B GIO DC V O TO

B Y T

TRNG I HC Y DC CN TH

L THANH HUY

NGHIN CU XY DNG CNG THC

VIN NANG CHA CAO DIP C
Houttuynia cordata Thunb.

LUN VN TT NGHIP DC S I HC

NGI HNG DN KHOA HC

Ths. NGUYN TH LINH TUYN

Cn Th - 2011

LI CM N
Em xin gi lng bit n su sc nht n c ThS. Nguyn Th Linh Tuyn
trc tip hng dn, ht lng truyn t kin thc v gip em trong sut qu trnh
thc hin lun vn.
Em xin chn thnh cm n thy DS. Lm Thanh Hng v thy ThS. Chu
Minh Vnh Th tn tnh hng dn v to iu kin cho em thc hin tt lun vn.
Em xin chn thnh cm n Qu Thy C v Anh Ch ky thut vin b mn Cng
Nghip Dc, b mn Dc Liu, b mn Ha Phn Tch - Kim Nghim - c Cht,
b mn Bo Ch h tr dng c, ha cht, trang thit b, gip em ht lng trong
qu trnh thc hin lun vn.
Em xin chn thnh cm n Hi ng chm kha lun tt nghip Dc s i hc
dnh thi gian xem xt, gp v sa cha lun vn c hon thin hn.
Em xin chn thnh cm n Qu Thy C trng i Hc Y Dc Cn Th
du dt, trang b kin thc cho em trong sut nm nm hc va qua.
Xin cm n cc bn oan Trang, Hoa H, Dim Hunh, Mnh Hng, Trng
Tnh, Anh Tun,Thanh Sang, cc bn lp Dc kha 32, cc bn cng nh tr
lun ng vin, chia s, gip trong thi gian qua.
Con xin cm n gia nh - ch da vng chc nht gip con vt qua mi kh
khn th thch trong cng vic v trong cuc sng.
Cn Th, thng 06 nm 2011.

L Thanh Huy

LI CAM OAN
Ti tn L Thanh Huy xin cam oan y l cng trnh
nghin cu ca ring ti. Cc s liu, kt qu nu trong lun
vn l trung thc v cha tng c ai cng b trong bt k
cng trnh no khc.

Cn Th, thng 06 nm 2011.

L Thanh Huy

MC LC
Mc lc.......................................................................................................................i
Danh mc cc k hiu va ch vit tt........................................................................v
Danh mc cc bng..................................................................................................vi
Danh mc cc hnh anh...........................................................................................vii
Danh mc cc s ...............................................................................................viii
T VN ...........................................................................................................1
Chng 1 TNG QUAN........................................................................................3
1.1. TNG QUAN V CY DIP C.................................................................3
1.1.1. c im thc vt....................................................................................3
1.1.2. Phn b sinh thi.....................................................................................3
1.1.3. Thnh phn ha hc.................................................................................4
1.1.3.1. Flavonoid........................................................................................4
1.1.3.2. Tinh du..........................................................................................4
1.1.3.3. Alkaloid..........................................................................................5
1.1.3.4. Cc thnh phn khc.......................................................................5
1.1.4. Tc dng dc l.....................................................................................5
1.1.5. Cng dng...............................................................................................6
1.2. CNG NGH BO CH VIN NANG CNG...........................................7
1.2.1. Khi qut v thuc vin nang..................................................................7
1.2.1.1. nh ngha.......................................................................................7
1.2.1.2. u im ca vin nang cng...........................................................7
1.2.2. Thnh phn vin nang..............................................................................7
1.2.2.1. V nang...........................................................................................7
1.2.2.2. Thnh phn dc cht trong vin nang...........................................8
1.2.3. Quy trnh bao ch thuc vin nang cng t dc liu.............................9
1.3. S LC V PHNG PHP SC K LNG HIU NNG CAO......10
1.3.1. Cc thng s c trng ca qu trnh sc ky..........................................11
1.3.1.1. H s dung lng..........................................................................11

ii

1.3.1.2. H s chn lc...............................................................................11

1.3.1.3. H s bt i xng........................................................................11
1.3.1.4. phn gii..................................................................................11
1.3.1.5. S a l thuyt v hiu lc ct.....................................................12
1.3.2. Mt s ng dng ca phng php HPLC............................................12
1.3.2.1. Phn tch nh tnh........................................................................12
1.3.2.2. Phn tch nh lng.....................................................................12
1.4. GII THIU MT S CH PHM CHA CAO DIP C......................13
1.4.1. An tr vng.........................................................................................13
1.4.2. Helaf......................................................................................................13
1.4.3. Cenditan...............................................................................................14
Chng 2 I TNG V PHNG PHP NGHIN CU..........................15
2.1. NGUYN LIU, CHT CHUN, HA CHT V TRANG THIT B. .15
2.1.1. Nguyn liu v cht chun.....................................................................15
2.1.2. Ha cht.................................................................................................15
2.1.3. Trang thit b.........................................................................................16
2.2. PHNG PHP NGHIN CU................................................................16
2.2.1. Xy dng v thm nh quy trnh nh lng quercetin bng phng
php HPLC......................................................................................................16
2.2.1.1. Kho st cc iu kin sc k ti u.............................................16
2.2.1.2. Kho st phng php x l mu cao Dip c v vin nang cha
cao Dip ca................................................................................................17
2.1.1.3. Thm nh quy trnh nh lng....................................................20
2.2.2. Xy dng cng thc vin nang cha cao Dip c..................................21
2.2.2.1. Thit k cng thc vin nang cha cao Dip c............................21
2.2.2.2. Phng php kho st c tnh ca khi bt thuc v o r ca
vin nang...................................................................................................23
2.2.2.3. Quy trnh iu ch vin nang cha cao Dip c............................24
2.2.3. Xy dng tiu chun kim nghim cho vin nang cha cao Dip c.....24

iii

2.2.3.1. Hnh thc cm quan......................................................................24

2.2.3.2. ng u khi lng................................................................24
2.2.3.3. tan r........................................................................................24
2.2.3.4. m............................................................................................24
2.2.3.5. nh tnh.......................................................................................24
2.2.3.6. nh lng....................................................................................25
Chng 3 - KT QU NGHIN CU...................................................................27
3.1. XY DNG V THM NH QUY TRNH NH LNG
QUERCETIN BNG PHNG PHP HPLC..................................................27
3.1.1. Kho st cc iu kin sc k ti u......................................................27
3.1.2. Kho st quy trnh x l mu cao Dip c v vin nang cha cao
Dip c............................................................................................................28
3.1.3. Thm nh quy trnh nh lng quercetin............................................30
3.1.3.1. Xc nh tnh tng thch h thng...............................................30
3.1.3.2. Thm nh phng php phn tch................................................31
3.2. THIT K CNG THC VIN NANG CHA CAO DIP C...............35
3.2.1. La chn hm lng cao.......................................................................35
3.2.2. La chn hm lng t dc ht...........................................................35
3.2.3. La chn cng thc ti u cho vin nang cha cao Dip c..................36
3.2.3.1. m............................................................................................37
3.2.3.2. Gc chy.......................................................................................37
3.2.3.3. T trng biu kin.........................................................................37
3.2.3.4. Phn b c ht...............................................................................38
3.2.3.5. tan r........................................................................................39
3.3. KIM NGHIM VIN NANG CHA CAO DIP C..............................40
3.3.1. Hnh thc cm quan...............................................................................40
3.3.2. ng u khi lng.............................................................................40
3.3.3. tan r................................................................................................41
3.3.4. m....................................................................................................41

iv

3.3.5. nh tnh................................................................................................41
3.3.5.1. Bng phn ng ha hc.................................................................41
3.3.5.2. Bng SKLM..................................................................................41
3.3.6. nh lng.............................................................................................42
3.4. XY DNG TIU CHUN VIN NANG CHA CAO DIP C...........42
3.4.1. Tiu chun k thut...............................................................................42
3.4.2. Kt qu kim nghim.............................................................................46
Chng 4 - BN LUN.........................................................................................47
4.1. V xy dng v thm inh quy trnh inh lng quercetin trong cao Dip c
v vin nang cha cao Dip c............................................................................47
4.2. V nghin cu xy dng cng thc cho vin nang cha cao Dip c...........47
4.3. V xy dng tiu chun kim nghim cho vin nang cha cao Dip c.......48
Chg 5 - KT LUN V KIN NGH................................................................49
5.1. Kt lun........................................................................................................49
5.2. Kin nghi......................................................................................................49
TI LIU THAM KHO.......................................................................................51

DANH MC CC K HIU V CH VIT TT

AOAC

Hip hi cc nh ho hc phn tch quc t

(Association of Official Analytical Chemists)

CHCl3

Chloroform

CH3CN

Acetonitril

DVN

Dc in Vit Nam

dtv

T trng trc va p

dsv

T trng sau va p

HPLC

Sc k lng hiu nng cao

(High Performance Liquid Chromatography)

MeOH

Methanol

RSD

lch chun tng i (Relative Standard Deviation)

SD

lch chun (Standard Deviation)

SKLM

Sc k lp mng

STT

S th t

TB

Trung bnh

tR

Thi gian lu

TT

Thuc th

UV

T ngoi (Ultra Violet)

Vtv

Th tch trc va p

Vsv

Th tch sau va p

DANH MC CC BNG
Trang
Bng 2.1. Cc ha cht, t dc s dng trong nghin cu.....................................15
Bng 2.2. Cc thit b dung trong nghin cu.........................................................16
Bng 2.3. Phng php pha cc dung dch c nng thp hn............................17

vi

Bng 2.4. Cc cng thc thit k ti u cng thc vin nang.................................22

Bng 2.5. nh tnh vin nang cha cao Dip c bng phn ng ha hc...............25
Bng 3.1. Kt qu kho st tnh tng thch h thng trn 6 ln tim mu chun...30
Bng 3.2. Kt qu kho st tnh tng thch h thng trn 6 ln tim mu th.......30
Bng 3.3. Kt qu kho st tnh tuyn tnh phng php nh lng bng HPLC. .33
Bng 3.4. Kt qu kho st chnh xc phng php nh lng bng HPLC.....34
Bng 3.5. Kt qu kho st ng ca phng php nh lng bng HPLC......34
Bng 3.6. Cc cng thc thit k xc nh hm lng cao thch hp.................35
Bng 3.7. m cc cng thc C1, C2, C3, C4.....................................................35
Bng 3.8. Cc cng thc thit k xc nh hm lng t dc ht.....................35
Bng 3.9. m cc cng thc H1, H2, H3, H4....................................................36
Bng 3.10. Cc cng thc thit k ti u ha cng thc vin nang.........................36
Bng 3.11. m cc cng thc kho st (T1 T10)............................................37
Bng 3.12. Gc chy cc cng thc kho st (T1 T10)........................................37
Bng 3.13. T trng biu kin ca cc cng thc kho st (T1 T10)...................37
Bng 3.14. Phn b c ht ca cc cng thc kho st (T1 T10).........................38
Bng 3.15. r cc cng thc kho st (T1 T10)..............................................39
Bng 3.16. Kt qu kim nghim ng u v khi lng.................................40
Bang 3.17. Kt qua o ra....................................................................................41
Bang 3.18. Kt qua o m..................................................................................41
Bng 3.19. Kt qu nh tnh vin nang cha cao Dip c......................................41
Bng 3.20. Kt qu kim nghim vin nang Dip c..............................................46

DANH MC CC HNH NH
Trang
Hnh 1.1. Cy Dip c...............................................................................................3
Hnh 1.2. Cng thc cu to ca quercitrin v isoquercitrin.....................................4

vii

Hnh 1.3. S cu to chung ca mt h thng HPLC.........................................10

Hnh 1.4. Cach xc nh thi gian cht (t0) v thi gian lu (tR).............................11
Hnh 1.5. Cch xc nh s a l thuyt.................................................................12
Hnh 1.6. Ch phm An Tr Vng.........................................................................13
Hnh 1.7. Ch phm Helaf......................................................................................13
Hnh 1.8. Ch phm Cenditan.................................................................................14
Hnh 2.1. Phng php xc nh gc ngh ca bt, ht thuc.................................24
Hnh 3.1. Sc k 3D mu th bc sng t 250nm 450nm..........................27
Hnh 3.2. Sc k contour mu th......................................................................27
Hnh 3.3. Sc k cc dch thy phn quercetin sau 1h, 2h v 3h........................28
Hnh 3.4. Sc k mu th trc v sau khi loi tp bng SPE t ch.................29
Hnh 3.5. Sc k mu chun...............................................................................31
Hnh 3.6. Sc k mu th...................................................................................31
Hnh 3.7 Sc k mu trng.................................................................................31
Hnh 3.8. Sc k mu dung mi.........................................................................32
Hnh 3.9. Sc k mu dung dich nc acid phosphoric 0,2%............................32
Hnh 3.10. Kt qu chng ph mu chun, mu dung mi, mu t dc v mu
dung dch nc acid phosphoric 0,2%.....................................................................32
Hnh 3.11. Biu tnh tuyn tnh ca phng php nh lng bng HPLC........33
Hnh 3.12. Biu phn b c ht ca cc cng thc t T1 n T5.......................38
Hnh 3.13. Biu phn b c ht ca cc cng thc t T6 n T10.....................39
Hnh 3.14. Kt qu nh tnh bng SKLM vin nang cha cao Dip c..................41

DANH MC CC S
S 1.1. Tm tt quy trnh sn xut vin nang cng..............................................9
S 2.1. Tm tt quy trnh x l mu th.............................................................19

T VN
Quay v vi thin nhin l xu hng ca th ky XXI. Trong lnh vc Dc, cc
thuc c ngun gc t dc liu ngy cng c quan tm nhiu hn. Trc y,
ngi ta thng hiu ch c mt s nc phng ng nh Trung Quc, Vit Nam,
mi c nhiu sn phm t thin nhin. Thc t, theo T Chc Y T Th Gii
(WHO) hin c ti 60% dn s s dng cc sn phm ny t cc nc chu nh
n , Thi Lan n cc nc Chu Phi v M La Tinh. Trong c nhiu nhm,
loi thuc chim t trng cao: chng hn chit xut thc vt cha ph i tin lit
tuyn lnh tnh c ti 100 bit dc, chim ti 25 36% (Php, c) 3,5 10,5%
(Italia, Ty Ban Nha) so vi tng s dc phm cha bnh ny. [32]
Vit Nam l nc nhit i gi ma, ng thc vt phong ph l tin tt
pht trin cc sn phm t dc liu. Ngun dc liu di do l nn tng cho vic
hnh thnh cc c s sn xut thuc t dc liu. Theo thng k ca B Y t, c
nc hin c hn 300 c s sn xut thuc t dc liu trong nc, trong c 10
c s ng dc t chun thc hnh tt sn xut thuc ca WHO. Nhiu cng ty
dc cng c thng hiu ring t th mnh ca cy dc liu. [31]
Trong cac loai dc liu ph bin, Dip c l loi rau n quen thuc hng ngy
c nhn dn ta s dng lm thuc vi nhiu tc dng: tr to bn, tr, mn nht,
l nga,.Tuy nhin c mt s bt tin trong vic s dng dc liu ny nh kh
n v khng th n vi lng ln. Mt khc, vic s dng dc liu nh thc phm
hay cc sn phm khng c chun ha v kim sot cht ch s nh hng ln
n tnh an ton v hiu qu tr liu. V vy, ti nghin cu xy dng cng thc
vin nang cha cao Dip c c thc hin vi nhng mc tiu c th nh sau:
1. Xy dng v thm nh quy trnh nh lng quercetin trong cao Dip c v

vin nang cha cao Dip c.

2. Xy dng cng thc vin nang cha cao Dip c.
3. Xy dng tiu chun kim nghim cho vin nang bo ch c.

ngha ca ti:
1. Gp phn nghin cu v cung cp c s l lun bo ch cc ch phm t
cao dc liu.
2. L c s cho cc nghin cu sau ny trn quy m pilot v cng nghip.

Chng 1 TNG QUAN

1.1. TNG QUAN V CY DIP C:
Tn khc: L gip, rau gip c, tp thi, ng tinh tho, rau vn.
Tn khoa hc: Houttuynia cordata Thunb.
H l gip: Saururaceae.
1.1.1. c im thc vt: [6], [13], [16]

Cy tho, sng lu nm, cao 20 40cm, thn

ngm mc b ngang trong t, mu trng, hi c
lng, bn r cc mu. Thn ng nhn, mu lc
hoc tm . L mc so le, hnh tim, u nhn,
mt trn mu lc sm, mt di mu tm, hi c
lng mc theo gn l ca c hai mt, gn chnh 7;
cun l di, c b, l km c lng mp.
Cm hoa mc ngn thn thnh bng di 2
2,5cm, mang nhiu hoa nh mu vng nht; tng
bao gm 4 l bc mu trng nom nh mt chic

Hnh 1.1. Cy Dip c.

hoa ring l; hoa khng c bao hoa; nh 3. Qu

nang m nh; ht hnh tri xoan nhn. Ton thn v ra c mi tanh nh mi c.
Ma hoa qu: thng 5 7.
1.1.2. Phn b sinh thi: [13], [16]
Dip c phn b ch yu vng nhit i v cn nhit i chu , t Nht
Bn, Trung Quc n Vit Nam, Lo, n v cc nc ng Nam A khc.
Vit Nam, cy mc hoang di cc tnh min ni, trung du v ng bng. Cy cn
c trng nhiu ni lm rau v lm thuc. Dip c thuc cy u m v hi
chu bng, thng mc t m, nhiu mn dc theo cc b khe sui, mng nc
trong thung lng v vng ng bng. Cy sinh trng gn nh quang nm, mnh
nht trong ma xun h, c hoa qu hng nm trn nhng cy khng b ngt ngn
v hi l thng xuyn, c kh nng ti sinh chi mnh t thn r.

1.1.3. Thnh phn ha hc:

1.1.3.1. Flavonoid:
Cc flavonoid ng ch l: afzelin, phloretin, rutin, quercitrin (quercetin 3rhamnosid), isoquercitrin (quercetin 3-glucosid), dioxy flavonon. [6]

quercitrin
isoquercitrin
Hnh 1.2. Cng thc cu to ca quercitrin v isoquercitrin.
Cc flavonoid khc l dng glycosid ca quercetin nh: [12], [18], [19], [27]
- Quercetin-3-O--D-galactosid-7-O-beta-D-glucosid
- Quercetin-3-O--D-galactopyranosid (hyperin)
- Quercetin-3-O--L-rhamnopyranosyl-7-O--D-glucopyranosid
- Quercetin-3-O--L-arabinofuranosid (avicularin)

Trong quercitrin, isoquercitin v phloretin c coi l nhng hp cht c tc

dng chng ung th v tc dng ngn chn gc t do. [18], [19]
1.1.3.2. Tinh du:
Ton thn Dip c cha tinh du, y l thnh phn lm cho dc liu c mi
c bit. Thnh phn ch yu l nhm aldehyde v cc dn cht ceton nh methyln-nonylceton, 1-decanal, 1-dodecanal l nhng cht khng c tc dng khng
khun, cht c tc dng khng khun l 3-oxododecanal, y l thnh phn chnh
ca tinh du nhng khng bn v d b phn hy khi chng ct. Nhm terpen bao
gm cc cht: -pinen, camphen, myrcen, limonen, linalol, bornyl acetat, geraniol
v caryophylen. [6]
Ngoi ra tinh du Dip c cn cha acid hu c: acid caprinic, laurinaldehyde,
benzamid, acid hexadecanoic, acid decanoid, acid palmitic, acid linoleic, acid oleic,
acid stearic,lipid v vitamin K. [6], [30]
1.1.3.3 Alkaloid:

T Dip c phn lp c -sitosterol [14], su alkaloid c hot tnh sinh

hc l aristolactam B(1), piperolactam A(2), aristolactam A(3), norcepharadione
B(4), cepharadione B(5) v splendidine(6) cng c tm thy trong dch chit
methanol phn trn mt t ca cy Dip c. [23]
1.1.3.4.Cc thnh phn khc:
Nc: 91,5%, protid: 2,9%, glucid: 2,7%, lipid: 0,5%, cellulose:1,8%, Calci:
0,3mg, Kali: 0,1mg, tin vitamin A: 1,26mg, vitamin C: 68mg (trong 100g rau Dip
c ti). [13]
Ngoi ra Dip c cn cha St (Fe), Magnesi (Mg), Mangan (Mn), khng tm
thy cc kim loi nng nh Arsen (As), Cadimi (Cd), Chrom (Cr), hoc Ch (Pb)
trong mu phn tch. [20]
1.1.4. Tc dng dc l:
Tc dng khng virus: [21]
Theo Kyoko Hayashi v cng s Dip c c ch trc tip virus herpes chng 1
(HSV-1), virus cm, virus gy suy gim min dch mc phi ngi loi 1 (HIV-1)
m khng biu hin c tnh, nhng khng chng li poliovirus v coxsackie virus.
Mc gim virus lin quan n thi gian x l bng thuc. Ba thnh phn chnh
c tc dng l: xeton methyl n-nonyl, aldehyde lauryl, v aldehyde capryl.
Tc dng chng ung th mu: [17]
Cng trnh nghin cu ca Chang Jung-San v cng s, mc ch nh gi
tc dng chng ung th mu ca H.cordata v Bidens pilosa. Nm dng t bo ung
th mu, theo th t l L1210, U937, K562, Raji v P3HR1 c nui cy vi cht
chit xut ca H. cordata v Bidens pilosa. Kt qu cho thy chit xut t cc dc
liu trn c tc dng ngn chn 5 dng t bo ny.
Tc dng chng vim tuyn v: [29]
Cng trnh nghin cu c thc hin ti Trng i hc Th y, thuc Vin i
hc Nng nghip Zbejiang, Hangzbou, Trung Quc. T Houttuynia cordata, cc
nh nghin cu chit xut c cht Houttuynia, sau cho Houttuynia phn

ng vi bisulphat natri c c cht Houttuynin bisulphat natri. T cht ny, h

ch bin c mt dung dch tim vo v iu tr bnh vim tuyn v b.
Th nghim trn 52 trng hp vim tuyn v b c chia lm 2 nhm:
nhm 1 tim 80mg dung dch H. bisulphat Na v nhm 2 tim 800.000 n v
Penicillin kt hp vi 1g Streptomycin. Kt qu cho thy 88,2% vim tuyn v
iu tr khi vi H.bisulphat Na, so vi 90% c cha lnh vi Penicillin +
Streptomycin. Nh vy, khng c s khc bit ng k trong iu tr vim tuyn v
cp tnh gia 2 nhm trn.
Cc tc dng khc:
Dip c cn c tc dng khng khun [26], khng vim [22], chng oxy ha
[28]. Dch chit nc Dip c dng iu tr cho bnh nhn vim xoang mn tnh v
polyp mi [25].
1.1.5. Cng dng: [6], [13], [16]
Dip c c dng tr to bn, tr, mn nht, l nga, tr con ln si, vim phi
hoc phi c m, au mt hoc au mt do nhim trc khun m xanh, vim
rut, kit l, b tiu tin, kinh nguyt khng u.
Trung Quc, mt hp cht c hot tnh khng khun c phn lp t cy
dip c v bo ch thnh thuc vin v thuc tim tr bnh nhim khun.
Nht Bn, thn r Dip ca c trong thnh phn ca mt s ch phm thuc
dng cha mt s bnh ca ph n.

1.2. CNG NGH BO CH VIN NANG CNG:

1.2.1. Khi qut v thuc vin nang:[2]
1.2.1.1. nh ngha:
Thuc nang l dng thuc ung cha mt hay nhiu hot cht trong v nang
cng hay mm vi nhiu kiu dng v kch thc khc nhau. V nang c lm t
gelatin v c th c thm cc cht ph gia khng gy c hi cho c th ngi.
Thuc nang cng c v nang gm hai phn hnh tr lng khch vo nhau, mi phn
c mt u kn, u kia h. Thuc ng trong nang thng dng rn (bt hay
cm).
1.2.1.2. u im ca vin nang cng:
Vin nang l dng vin d ung, d nut v c mu sc phong ph hn dng
vin nn.
Dc cht ng vo vin nang c th nhiu dng: bt, cm, vi ht, vi nang,
vin nang nh, vin nn hoc phi hp cc dng trn trong cng mt v nang.
S phi hp ny c th gip cch ly cc thnh phn tng k hoc iu ch
vin nang phng thch ko di bng cch phi hp cc vi ht hoc vi nang phng
thch dc cht ti nhiu thi im v v tr khc nhau trong ng tiu ha d dng.

So vi vin nn, vin nang l dng thuc tng i d nghin cu xy dng

cng thc. D trin khai sn xut cc quy m khc nhau, c th s dng cc my
ng nang th cng trong quy m nh hoc cc my ng nang bn t ng v t
ng trong quy m sn xut ln.
Vin nang l dng vin t gp phi cc vn v sinh kh dng do khi thuc
trong nang khng b nn cht nn vin d r hn vin nn.
1.2.2. Thnh phn vin nang:[2]
1.2.2.1. V nang:
V nang c ch to t nguyn liu chnh l gelatin, cc cht mu, cht to
c nh titan dioxyd v cc cht ph gia khc.
V nang cng c th ch to t dn cht cellulose, loi v nang ny t c s
dng v tan km v gi thnh cao.

1.2.2.2. Thnh phn dc cht trong vin nang:

Xy dng cng thc cho vin nang
Khi thuc (ht, bt) ng vo nang phi c 2 tnh cht c bn l trn
chy, tnh chu nn. Cc thuc tnh ny c th thay i nht nh ty thuc vo thit
b ng thuc vo nang.
Cn lu l cc dc cht c tnh ht m cao c kh nng lm mm v nang,
cc dc cht c tnh kim cao hoc acid cao cng c th lm hng v nang.
tng lu tnh v kh nng chu nn ca khi thuc, c th p dng phng
php xt ht kh hoc xt ht t.
Kch thc ca ht nn ph hp c th m bo ht chy u vo nang ng
thi hn ch c sai s khi lng thuc trong nang.
Cc t dc thng thng dng iu ch khi bt gm:
T dc n: Cc loi t dc n dng trong vin nn nh tinh bt, lactose,
dicalci phosphat u c th c dng trong vin nang. Cc loi tinh bt dp thng
nh tinh bt tin gelatin ha, tinh bt phun sy c th c dng gia tng lu
tnh v tnh chu nn ca khi ht.
T dc trn: t dc trn gip cho ht chy u. S trn chy ca khi ht
hoc bt cn thit cho tt c cc my ng nang khc nhau.. trn chy c bit
cn thit trong trng hp ng thuc theo nguyn tc a phn liu. Cc t dc
trn thng dng l Mg stearate, talc, tinh bt bp,
T dc chng dnh: cc t dc chng dnh va c tc dng lm tng lu tnh
ca khi bt (hoc ht) va trnh s kt dnh ca bt thuc ln cc b mt kim loi.
T dc r: t dc r c th khng cn thit trong trng hp ng bt khng
nn vo nang. Trong trng hp c xt ht hoc trong trng hp c nn p (my
c a phn liu hoc vt phn liu) th nn c t dc r gip thuc phng thch
nhanh. Nn s dng cc t dc siu r c th chn c kiu nang nh.

1.2.3. Quy trnh bao ch thuc vin nang cng t dc liu: [1],[2]
iu ch khi thuc
V nang
ng nang

Lau nang nh bng

ng gi

Thnh phm

S 1.1. Tm tt quy trnh sn xut vin nang cng.

iu ch khi thuc: thng thng iu ch khi thuc bng phng php xt
ht kh hoc xt ht t. i vi dc liu thng tin hanh nh sau:
+

Chit kit hoat cht t dc liu bng phng phap va dung mi thich hp.

X ly dich chit thu c cao c hoc cao kh.

Do bt thuc lam t dc liu hut m rt manh nn phai khao sat va phi

hp cao dc liu vi ta dc hut m, ta dc n theo ty l phu hp nhm gi n
inh va bao quan khi bt thuc.

Ep cm, phi hp ta dc trn chay thich hp.

Sa hat va kim tra cht lng hat.

ng nang: Trong trng hp iu ch mt lng nh vin nang dng cho
mt s bnh nhn hoc th nghim lm sng cc thuc mi, khi thuc c th c
ng vo nang bng tay (khng dng thit b). Trong sn xut c th s dng cc
my ng nang th cng, bn t ng v my ng nang t ng ty theo quy m
sn xut khc nhau.

10

Lau nang nh bng: sau khi ng thuc vo nang, cc vin nang cn c

loi bi v nh bng trc khi ng gi. Ty theo iu kin trang thit b c sn,
c th p dng cc phng php: loi bi bng phng php th cng, loi bi v
nh bng trong ni bao, loi bi bng h thng lau v nh bng vin.
1.3. S LC V PHNG PHP SC K LNG HIU NNG CAO: [9]

Sc k lng hiu nng cao i khi cn c gi l sc k lng p sut cao

(High-Pressure) l k thut phn tch da trn c s ca s phn tch cc cht trn
mt pha tnh cha trong ct, nh dng di chuyn ca pha ng lng di p sut
cao. Sc k lng da trn c ch hp ph, phn b, trao i ion hay loi c l ty
thuc vo loi pha tnh s dng.
Ngy nay, HPLC v ang c s dng trong nhiu lnh vc phn tch ha
hc ni chung cng nh trong kim tra cht lng thuc v phn tch sinh dc hc
ni ring. Trong phn tch thuc bng phng php sc k, phn ln cc dc in
u s dng sc k phn b.

Hnh 1.3. S cu to chung ca mt h thng HPLC.

1.3.1. Cc thng s c trng ca qu trnh sc k:

11

1.3.1.1. H s dung lng k (Capacity factor):

k'=

t 'R t R t0
=
t0
t0

Trong :
k': h s dung lng.
tR: thi gian lu.
tR: thi gian lu hiu chnh.
t0: thi gian cht.

Hnh 1.4. Xc nh thi gian cht (t0)

Cn chn ct, pha ng,sao cho k

v thi gian lu (tR).

nm trong khong ti u: 1 k 8.
1.3.1.2. H s chn lc :

k ' B t RB
=
k ' A t RA

Quy c y B l cht b lu gi mnh hn A nn > 1.

tch ring 2 cht thng chn 1,05 2,0.
1.3.1.3. H s bt i xng:

As =

W1/ 20
2f

Trong :
W1/20: rng nh 1/20 chiu cao.
f: khong cch t ng cao ca nh n chn trc ca nh 1/20 chiu cao.

1.3.1.4. phn gii Rs:

Rs =
Trong :

2(t RB t RA )
t t
= 1,18 RB RA
WB + WA
W1/ 2 B + W1/ 2 A
TRB, tRA: thi gian lu ca 2 pic lin k nhau (B v A).
WB, WA: rng pic o y cc pic.
W1/2B, W1/2A: rng pic o na chiu cao pic.

1.3.1.5. S a l thuyt v hiu lc ct:

12

Hiu lc ct c o bng thng s: s a l thuyt N ca ct.

t R2
t R2
N = 16 2 = 5,54 2
W
W1 / 2
Trong :
W: chiu rng o y pic.
W1/2: chiu rng pic o na chiu
cao pic.

Hnh 1.5. Cch xc nh s a l thuyt.

1.3.2. Mt s ng dng ca phng php HPLC:
1.3.2.1. Phn tch nh tnh:
nh tnh cc hp cht l mt phn ct li ca bt c mt phng phap HPLC
no. Phn tch nh tnh bng HPLC ch yu thc hin bng 3 cch sau y:

Da vo cc d liu v s lu.

Phn tch nh tnh cc cht thu c t HPLC quy m phn tch.

Phn tch ph nghim online cc nh HPLC.

1.3.2.2. Phn tch nh lng:

Nguyn tc: vic nh lng cc cht bng HPLC c th da vo s so snh
chiu cao ca nh hay so snh din tch nh ca cht cn xc nh vi mt hay
nhiu mu chun bit trc nng .
Cc phng php nh lng:

Phng php quy v 100% din tch.

Phng php dng chun ngoi.

Phng php dng chun ni.

Phng php thm cht chun.

1.4. GII THIU MT S CH PHM CHA CAO DIP C:

13

1.4.1. An tr vng: Cng ty C phn sn xut v Thng mi Hng Bng.

CNG THC:
Hm lng trong mt vin:
Cao Dip c
Cao ng quy
Magie carbonat
Rutin
Curcumin

450 mg
150 mg
108 mg
25 mg
10 mg

Hnh 1.6. Ch phm An Tr Vng.

CNG DNG:
H tr iu tr v gip phng nga bnh tr, ci thin cc triu chng ca bnh
tr (chy mu, sa bi tr, au rt, nga...) v cc bin chng xut huyt ca bnh tr
(sa trc trng, vim nt hu mn...).
H tr iu tr v phng nga to bn.
Gip bo v v tng sc bn ca tnh mch, tng cng sc khe tnh mch v
ng tiu ha.
1.4.2. Helaf: Cng ty c phn dc Hu Giang.
CNG THC:
Hm lng trong mt vin:
Cao kh Dip c
210 mg
Cao kh Rau m
45 mg
Kollidon CL - M, sp ong trng,
du nnh, v nang gelatin

CNG DNG:

Hnh 1.7. Ch phm Helaf.

Helaf c tc dng h tr iu tr tr, to bn v kit l.

Helaf gip gii nhit, thng tiu, mt gan, gii c, khng vim, gip vt
thng chng lnh v mau ln da non, tng cng h min dch ca c th.
1.4.3. Cenditan: Cng ty c phm dc phm 3/2.

14

CNG THC:
Hm lng trong mt vin:
Cao dip c
Bt rau m

75mg
300mg

CNG DNG:
Tr to bn, tr.
Gii nhit, thng tiu, mt gan, gii c.
Hnh 1.8. Ch phm Cenditan.

Chng 2 I TNG V
PHNG PHP NGHIN CU
2.1. NGUYN LIU, CHT CHUN, HA CHT V TRANG THIT B:

15

2.1.1. Nguyn liu v cht chun:

Cao Dip c c cung cp bi cng ty TNHH ADC ti Mn, KV Thi Hng,
phng Long Hng, qun Mn, Thnh ph Cn Th. S l: SE-HCO1-0210.

Quercetin chun do B Y T Vin kim nghim TP.HCM cung cp, s l

104050609, ham lng 91,34% C15H10O7, nc 8,51%.
2.1.2. Ha cht:
Bng 2.1. Cc ha cht, t dc s dng trong nghin cu.
STT
1
2
3
4
5
6
7
8
9
10
11
12
13
12

Tn nguyn liu, ha cht

Acetonitril
Acid chlohyric
Acid formic
Acid phosphoric
Chloroform
Ethanol 96%
Ethyl acetat
Magnesi
Magnesi carbonate
Methanol
Natri hydroxyd
Natri sulfat
St (III) clorid
Talc

Ngun gc
Merck
Trung Quc
Trung Quc
Merck
Trung Quc
Vit Nam
Trung Quc
Trung Quc
Vit Nam
Merck
Trung Quc
Trung Quc
Trung Quc
M

Tiu chun
Phn tch HPLC
Tinh khit PA
Tinh khit PA
Phn tch HPLC
Tinh khit PA
TCCS
Tinh khit PA
Tinh khit PA
TCCS
Phn tch HPLC
Tinh khit PA
Tinh khit PA
Tinh khit PA
USP 30

2.1.3. Trang thit b:

Bng 2.2. Cc thit b dng trong nghin cu.
STT
1
2
3
4
5
6
7
8

Tn thit b
Bp in
B siu m
Cn phn tch
My o r
My st ht v sa ht
My trn
T sy
My soi UV 2 bc sng

S hiu
JK 071
Elma
Ep 214C-Ohaus
Pharma test
FGS
Misuko JBJ103
Memmerk
ENF-2400C/EF

Ngun gc
Nht
c
M
c
c
Nht
c
M

16

9
10

My HPLC
Cn sy m hng ngoi

Hitachi L200
MX-50

Nht
M

2.2. PHNG PHP NGHIN CU:

2.2.1. Xy dng v thm nh quy trnh nh lng quercetin bng phng
php HPLC:
2.2.1.1. Kho st cc iu kin sc k ti u:
Tin hnh chy sc k trn quercetin chun tm cc iu kin sc k ti u.
Chy sc k vi mu th iu kin sc k thm d, tin hnh thay i cc
iu kin sc k nh chnh (tng ng vi quercetin) t cc thng s sc k.
Chun b mu chun: cn chnh xc 25mg chun i chiu quercetin cho vo
bnh nh mc 50ml, thm 30ml methanol, siu m trong 10 pht v b sung
methanol n vch, lc u c dung dch m c nng 500g/ml.
T dung dch m, tin hnh pha cc dung dch c nng thp hn nh sau: ly
chnh xc mt lng th tch dung dch m cho vo bnh nh mc, thm methanol
va thu c dung dch c nng theo yu cu. Cc s liu pha mu c trnh
by trong bng 2.3.
Bng 2.3. Phng php pha cc dung dch c nng thp hn.
Dung dch m

STT
1
2
3
4
5
6

Bnh nh mc

(ml)
(ml)
1
10
2
10
3
10
4
10
5
10
6
10
Chun b mu th: tin hnh theo mc 2.2.1.2.

Nng dung dch thu c

(g/ml)
50
100
150
200
250
300

2.2.1.2. Kho st phng php x l mu cao Dip c v vin nang cha cao
Dip c nh lng quercetin:
Tin hnh chit v theo di:
-

Qu trnh chit quercetin t dch thy phn cao Dip c.

17
-

Thi gian phn ng thy phn xy ra hon ton: dch thy phn sau
mi 1 gi c chit kit Quercetin bng CHCl3, theo di cc dch thy
phn sau 1 gi, 2 gi, 3 gi,...

Qu trnh loi tp.

Theo di bng thuc th FeCl3 trong cn v sc k lp mng vi h dung mi

Chloroform Ethyl acetat Acid formic (5:4:1). Pht hin bng n UV bc
sng 254 nm, nhng thuc th FeCl3 trong cn.
Quy trnh chun b mu th cao Dip c:
Cn chnh xc khong 1 g cao Dip c, ha tan vi 10 ml ethanol 25%, lc,
chuyn vo bnh lng gn, thm vo 20 ml chloroform, lc u 5 pht, rt b dch
chloroform. Dch nc cn li ha tan vi 10 ml dung dich HCl 10%/cn 25% un
hi lu. Dch thy phn thu c lc vi vi 20 ml chloroform trong 5 pht, chuyn
vo bnh lng gn yn 60 pht, rt ly dch chloroform. Lp li 3 ln. Gp dch
chloroform lc vi vi 20 ml nc. Rt dch chloroform lc qua Na 2SO4 sau em
c n dch CHCl3 m c (A1).
Dch A1 cho qua ct SPE t ch loi tp. Cho 20 ml chloroform qua ct SPE
loi tp phn cc, tip theo cho 40ml hn hp CHCl3: EtOAc 6:4 ly phn
on cha quercetin. C phn on cha quercetin n cn (A2).
Ha tan cn A2 trong methanol ri cho vo bnh nh mc 10ml, b sung
th tch, lc s b. Sau lc qua mng lc 0,45m, b vi ml dung dch u trc
khi bm vo h thng.
Quy trnh chun b mu th vin nang cha Cao dip c:
Cn 20 nang, tnh khi lng trung bnh ca bt thuc trong nang, nghin thnh
bt mn. Cn lng bt thuc tng ng vi 1g cao Dip c, ha tan trong 10 ml
cn 25%, lc thu ly dch lc, ra cn vi khong 2 ml cn 25%, thm vo 20 ml
chloroform, lc u 5 pht, chuyn vo bnh lng gn, rt b dch chloroform. Dch
nc cn li ha tan vi 10 ml dung dich HCl 10%/cn 25% un hi lu. Dch thy
phn thu c lc vi 20 ml chloroform trong 5 pht, chuyn vo bnh lng gn
yn 60 pht, rt ly dch chloroform. Lp li 3 ln. Gp dch chloroform lc vi 20

18

ml nc. Rt dch chloroform lc qua Na2SO4 sau em c n dch CHCl3 m

c (A1).
Cc bc cn li x l nh i vi mu th cao Dip c.
Cao Dip c
(Vin Dip c)
Ha tan vo cn 25%, lc
Lc vi CHCl3, gn b lp CHCl3
Dch cn 25%
HCl 10% / cn 25%
Thy phn 3h
Dch thy phn
Lc vi CHCl3 3 ln

Dch CHCl3
Lc vi H2O, gn b lp nc. Lc qua Na2SO4
C n dch CHCl3 m c
Dch CHCl3 m c
Ct SPE t ch

Dch SPE cha

Quercetin
C n cn

Cn

S 2.1. Tm tt quy trnh x l mu th.

19

2.1.1.3. Thm nh quy trnh nh lng:

Tnh tng thch h thng:
Tnh tng thch h thng ca quy trnh phn tch c xc nh trn gi tr v
lp li ca cc thng s k thut ca h thng khi tin hnh thc hin quy trnh
trn mu chun v mu th vi 6 ln tim mu lin tip.
- Tin hnh:

Chun b mu th v mu chun.
Chy sc k 6 ln vi mu chun v mu th.

Kho st cc thng s: thi gian lu, din tch nh.

X l thng k cc gi tr thu c.
- Yu cu:
RSD 2%.
Thm nh quy trnh:
Tnh c hiu
- Tin hnh:

Chun b mu th v mu trng (l mu ch gm t dc),

mu dung mi pha mu, mu dung dch nc acid phosphoric

0,2%.
- Yu cu:

Tin hnh chy sc k.

Mu trng, mu dung mi, mu dung dch nc acid
phosphoric 0,2% khng c cc pic trng vi nh chnh
(tng ng vi quercetin).

Tnh tuyn tnh

- Tin hnh:

Chy sc k v xc nh hm lng 6 dung dch chun vi

nng khc nhau.

Xy dng ng chun v xc nh R2.
Thm nh li phng trnh ng chun bng phng php

- Yu cu:

thng k.
R2 0,99.

20

chnh xc
- Tin hnh:

X l 6 mu th nng bng nhau.

Xc nh hm lng quercetin trong mi mu th.

- Yu cu:

Tnh hm lng trung bnh v RSD%.

RSD nm trong gii hn cho php ca AOAC.

ng
- Tin hnh:

Thm lng cht chun quercetin tng ng vi khong

80%, 100%, 120% hm lng quercetin c trong mu th.

nh lng tm li hm lng hm lng quercetin thm

- Yu cu:

vo xc nh t l phc hi.
T l phc hi trong gii hn cho php ca AOAC.

2.2.2. Xy dng cng thc vin nang cha cao Dip c:

Qu trnh nghin cu nh sau: thit k cng thc to ht kho st c tnh
ca ht ng nang th r la chn cng thc ch phm.
Cng thc c thit k da theo phng php c in ca Gauss Geidell. [8]
2.2.2.1. Thit k cng thc vin nang cha cao Dip c:
La chn hm lng cao thch hp:
Cao dc liu c kh nng ht m rt ln, hm lng cao cng nhiu th kh
nng ht m cng mnh. V vy, chn hm lng cao thch hp sao cho gim bt
lng t dc s dng m vn m bo m thch hp cho vic ng nang l iu
cn thit. thc hin iu ny chng ti tin hnh nh sau:
- Thit k cc cng thc vi hm lng cao khc nhau, gi nguyn t l t dc

ht ti a c th s dng trong cng thc.

- Tin hnh to ht v kho st m lc cn bng tm hm lng cao thch
hp.
- Hm lng cao thch hp l hm lng cao cao nht v c m bt thuc lc

cn bng 5%.

21

La chn hm lng t dc dnh:

Cao dc liu c dnh kh ln, v vy c th lm m bng nc v tin hnh
st ht, tuy nhin t l bt mn thng cao. Do vy chng ti chn PVP K30 lm t
dc dnh vi hm lng l 2%.
La chn hm lng t dc ht:
Xy dng cng thc vi hm lng t dc ht ln lt l 0%, 5%, 10%, 15%,
xc nh m lc cn bng ca khi bt thuc.
Chn hm lng t dc ht sao cho m lc cn bng < 5%.
La chn t cn li v ti u cng thc vin nang:
Sau khi chn hm lng cao v cc loi t dc, chng ti tin hnh thit k 10
cng thc vi 2 loi t dc n A v B c t l: 3:1, 2:1, 1:1, 1:2,1:3; t l t dc
ht gi tr ti a v ti thiu, c nh t dc dnh v cao.
Bng 2.4. Cc cng thc thit k ti u cng thc vin nang.
Cao Dip c
MgCO3
T dc
n A
T dc
n B
PVP K30
Talc

T1
T2
T3
T4
T5
T6
T7
T8
T9 T10
A% A% A% A% A% A% A% A% A% A%
X1% X1% X1% X1% X1% X2% X2% X2% X2% X2%
Y1% Y2% Y3% Y4% Y5% Y1% Y2% Y3% Y4% Y5%
Y5% Y4% Y3% Y2% Y1% Y5% Y4% Y3% Y2% Y1%
2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

2%
1%

Tin hnh kho st cc c tnh ca khi bt thuc: gc chy, s phn b c

ht, m; o r ca vin nang thnh phm v chn cng thc ti u.

2.2.2.2. Phng php kho st c tnh ca khi bt thuc v o r ca

vin nang:

22

o m ca khi bt thuc: o m bng cn sy m hng ngoi vi cc

thng s nh sau
- Lng cn: khong 3g.
- Nhit sy: 105oC.
- Tc m ti hn: 0,01%/pht.
Kho st s phn b c ht:
Cn 50g bt thuc, cho qua b ry gm cc ry 0,45mm; 0,3mm; 0,2mm, lc
u 5 pht, cn lng bt cn li trn mi ry, v ng phn b c ht.
Xc nh t trng biu kin trc v sau khi g:
Xc nh t trng biu kin trc khi g (t trng th) v t trng biu kin sau
khi g (t trng v) theo cng thc sau:

M
Vb

Vi:

: T trng biu kin ca ht.

M: Khi lng ca ht.
Vb: Th tch biu kin ca ht.
T trng biu kin sau khi g xc nh theo USP30 trn my o t trng biu
kin Pharma test touch.
Xc nh lu tnh ca ht: o gc chy
Cn 50g bt thuc, ht chy lin tc to thnh khi chp v xc nh gc
ngh bit:

tg =
Trong :

h
r

h: chiu cao khi bt.

r: bn knh y ca khi bt.

23

2r
Hnh 2.1. Phng php xc nh gc ngh ca bt, ht thuc.
tan r:
Thc hin theo DVN IV, ph lc 11.4.
2.2.2.3. Quy trnh iu ch vin nang cha cao dip c:
- Trn u cao Dip c vi magnesi carbonat, t dc n A v B bng tay sau
bng my trn, sy khi bt 50oC trong 12 gi.
- Pha t dc dnh: pha PVP K30 nng 7,5% trong cn 50%.
- Trn u khi bt thuc vi t dc dnh, st ht qua ry 1,2mm, tc vng
70 vng/pht, sau sy trong t sy 12 gi 50oC.
- Sa ht qua ry 0,6mm, tc vng 50 vng/pht, thm bt talc, trn hnh
tinh 5 pht.
- ng nang s 0 bng my ng nang th cng (the capsule machine).
2.2.3. Xy dng tiu chun kim nghim cho vin nang cha cao Dip c:
2.2.3.1. Hnh thc cm quan: quan st bng mt thng.
2.2.3.2. ng u khi lng: th theo DVN IV, ph lc 11.3.
2.2.3.3. tan r: th theo DVN IV, ph lc 11.6.
2.2.3.4. m: o m bng cn sy m hng ngoi ging muc 2.2.2.2.
2.2.3.5. nh tnh:
Bng phn ng ha hc:
Cho bt thuc trong nang vo ci chy v nghin thnh bt mn. Cn mt lng
bt thuc tng ng khong 0,5g cao Dip c ha tan trong khong 15ml cn 96%,
un nng nh. Lc ly dch lc chia lm 3 ng nghim lm cc phn ng.
Bng 2.5. nh tnh vin nang cha cao Dip c bng phn ng ha hc.

24

Thuc th
NaOH 1%
FeCl3 1%

Mg + HCl m c

Cch tin hnh

Nh vo ng nghim vi git

Yu cu
Dung dch chuyn sang

NaOH 1%.
Nh vo ng nghim vi git

mu vng sng.
Dung dch chuyn sang

FeCl3 1%.
Cho dch chit vo ng nghim

mu xanh en.

ln cha sn mt t bt Mg kim
loi. Thm t t theo thnh ng

Xut hin mu .

nghim 1 2 ml HCl m c.
Bng sc k lp mng:
Mu chun: cho mt t tinh th quercetin vo ng nghim, thm vi git
methanol ha tan.
Mu th: cn mt lng bt thuc nghin mn tng ng vi khong 0,5g
cao Dip c, ha tan trong 5ml cn 96%, lc thu ly dch lc. Thm vo 5ml dung
dch HCl 10% ri un cch thy thy phn. Lc dch thy phn vi 10ml
chloroform, ly dch thy phn chm sc k.
Bn mng: silicagel F254 (Merck).
H dung mi: Chloroform Ethyl acetat Acid formic (5:4:1).
Pht hin: bng n UV bc sng 254nm, nhng thuc th FeCl3 trong cn.
2.2.3.6. nh lng:
nh lng bng phng php HPLC.
Cc iu kin HPLC thc hin theo cc iu kin ti u kho st, dung dch
chun v th thc hin theo muc 2.2.1.2.
Hm lng quercetin trong mt nang:

X ( g / vin) =

S t Cc 10 P a
Sc m

Trong :
St: din tch nh mu th.

25

Sc: din tch nh mu chun.

Cc: nng cht chun trong dung dch (g/ml).
m: khi lng bt thuc cn (gam).
P: khi lng trung bnh ca vin (gam).
a: Ham lng cao trong vin nang.
Yu cu: hm lng quercetin nm trong gii hn 218,25 g n 266,76 g
tnh theo khi lng trung bnh ca bt thuc trong nang.

Chng 3 - KT QU NGHIN CU

26

3.1. XY DNG V THM NH QUY TRNH NH LNG

QUERCETIN BNG PHNG PHP HPLC:
3.1.1. Kho st cc iu kin sc k ti u:

Hnh 3.1. Sc k 3D mu th bc sng t 250nm 450nm.

Hnh 3.2. Sc k contour mu th.

27

Nhn xt: bc sng 370nm, quercetin c nh cc i hp thu t b nh

hng bi cc tp khc c trong mu th.
Qua thc nghim chng ti xut cc iu kin sc k ti u nh sau:
-

Ct: LiChrospher RP-18 (250 x 4mm, 5m).

Dung mi pha ng: MeOH - dich c acid phosphoric 0,2% (60:40).

Bc sng pht hin: 370nm.

Tc dng: 0,8ml/pht.

Nhit : 25oC.

3.1.2. Kho st quy trnh x l mu cao Dip c v vin nang cha cao Dip c:

1h

2h

3h

1h

2h

3h

UV 254
Thuc th FeCl3
Hnh 3.3. Sc k cc dch thy phn quercetin sau 1h, 2h v 3h.
Nhn xt: qua 3 gi thy phn thy phn ht cc quercetin dng glycosid c
trong mu th.

28

UV 254
Thuc th FeCl3
Hnh 3.4. Sc k mu th trc v sau khi loi tp bng SPE t ch.
Ch thch:
-

1. Dch CHCl3 trc khi qua ct SPE

2. Dch CHCl3 loi tp

3. Dch cha Quercetin

Nhn xt: sau khi qua SPE t ch loi c phn ln cc tp km phn cc
-

v cc tp phn cc trong mu th.

3.1.3. Thm nh quy trnh nh lng quercetin:

3.1.3.1. Xc nh tnh tng thch h thng:

29

Kt qu thc nghim nh gi tnh tng thch h thng ca phng php

HPLC nh lng quercetin c trnh by trong bng 3.1. v bang 3.2.
Bng 3.1. Kt qu kho st tnh tng thch h thng trn 6 ln tim mu chun.
STT

Thi gian

Din tch

H s dung

S a

lu (tR)
7,567
7,467
7,580
7,693
7,727
7,527
7,594
0,098
1,29

nh (S)
29435569
28338432
28285507
28461905
29062188
28337809
28653568,330
479523,498
1,673

lng (k)
1,410
1,417
1,442
1,478
1,440
1,457
1,441
0,025
1,735

l thuyt (N)
629
600
618
628
633
622
622
12
1,929

1
2
3
4
5
6
TB
SD
RSD%

Bng 3.2. Kt qu kho st tnh tng thch h thng trn 6 ln tim mu th.
STT

Thi gian

Din tch

H s dung

lu (tR)
nh (S)
lng (k)
1
7,633
60625893
1,461
2
7,427
58274555
1,400
3
7,400
58195873
1,418
4
7,487
59030548
1,421
5
7,573
59311624
1,427
6
7,647
59127296
1,457
TB
7,528
59094298,170
1,431
SD
0,015
477612,043
0,024
RSD%
0,199
0,808
1,677
3.1.3.2. Thm nh phng php phn tch:
a. Tnh c hiu:

S a
l thuyt (N)
653
660
677
679
675
678
669
14
1,644

30

Hnh 3.5. Sc k mu chun.

Hnh 3.6. Sc k mu th.

Hnh 3.7 Sc k mu trng.

31

Hnh 3.8. Sc k mu dung mi.

Hnh 3.9. Sc k mu dung dich nc acid phosphoric 0,2%.

Hnh 3.10. Kt qu chng ph mu chun, mu dung mi, mu t dc

v mu dung dch nc acid phosphoric 0,2%.

32

Nhn xt: mu trng, mu dung mi, mu nc acid khng c nh hp thu ti

thi gian lu 7,727 pht. Quy trnh t tnh c hiu.
b. Tnh tuyn tnh v min gi tr:
Bng 3.3.Kt qu kho st tnh tuyn tnh ca phng php nh lng bng HPLC.
Mu chun
1
2
3
4
5
6

Nng (g/ml)
50
100
150
200
250
300

Din tch nh
28285507
60085198
90543111
121819257
161937814
191125387

Din tch nh

y = 657733,368x 6137293,733
R2 = 0,9991
y = 657733,368x 6137293,733
R2 = 0,9991

Nng (g/ml)
Hnh 3.11. Biu tnh tuyn tnh ca phng php nh lng bng HPLC.
Nhn xt: R2 = 0,9991 > 0,99 trong khong [50g/ml, 300g/ml]. C s tng
quan r rt gia hp thu v nng trong khong [50g/ml, 300g/ml].
Kho st thng k bng phn mm Ms-Excel vi cng c Regression cho thy:
-

F = 1205,280 > 6,61. Phng trnh hi quy c tnh tng thch.

H s Bo = - 6137293,733, to = -2,033 < t0,05 = 2,571. H s Bo khng c ngha.

H s B = 657733,368, t = 34,717 > t0,05 = 2,571. H s B c ngha.

Phng trnh hi quy c dng: y = 657733,368x

33

c. chnh xc:
Bng 3.4. Kt qu kho st chnh xc ca phng php nh lng bng HPLC.
Mu
1
2
3
4
5

Hm lng (g)

Kt qu thng k
quercetin/1g cao Dip c
928,48
N = 6, f = 5, X = 970,02
925,46
SD = 37,76
990,12
959,73
RSD% = 3,89%
1008,35
P = 95% ( = 0,05)

1007,96

t = 2,57, e = 39,62

Kt lun: RSD% = 3,89% < 5,3%. Quy trnh t chnh xc theo AOAC.
d. ng:
Bng 3.5. Kt qu kho st ng ca phng php nh lng bng HPLC.
Mu

Mc kho

Lng thm

Lng tm

T l phc

T l phc

st

vo (g)
800
800
800
1000
1000
1000
1200
1200
1200

thy (g)
784,293
742,214
774,210
909,080
914,122
892,058
1145,754
1152,425
1164,682

hi (%)
98,037
92,777
96,776
90,908
91,412
89,206
95,479
96,035
97,057

hi TB (%)

1
2
80%
95,863
3
4
5
100%
90,509
6
7
8
120%
96,191
9
TB
94,187
Kt lun: t l hi phc l 94,187% trong khong (90 107%), quy trnh t
ng theo AOAC.
3.2. THIT K CNG THC VIN NANG CHA CAO DIP C:
3.2.1. La chn hm lng cao:
Bng 3.6. Cc cng thc thit k xc nh hm lng cao thch hp.
Cao Dip c
MgCO3
T dc n A

C1
30%
9%
58%

C2
40%
12%
45%

C3
50%
15%
32%

C4
60%
18%
19%

34

PVP K30
Talc

2%
1%

2%
1%

2%
1%

2%
1%

Bng 3.7. m cc cng thc C1, C2, C3, C4.

Ln o 1 Ln o 2 Ln o 3 Trung bnh
C1
3,62%
3,73%
3,80%
3,72%
C2
3,94%
4,04%
4,03%
4,00%
C3
4,57%
4,63%
4,60%
4,60%
C4
5,30%
5,44%
5,59%
5,44%
Nhn xt: Cng thc vi hm lng cao 60% c m > 5%, v vy cng thc
vi hm lng cao 50% c chn.
3.2.2. La chn hm lng t dc ht:
Bng 3.8. Cc cng thc thit k xc nh hm lng t dc ht.
Cao Dip c
MgCO3
T dc n B
PVP K30
Talc

H1
50%
0%
47%
2%
1%

H2
50%
5%
42%
2%
1%

H3
50%
10%
37%
2%
1%

H4
50%
15%
32%
2%
1%

Bng 3.9. m cc cng thc H1, H2, H3, H4.

Ln o 1 Ln o 2 Ln o 3
H1
H2
5,52%
5,38%
5,00%
H3
4,86%
4,75%
4,73%
H4
4,40%
4,52%
4,43%
Ch thch: (-) cc cng thc khng thc hin c.

Trung bnh
5,30%
4,78%
4,45%

Nhn xt: Khi chon hm lng MgCO3 10% th cng thc t m 5%.
V vy, chng ti chn hm lng MgCO3 2 mc 10% v 15% a vo thit k
cng thc.
3.2.3. La chn cng thc ti u cho vin nang cha cao Dip c:
Bng 3.10. Cc cng thc thit k ti u cng thc vin nang.

35

Cng thc
T1
T2
T3
T4
T5
T6
T7
T8
T9
T10

Cao

Magnesi

T dc

T dc

PVP

Dip c
50%
50%
50%
50%
50%
50%
50%
50%
50%
50%

carbonat
10%
10%
10%
10%
10%
15%
15%
15%
15%
15%

n A
27,7%
24,7%
18,5%
12,3%
9,3%
27,7%
24,7%
18,5%
12,3%
9,3%

n B
9,3%
12,3%
18,5%
24,7%
27,7%
9,3%
12,3%
18,5%
24,7%
27,7%

K30
2%
2%
2%
2%
2%
2%
2%
2%
2%
2%

Talc
1%
1%
1%
1%
1%
1%
1%
1%
1%
1%

3.2.3.1. m:
Bng 3.11. m cc cng thc kho st (T1 T10).
Ln 1

T1
T2
T3
T4
T5
T6
T7
T8
T9
4,73% 4,71% 4,80% 4,70% 4,75% 4,52% 4,54% 4,60% 4,53

T10
4,60

Ln 2

%
4,70% 4,68% 4,73% 4,75% 4,80% 4.51% 4,51% 4,70% 4,56

%
4,56

Ln 3

%
4,76% 4,71% 4,72% 4,71% 4,87% 4,47% 4,45% 4,68% 4,56

%
4,67

%
4,75% 4,72% 4,81% 4,50% 4,50% 4,66% 4,55

%
4,61

TB

4,73% 4,7%

Nhn thy cc cng thc u c m <5% , t yu cu v m.

3.2.3.2. Gc chy:
Bng 3.12. Gc chy cc cng thc kho st (T1 T10).
h
r

T1
3,4
4,5

tg

0,756

T2
3,8
4,8

T3
3,5
4,7

T4
3,5
4,5

T5
3,5
4,6

T6
3,4
5

T7
3,1
4,7

T8
3,5
4,5

T9
3,4
4,5

0,792 0,745 0,778 0,761 0,680 0,660 0,778 0,756

T10
3,5
4,6
0,76
1

36

37,1o 38,4o 36,7o 37,9o 37,3o 34,2o 33,4o 37,9o 37,1o 37,3o
Nhn xt: cc cng thc kho st u c gc chy tt, ph hp ng nang.

3.2.3.3. T trng biu kin:

Bng 3.13. T trng biu kin ca cc cng thc kho st (T1 T10).
T1
T2
T3
T4
T5
Vtv (ml)
78
80
78
75
72
Vsv (ml)
67
68
69
66
64
dtv (g/ml) 0,64 0,63 0,64 0,67 0,69
dsv (g/ml) 0,75 0,74 0,72 0,76 0,78
Nhn xt: cc cng thc u c t trng sau

T6
81
70
0,62
0,71
va p

T7
T8
T9
T10
76
74
76
76
67
66
67
67
0,66 0,68 0,66 0,66
0,75 0,76 0,75 0,75
trong khong 0,71 0,78.

T tnh c phn sut nn < 5%.

3.2.3.4. Phn b c ht:

Bng 3.14. Phn b c ht ca cc cng thc kho st (T1 T10).
T1
T2
T3
T4
T5
T6
T7
T8
T9
T10

<0,2 mm
42,80%
34,58%
27,50%
9,98%
7,04%
28,20%
20,74%
13,36%
17,68%
13,02%

0,2 0,3 mm
26,60%
25,66%
29,10%
18,04%
22,20%
30,72%
27,50%
25,06%
30,00%
26,94%

0,3 0,45 mm
23,30%
28,04%
31,72%
45,14%
45,58%
32,56%
40,02%
42,64%
37,50%
44,36%

0,45 0,6 mm
7,30%
11,72%
11,68%
26,84%
21,44%
8,52%
11,74%
18,84%
14,82%
15,68%

37

Hnh 3.12. Biu phn b c ht ca cc cng thc t T1 n T5.

Hnh 3.13. Biu phn b c ht ca cc cng thc t T6 n T10.

T cc biu 3.12. va biu 3.13. cho thy:
- Cc cng thc T1, T2, T3, T6, T7 c t l ht mn tng i ln (> 20%).

38
- Cc cng thc T4, T5, T8, T9, T10 c t l ht mn nh (< 20%) v phn b

c ht ph hp cho ng nang.
3.2.3.5. tan r:
Bng 3.15. tan r cc cng thc kho st (T1 T10).
Cng thc
Thi gian
r (pht)

T1

T2

T3

T4

T5

T6

T7

T8

T9

T10

10

10,5

9,5

8,5

8,5

7,5

7,5

Cc cng thc u c thi gian r < 30 pht, t yu cu v tan r ca vin

nang theo DVN IV. Trong cng thc T9 v T10 c thi gian r ngn nht.
T cc kt qu trn chng ti chn cng thc T9 lm cng thc ti u cho vin
nang cha cao Dip c.

3.3. KIM NGHIM VIN NANG CHA CAO DIP C:

3.3.1. Hnh thc cm quan:
Vin nang cng s 0, u xanh m, u xanh nht. Bt thuc trong nang mu
nu en, v ng hi ngt, mi c trng.
3.3.2. ng u khi lng:
Bng 3.16. Kt qu kim nghim ng u v khi lng.
STT
1
2
3
4
5
6
7

Khi lng

Khi lng

Khi lng

% So vi

% Chnh lch

nang thuc
0,5855
0,5976
0,5732
0,5901
0,5771
0,5651
0,5972

nang rng
0,0969
0,0976
0,0947
0,097
0,0956
0,089
0,0959

bt thuc
0,4886
0,5000
0,4785
0,4931
0,4815
0,4761
0,5013

KLTB
98,74
101,04
96,69
99,65
97,30
96,21
101,30

so vi KLTB
1,26
1,04
3,31
0,35
2,70
3,79
1,30

39

8
0,5812
0,0972
0,4840
9
0,5991
0,0989
0,5002
10
0,6050
0,0963
0,5087
11
0,5942
0,0954
0,4988
12
0,5723
0,0922
0,4801
13
0,6129
0,0992
0,5137
14
0,6053
0,096
0,5093
15
0,6135
0,0952
0,5183
16
0,5876
0,0997
0,4879
17
0,6112
0,0967
0,5145
18
0,5783
0,0947
0,4836
19
0,5933
0,0901
0,5032
20
0,5699
0,0942
0,4757
KLTB = 0,4949
3.3.3. tan r: Thi gian r: 7,5 pht t.

97,81
101,08
102,80
100,80
97,02
103,81
102,92
104,74
98,59
103,97
97,73
101,69
96,13

Bang 3.17. Kt qua o ra.

Ln 1
Ln 2
Ln 3
TB
7,5 phut
7,5 phut
7,5 phut
7,5 phut
3.3.4. m: Kt qu trung bnh ca 3 ln o l 4,52% t.
Bang 3.18. Kt qua o m.
Ln 1
4,45%

Ln 2
4,6%

Ln 3
4,51%

TB
4,52%

3.3.5. nh tnh:
3.3.5.1. Bng phn ng ha hc:
Bng 3.19. Kt qu nh tnh vin nang cha cao Dip c.
Thuc th
NaOH 10%
FeCl3 5%/ cn 96%
Mg + HCl m c
3.3.5.2. Bng SKLM:
Kt qu:

Kt qu
Mu vng sng (+)
Mu xang en (+)
Mu (+)

2,19
1,08
2,80
0,80
2,98
3,81
2,92
4,74
1,41
3,97
2,27
1,69
3,87

40

UV 254
Thuc th FeCl3
Hnhlng:
3.14. Kt qu nh tnh bng SKLM vin nang cha cao Dip c.
3.3.6. nh
Kt qu nh lng vin nang cha cao Dip c: p dng cng thc muc 2.2.3.6.
Sc = 60085198; St = 62727371; Cc = 100 (g/ml); m = 1,0334g; P = 0,4959g
Hm lng trong 1 vin: 249,98 g t.
3.4. XY DNG TIU CHUN VIN NANG CHA CAO DIP C:
3.4.1. Tiu chun k thut:
1. YU CU K THUT:
1.1. Cng thc pha ch cho mt vin nang:
Cao Dip c

250mg

T dc va

1 vin

1.2. Nguyn liu:

Cao Dip c
T dc A
T dc B
Magnesi carbonate
Talc
Povidon K30
Cn 96%
1.3. Cht lng thnh phm:

t tiu chun c s
t tiu chun
t tiu chun
t tiu chun c s
t tiu chun USP 27
t tiu chun USP 27
t tiu chun DVN IV

41

1.3.1. Tnh cht: Vin nang cng s 0, u xanh m, u xanh nht. Bt thuc
trong nang mu nu en, v ng hi ngt, mi c trng.
1.3.2. ng u khi lng: 7,5% so vi khi lng trung bnh ca bt thuc
trong nang.
1.3.3. tan r: khng qu 30 pht.
1.3.4. m: khng qu 5%.
1.3.5. nh tnh: Phi c phn ng c trng ca Dip c.
1.3.6. nh lng: mi vin phi cha quercetin trong gii hn 218,25 g n
266,76 g tnh theo khi lng trung bnh ca bt thuc trong nang.
2. PHNG PHP TH:
2.1. Tnh cht: kim tra bng cm quan ch phm phi t cc yu cu nu.
2.2. ng u khi lng: th theo DVN IV, ph lc 11.3.
2.3. tan r: th theo DVN IV, ph lc 11.6.
2.4. m: o m bng cn sy m hng ngoi vi cc thng s nh sau:
- Lng cn: khong 3g.
- Nhit sy: 105oC.
- Tc m ti hn: 0,01%/pht.
2.5. nh tnh:
A. Bng phn ng ha hc:
Cho bt thuc trong nang vo ci chy v nghin thnh bt mn. Cn mt lng bt
thuc tng ng khong 0,5g cao Dip c ha tan trong khong 15ml cn 96%, un
nng nh. Lc ly dch lc chia lm 3 ng nghim lm cc phn ng.
Nh vo ng nghim th nht vi git Natri hydroxyd 1% (TT). Dung dch chuyn
sang mu vng sng.
Nh vo ng nghim th hai vi git St (III) clorid 1% (TT). Dung dch chuyn
sang mu xanh en.
Cho dch chit vo ng nghim th 3 cha sn mt t bt Magnesi kim loi.
Thm t t theo thnh ng nghim 1 2 ml dung dch HCl m c. Dung dch
xut hin mu .

42

B. Bng sc k lp mng:
Bn mng Silicagel F254 (Merck).
Dung mi khai trin: Chloroform Ethyl acetat Acid formic (5:4:1).
Dung dch chun: cho mt t tinh th quercetin vo ng nghim, thm vi git
methanol ha tan.
Dung dch th: cn mt lng bt thuc nghin mn tng ng vi khong 0,5g
cao Dip c, ha tan trong 5ml cn 96%, lc thu ly dch lc. Thm vo 5ml dung
dch HCl 10% (TT) ri un cch thy thy phn. Lc dch thy phn vi 10ml
chloroform, ly dch chloroform chm sc k.
Cch tin hnh: chm ring bit ln bn mng 20l mi dung dch. Trin khai sc
k cho n khi dung mi i c khong 10cm. Lm kh bn mng, kim tra di
nh sng n t ngoi bc sng 254nm v bng dung dch St (III) clorid 1%/
cn 96% (TT). Trn sc k ca dung dch th phi c vt trng vi sc k ca
dung dch chun.
2.6. nh lng:
Tin hnh bng phng php sc k lng.
Pha ng: Methanol dung dch nc acid phosphoric 0,2% (60:40).
Dung dch chun: cn chnh xc khong 50mg quercetin chun vo trong bnh nh
mc 50ml, ha tan v pha long bng methanol (TT) n nh mc, trn u. Pha
long 5ml dung dch thu c thnh 50ml vi methanol (TT), trn u. Lc qua
mng lc 0,45m.
Dung dch th:
Cn 20 nang, tnh khi lng trung bnh ca bt thuc trong nang, nghin thnh bt
mn. Cn lng bt thuc tng ng vi 1g cao Dip c, ha tan trong 10ml cn
25% (TT), lc thu ly dch lc, ra cn vi khong 2ml cn 25% (TT), thm vo
20ml chloroform, lc u 5 pht, chuyn vo bnh lng gn, rt b dch chloroform.
Dch nc cn li ha tan vi 10ml dung dch HCl 10%/cn 25% un hi lu. Dch
thy phn sau lc vi 15ml chloroform trong 5 pht, chuyn vo bnh lng gn
yn 60 pht, rt ly dch chloroform. Lp li n khi chit kit quercetin. Gp

43

dch chloroform lc vi 20ml nc. Rt dch chloroform lc qua natri sulfat khan
sau em c n dch chloroform m c.
Dch Chloroform m c cho qua ct SPE t ch (1,0g silica kch thc ht 40m
63m, chiu di ct 70mm, ng knh 12mm) loi tp. Cho 20ml chloroform
qua ct SPE loi tp phn cc, tip theo cho 40ml hn hp CHCl3: EtOAc 6:4
ly phn on cha quercetin. C phn on cha quercetin n cn.
Ha tan cn trong methanol ri cho vo bnh nh mc 10ml, b sung th tch,
lc s b. Sau lc qua mng lc 0,45m.
iu kin sc k:
Ct thp khng g (25cm x 4,0mm) c nhi pha tnh C (5m).
Nhit ct: 25oC.
Detector DAD, bc sng pht hin 370nm.
Tc dng: 0,8ml/pht.
Th tch tim: 20l.
Cch tin hnh:
Kim tra kh nng thch hp ca h thng sc k: Tin hnh sc k i vi dung
dch chun. Php th ch c gi tr khi lch chun tng i ca din tch peak
quercetin trong 6 ln tim lp li nh hn 2,0%.
Tin hnh sc k ln lt i vi dung dch chun v dung dch th.
Tnh hm lng quercetin c trong mt vin nang da vo din tch peak trn sc
k thu c ca dung dch th, dung dch chun v hm lng quercetin c
trong mu chun.
3. NG GI GHI NHN - BO QUN:
ng gi: ng trong chai nha c cht ht m, mi chai 100 vin hoc p v bm,
mi v 10 vin.
Nhn: ng quy ch.
Bo qun: Ni kh mt, trnh nh sng, nhit khng qu 30oC.

44

3.4.2. Kt qu kim nghim:

Bng 3.20. Kt qu kim nghim vin nang Dip c.
Ch tiu
Tnh cht

Tiu chun
Kt qu
Vin nang cng s 0, u xanh ng
m, u xanh nht. Bt thuc
trong nang mu nu en, v ng
hi ngt, mi c trng.
Khi lng trung bnh 7,5%

t (0,4949

Khng qu 30 pht
Khng qu 5%

4,79%)
t (7,5 pht)
t (4,52%)

Dd NaOH 1%

Mu vng sng

ng

Dd FeCl3 1%

Mu xanh en

ng

Mg/HCl m c

Mu

ng

ng u khi lng
tan r
m
nh tnh
A. Bng phn ng ha hc

B. Bng SKLM

Dung dch th phi cho mt vt ng

tng ng v mu sc v Rf vi

nh lng

dung dch chun.

Mi vin phi cha quercetin trong t
gii hn 218,25 g n 266,76 g (249,98g/vin)
tnh theo khi lng trung bnh ca

45

bt thuc trong nang.

Kt lun: Vin nang Dip c t tiu chun c s.

Chng 4 - BN LUN
4.1. V XY DNG V THM NH QUY TRNH NH LNG
QUERCETIN TRONG CAO DIP C V VIN NANG CHA CAO DIP C:

Dip c cha phn ln cc flavonoid co aglycon l quercetin. V vy, dng

quercetin nh l cht nh du kim sot cht lng cao Dip c v vin nang
cha cao Dip c l ph hp.
Mu cao Dip c v mu vin nang cha cao Dip c c x l m bo
qu trnh thy phn cc flavonoid c xy ra hon ton, qu trnh loi tp c
trit , m bo an ton cho h thng HPLC.
Vi kt qu thu c sau khi thm nh, cc yu cu v tnh tng thch h
thng, tnh c hiu, ng, chnh xc, tnh tuyn tnh v min gi tr ca
phng php HPLC cho cao Dip c v vin nang cha cao Dip c cho thy
phng php p ng quy trnh nh lng v khng b nh hng bi s hin din
ca cht khc nh t dc, dung mi ha tan mu,...
4.2. V NGHIN CU XY DNG CNG THC CHO VIN NANG
CHA CAO DIP C:
Bo ch vin nang thng khng qu phc tp nh vin nn, thnh phn khi
thuc tng i n gin, khng phi dng nhiu loi t dc nh vin nn. Mt
khc, bo ch vin nn t dc liu khng cho cm quan tt. Vin sau khi nn phi
c bao ng hoc bao phim, i hi k thut v thit b phc tp. V vy, ti
chn dng bo ch l vin nang.
bo ch vin nang th yu t quan trng nht ca khi bt thuc l hm m
v kh nng ht m v chng c th lm h v nang. Ngoi ra, cn phi chn t

46

dc ph hp sao cho lng t dc s dng l t nht. Bng cc thit k nghin

cu n gin ca Gauss Geidell, ti chn c t dc ht m l MgCO 3
vi hm lng l 15%, t dc n ph hp, hm lng cao trong khi bt thuc l
50%.
m bo chc chn vin nang sau khi bo ch t ng u khi lng
th khi bt thuc bo ch c phi c t l ht mn nh, phn b kch thc ht
ph hp v trn chy tt. thc hin iu , ti s dng povidon K30
lm t dc dnh gim t l bt mn, s dng talc khi bt thuc trn chy tt
ng thi thit k la chn cng thc c phn b c ht ph hp.
V tan r, vin nang bo ch c c tan r tt. Thi gian r l 7,5 pht,
thp hn nhiu so vi tiu chun tan r ca vin nang theo DVN IV l khng
qu 30 pht.
4.3. V XY DNG TIU CHUN KIM NGHIM CHO VIN NANG
CHA CAO DIP C:
Vic xy dng tiu chun kim nghm vin nang cha cao Dip c c da
trn tiu chun kim nghim thuc vin nang theo DVN IV bao gm cc ch tiu:
Hnh thc cm quan, ng u khi lng, tan ra, m, nh tnh, nh
lng. Vin nang bo ch c t cc ch tiu trn.
Tuy nhin, tiu chun kim nghim cn b sung thm ch tiu nhim khun
v c tnh bt thng m bo rng qu trnh chuyn dng bo ch khng lm
pht sinh c tnh bt thng cng nh s xm nhp ca cc vi khun, nm mc
gy bnh.

47

Chng 5 - KT LUN V KIN NGH

5.1. KT LUN:
ti xy dng v thm nh quy trnh nh lng quercetin trong cao Dip
c v vin nang cha cao Dip c t cc yu cu v tnh tng thch h thng, tnh
c hiu, tnh tuyn tnh, chnh xc v ng.
V nghin cu xy dng cng thc vin nang cha cao Dip c: xy dng
c cng thc v bo ch thnh cng vin nang cha cao Dip c t cc ch tiu
ra.
Cng thc cho mt vin nang nh sau:
Cao Dip c
250mg
T dc A
61,5mg
T dc B
123,5mg
Magnesi carbonate
75mg
Talc
5mg
Povidon K30
10mg
V xy dng tiu chun kim nghim cho vin nang cha cao Dip c: xy
dng c bn d tho tiu chun kim nghim da vo cc tiu chun kim
nghim vin nang theo DVN IV. Cac tiu chun kim nghim bao gm:
1. Tnh cht: Vin nang cng s 0, u xanh m, u xanh nht. Bt thuc
trong nang mu nu en, v ng hi ngt, mi c trng.
2. ng u khi lng: 7,5% so vi khi lng trung bnh ca bt thuc
trong nang.
3. tan r: khng qu 30 pht.
4. m: khng qu 5%.
5. nh tnh: Phi c phn ng c trng ca Dip c.

48

6. nh lng: mi vin phi cha quercetin trong gii hn 218,25 g n

266,76 g tnh theo khi lng trung bnh ca bt thuc trong nang.

5.2. KIN NGH:

a thm ch tiu c tnh bt thng v ch tiu nhim khun vo tiu
chun kim nghim cho vin nang cha cao Dip c.
Nghin cu s n nh ca ch phm.
Tin hnh sn xut mt s m quy m pilot thm nh li cc thng s
cng ngh trc khi trin khai sn xut ln.

49

TI LIU THAM KHO

Ting vit:
1. B mn Bo ch - Trng i Hc Dc H Ni (2008), Bo ch v sinh
dc hc cc dng thuc tp 2, NXB Y Hc, H Ni.
2. B mn Bo ch - Trng i Hc Y Dc TP.HCM (2007), Bo ch hc
tp 2, NXB Y Hc chi nhnh TP. H Ch Minh, TP. H Ch Minh.
3. B mn Ha phn tch, Kim nghim, c cht - Trng i Hc Y Dc
Cn Th (2010), gio trnh Kim nghim thuc, ti liu lu hnh ni b, Cn
Th.
4. B mn Ha phn tch, Kim nghim, c cht - Trng i Hc Y Dc
Cn Th (2009), Ha phn tch 2, Ti liu lu hnh ni b, Cn Th.
5. B Y T (2009), Dc in Vit Nam IV, NXB Y Hc, H Ni.
6. Huy Bch, ng Quang Chung, Bi Xun Chng, (2004), Cy thuc
v ng vt lm thuc Vit Nam, NXB Khoa hc v K thut, H Ni, tr.
321-323.
7. V Th Kim Chi (2008), Nghin cu bo ch vin nang Linh chi - ng
quy, lun vn tt nghip Dc s i hc, Trng i Hc Y Dc TP.HCM,
TP. H Ch Minh.
8. Nguyn Th Chung (2007), ng dng ti u ha thng k trong nghin cu
pht trin dc phm, Ti liu lu hnh ni b, TP. H Ch Minh.
9. Nguyn Minh c (2006), Sc k lng hiu nng cao v mt s ng dng
vo nghin cu, kim nghim dc phm, dc liu v hp cht t nhin,
NXB Y Hc chi nhnh TP. H Ch Minh, TP. H Ch Minh.

50

10. Trn Th Vit Hoa, L Thi Kim Oanh (2008), Phn lp v xc nh cu trc
mt s hp cht t cy Dip C (Houttuynia cordata Thumb.) ca Vit
nam, Tp ch pht trin KH&CN, 11 (07), tr. 73-77.
11. Hong Ngc Hng, V Chu Hng (2006), T dc v cht ph gia dng
trong dc phm, m phm v thc phm, NXB Y Hc, H Ni.
12. Hong Thanh Hng, Trn Qunh Hoa, H Vit Bo, Nguyn Danh Thc
(2002), Gp phn nghin cu thnh phn flavonoid chit xut t l cy Dip
c - Houttuynia cordata Thunb. ca Vit Nam, Tp ch dc hc, (9), 1315.
13. Tt Li (2000), Nhng cy thuc v v thuc Vit Nam, NXB Y hc, H
Ni, tr. 40-41.
14. Nguyn Nht Thnh (2001), Nghin cu k thut iu ch vin nang i b
m t bi thuc cng tn, lun vn tt nghip thc s dc hc, Trng i
Hc Y Dc TP.HCM, TP. H Ch Minh.
15. Nguyn ng Thoi, Trn Ngc Dn, Th Minh Thun, Hong Minh
Chu, Nguyn Thin Hi, Nguyn c Tun (2009), Xy dng quy trnh
nh lng flavonoid ton phn trong cao Bch qu, vin bao phim O.P.Can
v quercetin trong huyt tng ngi bng phng php HPLC, Y hc
TPHCM-Chuyn Dc, 13 (1), tr.78-83.
16. Trng i hc Dc H Ni (2002), Bi ging dc liu. NXB Y hc, H
Ni, tr. 205-207.

Ting Anh
17. Chang Jung - San, Chiang Lien - Chai, Chen Chi - Chain, Liu Li - The,
Wang Kuo - Chih, Lin Chun - Ching (2001), Antileukemic activity of
Bidens pilosa L. var. minor (Blume) Sherff and Houttuynia cordata Thunb,
The American journal of Chinese medicine, 29 (2), pp. 303-312.

51

18. Cho E. J., Yokozawa T., Rhyu D. Y., Kim S. C., Shibahara N., and Park J. C.
(2003), Study on the inhibitory effects on korean medicinal plants and their
main

compounds

on

the

1,1-diphenyl-2-picrylhydrazyl

radical,

Phytomedicine, 10 (6-7), pp. 544-551.

19. Cho E. J., Yokozawa T., Rhyu D. Y., Kim S. C., Shibahara N., and Park J. C.
(2003), The inhibitory effects of 12 medical plants and their compoment
compound no lipid peroxidation , Am. J. Chin. Med, 31 (6), pp. 907-917.
20. Corlett J. L., Clegg M. S., Keen C. L., Grivetti L. E. (2002), Mineral content
of culinary and medicinal plants cultivated by Hmong refugees living in
Sacramento, Califonia, Int. J. Food Sci. Nutr., 53 (2), pp. 117-128.
21. Hayashi, Kyoko; Kamiya, Mioko; Hayashi, Toshimitsu (1995), Virucidal
Effects of the Steam Distillate from Houttuynia cordata and its Components
on HSV-1, Influenza Virus, and HIV, Planta Med, 61(03), pp. 237-241.
22. In Sik Kim, Joo-Hwan Kim, Jin Sook Kim, Chi-Young Yun, Dong-Hee Kim
and Ji-Sook Lee (2007), The inhibitory effect of Houttuynia cordata extract
on stem cell factor-induced HMC-1 cell migration, Journal of
Ethnopharmacology, 112 (1), pp. 90-95.
23. Kim SK, Ryu SY, No J, Choi SU, Kim YS (2001), Cytotoxic alkaloids from
Houttuynia cordata, Korea Research Institute of Chemical Technology,
Taejeon, Arch Pharm Res, 24(6), pp. 518-21.
24. Ladislav Kokoska v Vojtech Rada (2001), Antibacterial Activity of
Houttuynin Sodium Bisulphate (HSB), Czech J. Food Sci, 19 (1), pp. 37-40.
25. Li C, Zhao Y, Liang C, An H (2001)., Observations of the curative effect
with various liquid for post operative irrigation of ESS of treating chronic
sinusitis and nasal polyps, Lin Chuang Er Bi Yan Hou Ke Za Zhi, 15(2), pp.
53-54.

52

26. Lien-Chai Chiang, Jung-San Chang, Chi-Chain Chen, Lean-Teik Ng, ChunChing Lin (2003), Anti-Herpes Simplex Virus Activity of Bidens pilosa and
Houttuynia cordata, The American Journal of Chinese Medicine, 31 (3), pp.
355-362.
27. Meng J., Dong X. P., Zhou Y. S., Jiang Z. H., Leung S. Y., Zhao Z. Z.
(2006), study on chemical constituents of flavonoid in fresh herb of
Houttuynia cordata, Zhongguo Zhong Yao Za Zhi, 32 (16), pp. 1335-1337.
28. Ng L. T., Yen F. L., Liao C. W., Lin C. C. (2007), Protective effect of
Houttuynia cordata extract on bleomycin-induced pulmonary fibrosis in
rate, Am. J. Chin. Med., 35 (3), pp. 465-475.
29. S. H. Hu v A. F. Du (1997), Treatment of Bovine Mastitis with Houttuynin
Sodium Bisulphate, Journal of Veterinary Medicine Series B, 44, pp. 365370.
30. Tutupalli L. V., Chaubal M. G. (1975), Saururaceae. V. Composition of
essential oil from foliage of Houtuynia cordata and chemo systematics of
Saururaceae, Lioydia, 38 (2), pp. 92-96.

Bi bo:
31. B Ti nguyn v Mi trng Vit Nam (2010), Cy dc liu cn c
quan tm bo tn, http://www.monre.gov.vn ngy 15/06/2010.
32. Vin Khoa hc nghin cu Nhn ti Nhn lc (2009), Sn xut thuc t
ngun gc thin nhin: i tt n u cch no?, http://nhantainhanluc.com/
ngy 19/06/2009.

PH LC 1
TIU CHUN QUERCETIN CHUN

PH LC 2
TIU CHUN CAO DIP C

PH LC 3
KT QU 6 MU THM NH CHNH XC
Mu
1
2
3
4
5
6
Trung bnh

Lng cn
1.0428
1.0132
1.0147
1.0253
1.0102
1.0036

Din tch nh
58175756
56340463
60366326
59124310
61205139
60781545
970.02

Nng
96.82
93.77
100.47
98.40
101.86
101.16

Hm lng
928.48
925.46
990.12
959.73
1008.35
1007.96

PH LC 4
KT QU PHN TCH PHNG TRNH HI QUY
SUMMARY
OUTPUT

Nng
(g/ml)
50
100
150
200
250
300

Regression Statistics
0.998757
Multiple R
795
0.997517
R Square
132
Adjusted R
0.996689
Square
51
Standard
3037521.
Error
572
Observations
5

Din tch
nh
28285507
60085198
90543111
121819257
161937814
191125387

ANOVA
df
Regression

Residual

Total

SS
1.11206E
+16
2.76796E
+13
1.11482E
+16

Significanc
MS
F
eF
1.11206E+ 1205.280 5.25464E16
226
05
9.22654E+
12

5
Coefficien
ts
Intercept
50

-8287879
666950.1
62

Standard
Error

t Stat

4075262.8
31
19210.973
22

2.033704
167
34.71714
599

P-value

Lower
95%

0.134861
715
5.25464E05

21257184.
14
605812.27
13

Upper
95%
4681426.
139
728088.0
527

Lower
95.0%
21257184.
14
605812.27
13

Upper
95.0%
4681426.
139
728088.0
527

PH LC 5
KT QU THM NH NG
Mc
kho
st

Lng
cn
1.032
1.069

80%

1.079
1.052
1.095

100%

1.044
1.073
1.049

120%

1.032

Lng
c sn
1,001.0
61
1,036.9
51
1,046.6
52
1,020.4
61
1,062.1
72
1,012.7
01
1,040.8
31
1,017.5
51
1,001.0
61

Lng
thm
vo
800
800
800
1000
1000
1000
1200
1200
1200

Din tch
nh
1087452
46
1083682
84
1109079
77
1175276
13
1203753
23
1160181
34
1331841
01
1321724
26
1319145
54

Nng
o

Lng
tng cng
tm c

178.535

1785.35

177.917

1779.17

182.086

1820.86

192.954

1929.54

197.629

1976.29

190.476

1904.76

218.659

2186.59

216.998

2169.98

216.574

2165.74

Lng
tm thy
784.29
3
742.21
4
774.21
0
909.08
0
914.12
2
892.05
8
1,145.7
54
1,152.4
25
1,164.6
82

T l hi
phc

T l hi
phc trung
bnh

98.037
92.777

95.863

96.776
90.908
91.412

90.509

89.206
95.479
96.035

96.191

97.057
94.187