa hgc t r u d c cho biet thanh ngSm cd chda cdc thSnh p h i n chinh gom c6 flavonoid glucosid'^', phenylpropanoid'''^ va
saponin'^'^l Tuy nhien, nghien cuu v ^ tac dung d u g c ly eua thanh ngam cdn it. dac biet la tae dung smh hpc cua cac c h i t nay '^'^"l Nham gdp phan ehung mmh mot each khoa hpc tac dung cua eay thanh ngam. trudc hel chiing toi lien hanh nghien cuu phan ldp va xac dinh cdng thdc mdt so thanh phan hoa hgc l u eay thanh ngam thu hai tai tinh Cao B i n g .
D o i t u ' d n g v a phu'O'ng p h a p n g h i e n CLPU
D o i tira'ng nghien cdu
Toan cay thanh ngam d u g c thu hai tai tinh Cao B i n g vao thang 10/2010. D u g c lieu da TS.
Nguyen Qu6c Huy (BO mdn T h y c vat, Trudng Dgi hpc D u g c Ha Npi) xac djnh ten khoa hgc la
