đề tài: Trich Ly Va Khao Sat Tinh Dau Bac Ha

II HHCC CCNN TTHH

KKHHOOAA KKHHOOAA HHCC TT NNHHIINN BB MMNN HHAA HHCC

--------

TN LONG DY

LY TRCH V KHO ST THNH PHN

HA HC CA TINH DU BC H

(MENTHA ARVENSIS)

LUN VN TT NGHIP I HC

Ngnh: Ha hc

MSSV: 2096740

GIO VIN HNG DN

TS. NGUYN TRNG TUN

ThS. PHM QUC NHIN

Cn Th, 6/2013

II HHCC CCNN TTHH

KKHHOOAA KKHHOOAA HHCC TT NNHHIINN BB MMNN HHAA HHCC

--------

TN LONG DY

LY TRCH V KHO ST THNH PHN

HA HC CA TINH DU BC H

(MENTHA ARVENSIS)

LUN VN TT NGHIP I HC

Ngnh: Ha hc

MSSV: 2096740

Cn Th, 6/2013

i

TRNG I HC CN TH

KHOA KHOA HC T NHIN

Nm hc 2012-2013

ti:

LLYY TTRRCCHH VV KKHHOO SSTT TTHHNNHH PPHHNN HHAA HHCC

CCAA TTIINNHH DDUU BBCC HH ((MMEENNTTHHAA AARRVVEENNSSIISS LL..))

LI CAM OAN ......................................................................................................................................

......................................................................................................................................

......................................................................................................................................

......................................................................................................................................

Cn Th, ngy thng nm 2013

Lun vn tt nghip i hc ngnh: Ha Hc

bo v v c duyt

Hiu trng..

Trng khoa..

Trng b mn Cn b hng dn

Ts. Nguyn Trng Tun Ths Phm Quc Nhin

ii

TRNG I HC CN TH CNG HA X HI CH NGHA VIT NAM

KHOA KHOA HC T NHIN c lp T do Hnh phc

B MN HA HC ------------

Cn Th, ngythng...nm 2013

PHIU NG K TI LUN VN TT NGHIP Nm hc: 2012 2013

1. Cn b hng dn: TS. Nguyn Trng Tun v ThS. Phm Quc Nhin

2. Tn ti: Ly trch v kho st thnh phn ha hc ca tinh du Bc h

(Mentha arvensis L.)

3. a im, thi gian thc hin

a im:

Phng th nghim Ha Sinh - B mn Ha hc, Khoa khoa hc t nhin,

Trng i hc Cn Th.

Thi gian: 12/2012 05/2013

4. S lng sinh vin thc hin: 01

5. H v tn sinh vin: Tn Long Dy MSSV: 2096740

Lp: C nhn Ha hc Kha: K35

6. Mc tiu ca ti:

+ Kho st iu kin ti u khi ly trch tinh du Bc h bng phng

php chng ct li cun hi nc .

+ Xc nh cc ch s ha l ca tinh du Bc h.

+ Xc nh thnh phn tinh du Bc h bng GC MS

7. Cc ni dung chnh: ti gm cc phn

+ Phn 1: Tng quan

+ Phn 2: Thc nghim

+ Phn 3: Kt qu v tho lun

iii

+ Phn 4: Kt lun v kin ngh

8. Cc yu cu h tr cho vic thc hin ti: Thit b, ha cht, kinh ph v

mt s dng c cn thit khc.

9. Kinh ph d tr cho vic thc hin ti: 5.000.000 ng

Sinh vin thc hin

(K tn v ghi r h tn)

Tn Long Dy

KIN CA B MN CN B HNG DN

.... TS. Nguyn Trng Tun ThS. Phm Quc Nhin

KIN CA HI NG LV V TLTN

iv



B MN HA HC ------------


NHN XT V NH GI CA CN B HNG DN 1. Cn b hng dn: TS. Nguyn Trng Tun v ThS. Phm Quc Nhin

2. Tn ti: Ly trch v kho st thnh phn ha hc ca tinh du Bc h (Mentha

arvensis L.)

3. Sinh vin thc hin: Tn Long Dy MSSV: 2096740

Lp C nhn Ha hc

4. Ni dung nhn xt:

a. Nhn xt v hnh thc ca lun vn tt nghip:..........................................

..................................................................................................................................

..................................................................................................................................

b. Nhn xt v ni dung ca lun vn tt nghip ( ngh ghi chi tit v y

): ............................................................................................................

..................................................................................................................................

..................................................................................................................................

Nhng vn cn hn ch: ..................................................................

..................................................................................................................................

..................................................................................................................................

c. Nhn xt i vi sinh vin thc hin ti: ...............................................

..................................................................................................................................

..................................................................................................................................

d. ngh v im: .......................................................................................

..................................................................................................................................

..................................................................................................................................

Cn th, ngy thng nm 2013

Cn b hng dn

v



B MN HA HC ------------


NHN XT V NH GI CA CN B PHN BIN 1. Cn b hng dn: TS. Nguyn Trng Tun v ThS. Phm Quc Nhin

2. Tn ti: Ly trch v kho st thnh phn ha hc ca tinh du Bc h (Mentha

arvensis L.)

3. Sinh vin thc hin: Tn Long Dy MSSV: 2096740

Lp C nhn Ha hc

4. Ni dung nhn xt:

a. Nhn xt v hnh thc ca lun vn tt nghip: .........................................

..................................................................................................................................

..................................................................................................................................

b. Nhn xt v ni dung ca lun vn tt nghip ( ngh ghi chi tit v y

): .................................................................................................................

..................................................................................................................................

Nhng vn cn hn ch: ..................................................................

..................................................................................................................................

..................................................................................................................................

c. Nhn xt i vi sinh vin thc hin ti: ...............................................

..................................................................................................................................

..................................................................................................................................

d. ngh v im: .......................................................................................

..................................................................................................................................

..................................................................................................................................


Cn b phn bin

LI CM N

vi

LI CM N ------------

c ngi trn ging ng ca trng i hc Cn Th hc tp trong sut bn

nm qua l nim vinh d i vi em. Tng bui hc, tng bi ging, tng kinh nghim

m cc thy, cc c truyn t li lun l ngun kin thc qu bu cho em. Nhng

kin thc y gi y tr thnh kinh nghim v cng qu gi gip em thc hin tt

lun vn tt nghip, cng nh l nn tng vng chc cho em bc i nhng bc trn

ng i sau ny. Em xin gi li cm n su sc n:

TS. Nguyn Trng Tun v ThS. Phm Quc Nhin lun quan tm hng dn

v to mi iu kin thun li em c th hon thnh tt lun vn ca mnh.

Qu Thy C ca trng i hc Cn Th, nhng ngi truyn t cho em

nhng kin thc qu bu v b ch lm hnh trang vo i.

Qu Thy C B mn Ha hc v B mn Sinh hc Khoa Khoa hc T nhin,

trng i hc Cn Th to mi iu kin v tn tnh gip em hon thnh tt

lun vn.

C s ging cy trng Bnh Chu (24, in Bin Ph, qun Bnh Tn, TP HCM)

nhit tnh gip em c c ngun nguyn liu tt nht em hon thnh qu

trnh thc nghim.

Cm n cc anh ch v cc bn phng th nghim Ha sinh gip v cho li

khuyn qu bu trong qu trnh em thc hin lun vn.

c bit em xin gi li cm n n gia nh, ngi thn v cc bn trong lp C

nhn Ha K35. Nhng ngi lun bn em, ng vin, ng h v gip em c v

vt cht ln tinh thn em c th hon thnh tt lun vn ca mnh.

Em xin chn thnh cm n tt c mi ngi!


K tn

Tn Long Dy

MC LC

vii

MC LC

------------ LI CAM OAN.........................................................................................................i PHIU NG K TI LUN VN TT NGHIP............................................ii NHN XT V NH GI CA CN B HNG DN....................................iv NHN XT V NH GI CA CN B PHN BIN........................................v LI CM N ............................................................................................................vi MC LC.................................................................................................................vii PH LC HNH ........................................................................................................ix PH LC BNG........................................................................................................x DANH SCH CC T VIT TT ...........................................................................xi LI M U ...........................................................................................................xii CHNG I .................................................................................................................1 Tng quan...................................................................................................................1

I.1 GII THIU H LAMIACEAE .......................................................................1 I.1.1 Phn loi hc[1,2]..........................................................................................2 I.1.2 M t[1,2] .....................................................................................................3 I.1.3 Phn b v thu hi[1,2]..................................................................................3 I.1.4 Cng dng ca Bc h ................................................................................4

I.2 TINH DU........................................................................................................5 I.2.1 Khi qut v tinh du ..................................................................................5 I.2.2 Qu trnh tch ly[3,4,5] .................................................................................5 I.2.3 Tinh du Bc h[3] .....................................................................................6

I.3 TNH HNH NGHIN CU TINH DU BC H ..........................................7 I.3.1 Kho st hot tnh khng vi sinh vt ca tinh du Bc h Mentha piperita L.[13] ......................................................................................................7 I.3.2 Thnh phn tinh du ca Bc h Mentha arvensis L. v Bc h Mentha piperita L. nhng giai on sinh trng khc nhau trong vng Kumaon thuc pha ty Himalaya. ...............................................................................................9 I.3.3 Tch v tng hp menthol..........................................................................12

I.4 MT S PHNG PHP LY TRCH TINH DU BC H........................12 I.4.1 Phng php c hc[5,6].............................................................................13 I.4.2 Phng php dng dung mi ha tan[5,6] ...................................................13 I.4.3 Phng php chng ct li cun hi nc[4,5,6]..........................................14 I.4.4 Phng php chit Soxhlet[5].....................................................................15

CHNG II: THC NGHIM.................................................................................16 II.1 A IM, THI GIAN, THIT B, NGUYN LIU V HA CHT......16

II.1.1 a im v thi gian ..............................................................................16 II.1.2 Thit b, nguyn liu, dng c v ha cht ...............................................16

II.2 PHNG PHP NGHIN CU...................................................................17 II.2.1 X l nguyn liu ....................................................................................18 II.2.2 Ly trch tinh du bng phng php li cun hi nc[7] .........................18 II.2.3 Xc nh mt s ch tiu ha l ca tinh du Bc h .............................19

MC LC

viii

II.2.4 Kho st cc yu t nh hng n hm lng v thnh phn tinh du Bc h ...............................................................................................................22 II.2.5 Xc nh thnh phn ha hc chnh ca tinh du Bc h bng phng php sc k ghp khi ph (GC MS) [4,6,7,8] ..............................................................23

CHNG III: KT QU V THO LUN..........................................................26 III.1 XC NH CC CH TIU HA L CA TINH DU BC H.............26

III.1.1 nh gi cm quan..................................................................................26 III.1.2 Xc nh ch s acid (IA).........................................................................26 III.1.3 Xc nh ch s savon ha (IS) ................................................................27 Bng 7: Kt qu chun xc nh ch s savon v ch s ester.........................27

III.2 KHO ST CC YU T NH HNG N HIU SUT V THNH PHN TINH DU BC H TRONG QU TRNH LY TRCH..........................27

III.2.1 Thi gian ly trch....................................................................................27 III.2.2 Kho st nhit ly trch........................................................................29 III.2.3 Kho st lng dung mi ly trch ...........................................................30 III.1.4 Kho st s nh hng ca thi gian, nhit v th tch dung mi n thnh phn ha hc tinh du Bc h ...................................................................31

III.3 XC NH THNH PHN HA HC CA TINH DU BNG PHNG PHP gc ms .......................................................................................................34

III.3.1 Thnh phn ha hc ...............................................................................34 III.3.2 So snh kt qu nghin cu ....................................................................37

CHNG IV: KT LUN V KIN NGH..........................................................39 IV.1 KT LUN ..................................................................................................39 IV.2 KIN NGH..................................................................................................39 PH LC PH GC MS .....................................................................................41

TI LIU THAM KHO .........................................................................................45

PH LC HNH

ix

PH LC HNH ------------

Hnh 1: V tr trong phn loi thc vt ca Bc h Mentha arvensis L..........................2

Hnh 2: L v cy Mentha arvensis L. .........................................................................3

Hnh 3: Thn v hoa Mentha arvensis L. .....................................................................3

Hnh 4: Tinh du Bc h ..............................................................................................6

Hnh 5: H thng Soxhlet ..........................................................................................12

Hnh 6: Mu Bc h kh ............................................................................................17

Hnh 7: Mu Bc h ct nh .................................................................................17

Hnh 8: S cc bc thc hin ly trch tinh du Bc h bng phng php chng

ct li cun hi nc .................................................................................................19

Hnh 9: Chng trnh nhit ca GC - MS chy phn tch thnh phn ha hc tinh

du Bc h .................................................................................................................24

Hnh 10: iu kin chy ca MS ...............................................................................24

Hnh 11: iu kin Injecter........................................................................................25

Hnh 12: Tinh du bc h..........................................................................................26

Hnh 13: th biu din thi gian ly trch theo hm lng tinh du Bc h thu

c ..........................................................................................................................28

Hnh 14: th biu din nhit ly trch theo hm lng tinh du Bc h thu c 30

Hnh 15: th biu din th tch dung mi ly trch theo hm lng tinh du Bc h

thu c ....................................................................................................................31

Hnh 16: Cng thc ca mt s thnh phn chnh trong tinh du Bc h....................34

Hnh 17: Ph phn tch thnh phn tinh du Bc h bng GC - MS.......................34

Hnh 18: Khi ph ca menthol .................................................................................35

Hnh 19: Khi ph ca menthol cung cp t th vin ph NIST ...............................35

Hnh 20: S phn mnh ca phn t menthol ........................................................36

PH LC BNG

x

PH LC BNG ------------

Bng 1: Cc ch s l ha ca tinh du Bc h ..........................................................6

Bng 2: Thnh phn ha hc chnh ca tinh du Mentha piperita L. xc nh bng

phng php GC MS. ...............................................................................................8

Bng 3: Kt qu hot tnh khng khun (MIC, mg/ml).................................................8

Bng 4: Thnh phn ha hc ca Bc h Mentha arvensis L.[15] ................................10

Bng 5: Thnh phn ha hc ca Bc h Mentha piperita L.[15].................................11

Bng 6: Kt qu chun xc nh ch s acid ..........................................................26

Bng 7: Kt qu chun xc nh ch s savon v ch s ester.................................27

Bng 8: Kt qu kho st thi gian ly trch tinh du bc h bng phng php chng

ct li cun hi nc .................................................................................................28

Bng 9: Kt qu kho st nhit ly trch tinh du bc h bng phng php chng ct

li cun hi nc.......................................................................................................29

Bng 10: Kt qu kho st th tch dung mi ly trch tinh du bc h bng phng

php chng ct li cun hi nc ..............................................................................30

Bng 11: Bng so snh thnh phn ha hc ca 4 mu TIME TU, VOL TU, TEM

TU v MAU KS tinh du bc h bng phng php chng ct li cun hi nc 32

Bng 12: Thnh phn chnh trong tinh du Bc h.....................................................36

Bng 13: So snh kt qu nghin cu thnh phn Bc h Mentha arvensis L. Vit Nam

v Mentha arvensis L. Himalaya................................................................................37

Bng 14: Kt qu iu kin ti u cho qu trnh ly trch ...........................................39

Bng 15: Cc ch s ha hc ca tinh du Bc h ......................................................39

PH LC BNG

xi

DANH SCH CC T VIT TT ------------

GC MS Gas chromatographymass spectrometry

IA Acid index

IS Savon index

IE Ester index

FID Flame ionization detector

MIC Minimum inhibitory concentration

APG III Angiosperm Phylogeny Group III system

LI M U

xii

LI M U ------------

Trc chin tranh th gii th nht, Nht Bn c xem l nc dn u trong

sn sut cc sn phm thng mi t cy Bc h nh: dc phm, kem nh rng,

nc xc ming, ko thuc l, m phm, trn th trng th gii. Thi gian ny

c xem l thi k thnh vng ca cy Bc h Mentha arvensis Nht Bn v n

cn c gi l Japani mint. Sau chin tranh th gii th hai, Nht Bn chuyn sang

trng cy Bc h phc thng mi v xut khu sang Brazin v Bc M. Cng thi

gian ny, n , Trung Quc, M, v nhiu nc khc cng c nhng thnh tu ni

bt trong sn sut cc sn phm t Bc h. Nm 1960, n l nc sn sut tinh du

Bc h ln nht th gii. Trung Quc lc ny cng c xem l ngun cung cp

menthol chnh cho th trng thi gii.

Bn cnh nhng gi tr kinh t mang li, th Bc h cn c mt vai tr quan trng

trong ng Y l cha tr cc bnh nh cm st, au nhc bn chn, vt cn trng

cn[1], Nhng a phn Bc h dng cha tr l s dng hn hp ton phn hoc

theo kinh nghim dn gian. V th vic nghin cu thnh phn ha hc cy Bc h

tm nhng hp cht c cu trc mi hoc nhng cht c tc dng chnh s dng

di dng tinh khit cng nh trong iu tr bnh l vic lm cn thit.

Chnh v cy Bc h c nhiu cng dng nh vy nn rt sm c nhiu cng

trnh nghin cu v cy ny nhng a s l ngoi nc. Nhm gp phn lm r hn

thnh phn ha hc ca cy Bc h, nn em chn ti Ly trch v kho st thnh

phn ha hc ca tinh du Bc h (Mentha arvensis L.) gp phn gip cho vic

bo ch v s dng c thun li hn. ng thi nng cao gi tr s dng ca cy

Bc h. Trong gii hn nghin cu ca ti em tin hnh nghin cu mt s vn

sau:

Nghin cu iu kin ly trch ti u ca tinh du Bc h bng phng php

chng ct li cun hi nc.

Xc nh cc ch tiu ha l ca tinh du Bc h.

Xc nh thnh phn ha hc chnh trong tinh du Bc h bng phng php

GC MS.

CHNG I: TNG QUAN

1

CHNG I

TNG QUAN

I.1 GII THIU H LAMIACEAE

H Hoa mi (danh php khoa hc: Lamiaceae hay Labiatae), cn c gi bng

nhiu tn khc nh h Hng, h Bc h [1,2] l mt h thc vt c hoa. N tng c

coi l c h hng gn vi h Verbenaceae nhng mt s nghin cu pht sinh loi gn

y ch ra rng mt lot cc chi c phn loi trong h Verbenaceae thc cht l

thuc v h Lamiaceae, trong khi cc chi ct li ca h Verbenaceae th khng c

quan h h hng gn vi Lamiaceae m l c quan h h hng gn hn vi cc thnh

vin khc ca b Lamiales. H Lamiaceae c khong 200 chi v khong 3500 loi

(theo h thng phn loi APG III)[1].

Trong hu ht cc phn ca thc vt trong h ny u c hng thm v bao gm

nhiu loi cy thn tho c s dng rng ri trong m thc, nh hng qu, Bc h,

hng tho, x thm, hng Bc h, kinh gii, ngu ch, bch l hng, oi hng,

ta t, hng nhu. Mt s loi l cy bi hay cy g, him gp hn l cc dng dy leo.

Nhiu loi c gieo trng rng ri, khng ch v hng thm ca chng m cn v d

gieo trng, d nhn ging nht bng cc gim cnh. Bn cnh nhng loi ly l n,

lm gia v cn mt s loi c trng lm cnh nh hng chanh. Mt s loi khc

c trng v mc ch ly ht (ch khng phi l) lm thc phm, nh ht cy

Chia[1,2].

Tn gi nguyn gc ca h ny l Labiatae, do hoa ca chng thng thng c

cc cnh hoa hp thnh mi trn v mi di. Tn gi ny hin nay vn l hp l,

nhng phn ln cc nh thc vt hc hin ti thch s dng tn gi "Lamiaceae" hn

khi ni v h ny.

CHNG I: TNG QUAN

2

I.1.1 Phn loi hc[1,2]

Hnh 1: V tr trong phn loi thc vt ca Bc h Mentha arvensis L.

Bc h l cch gi ca ngi Vit ta. mt s a phng khc cn c cch gi

khc l Bc h nam. y l loi ph bin nht nc ta. C tn khoa hc l Mentha

arvensis L. Ty theo tng khu vc kh hu mi nc cng nh mi chu lc m Bc

h c c im v thnh phn tinh du khc nhau. Do , chi Bc h (Mentha) c

phn loi vi hn hng trm loi, vi cc loi ph bin nh:

+ Mentha piperita L. (Bc h u) cho tinh du vi tn thng phm l

peppermint oil (Oleum Menthae piperita), thnh phn chnh ca tinh du l

menthol (50-68%).

+ Mentha arvensis L. (Bc h ) cho tinh du vi tn thng phm l Cornmint

oil (Oleum Menthae arvensis), thnh phn chnh ca tinh du l menthol (>70%).

Gii

Ngnh

Lp

H

Loi

Plantae

Phn loi thc vt

Magnoliophyta

Magnoliopsida

Lamiales

Lamiaceae Lindl.

Mentha L.

B

Chi

M. arvensis

CHNG I: TNG QUAN

3

+ Mentha spicata L. (Bc h bng) cho tinh du vi tn thng phm l Native

Spearmint, thnh phn chnh ca tinh du l carvon (>50%).

+ Mentha cardiaca Grard ex Baker (Bc h bng) cho tinh du vi tn thng

phm l Scotch Spearmint, thnh phn chnh ca tinh du l carvon (>50%).

+ Mentha citrate Ehrh., (Bc h chanh) cho tinh du vi tn thng phm l

Bergamot mint oil, thnh phn chnh ca tinh du l linalyl acetat (33-74%) v linalool

(25-52%).

+ Mentha pulegium L. (Bc h bng) cho tinh du vi tn thng phm l

Pennyroyal oil, thnh phn chnh ca tinh du l pulegon (80%).

I.1.2 M t[1,2]

Hnh 2: L v cy Mentha arvensis L. Hnh 3: Thn v hoa Mentha arvensis L.

(Ngun: http://blog.daum.net) Bc h (Mentha arvensis L.) l mt loi cy tho sng lu nm. Thn mm, hnh

vung. Loi thn ngm mang r mc b lan, loi thn ng mang l, cao t 0,30-

0,40m, c th cao ti 1m, mu xanh lc hoc tm ta, i khi phn nhnh. L mc

i, cun ngn, phin l hnh trng hoc hnh bu dc, mp c kha rng u, mt

trn v mt di u c lng che ch v lng bi tit.

Hoa mc vng k l, cnh hoa hnh mi mu tm hay hng nht c khi mu

trng, trng c ng ngn, phin trng chia lm bn phn bng nhau, c mt vng lng

pha trong. Bn nh bng nhau ch nh nhn. t khi thy c qu v ht.

Ton thn c lng che ch v lng tit tinh du v c mi thm. Ma hoa qu

vo thng 7-10.

I.1.3 Phn b v thu hi[1,2]

Cy Bc h nc ta thng mc hoang, v mt s c trng ly l v cy

lm thuc. Gn y, Bc h c trng nhiu ly tinh du nh lng Ngha
http://blog.daum.net/

CHNG I: TNG QUAN

4

Trai Hng Yn; i Yn, Gia Lm H Ni; i hc Dc H Ni; Sn La, Sapa,

Ph Quc, Tam o, Tp H Ch Minh

C hai ma trng Bc h cho nng sut cao l vo ma xun (thng 2-3) v ma

thu (thng 8-9). Tt nht l trng vo ma xun cho nng sut cao nht. Sau 3-4 thng

gim trng c th thu hoch t u. Sau cn vun xi, bn phn v ti nc

thng th nhng t thu hoch tip theo c cch 2 thng mt ln[1].

I.1.4 Cng dng ca Bc h

T xa xa, cy Bc h c bit n l mt v thuc rt hu hiu dng

cha tr cc bnh tiu ha v cha tr cc bnh ng rut. Bc h cn cha tt cc

bnh cm st, gip long m, thng mi, mt hng, gim cng thng v mt ng.

Ngoi ra, tinh du Bc h cn c s dng lm p da, gim cc vt tho v cc

vt thm mn, ri c tc dng tr gu na[1]. Do menthol trong tinh du Bc h gy ra

cm gic mt lnh, nn gip lu thng mu lm tinh thn thng thong v sng khoi,

tng cng h min dch. Nhng nu dng vi mt lng nhiu n c th gy t, do

tinh du Bc h cn c s dng nh mt cht gy m, lm gim au[1].

Trong cuc sng hin i ngy nay, Bc h v tinh du Bc h cn c ng

dng vo rt nhiu lnh vc khc nhau nh:

Trong thc phm, l bac c dng trang tr trn thc n hay c dng nh

mt loi rau n km thay th rau thm, rau hng nhi. L Bc h cn c th c xy

nhuyn ly nc lm nc st Bc h. Ngoi ra, Bc h c th dc nu ly

phn nc mu xanh dng lm xi r, nc gii kht v Bc h, lm bnh kem, ko

Trong m phm, tinh du Bc h rt c gi tr trong vic iu tr da b mn, nh

tnh khng khun cao. ng thi li lm mt da v lm sng cc vng da b sm mu.

i vi da nhn, th Bc h cn c tc dng lm da bt bng nhn, sn chc m li

khng b kh da. Mt nghin cu gn y nghin cu ca trng i hc Wheeling

Jesui cn cho thy rng nhng ngi ht tinh du Bc h trong vng hai ting s np

lng calorie t hn nhng ngi khng ngi hng Bc h 23%. Do , tinh du Bc

h cn c tc dng lm gim cn. Ngoi ra, cht menthol trong tinh du Bc h cn l

thnh phn chnh trong cc sn phm kem nh rng, du gi v mt s loi nc hoa.

Trong dc phm, tinh du Bc h c s dng ch yu kt tinh thu menthol.

T iu ch nhng dc phm m da vo cng dng ca menthol mang li nh:

CHNG I: TNG QUAN

5

ming dn gim au, gim st, thuc thoa tr trt khp, bong gn, thuc nh mt, thuc

tr ho, au hng v du gi

Trong thuc dit cn trng, tinh du Bc h ng vai tr nh cht c ch i vi

mt s loi cn trng nh: mui, kin, gin, ong bp cy. Do , tinh du Bc h c

ng dng xua ui chng.

I.2 TINH DU

I.2.1 Khi qut v tinh du

Tinh du l mt hn hp nhiu cht d bay hi, c mi c trng ty thuc vo

ngun gc nguyn liu cung cp tinh du. Phn ln tinh du c ngun gc t thc vt

v s t t ng vt. Trong thin nhin tinh du trng thi t do, ch c mt s t

trng thi tim tng, ngha l tinh du khng c sn trong nguyn liu m ch xut hin

trong nhng iu kin gia cng nht nh trc khi tin hnh ly trch hay di tc

dng c hc. Cn trng thi t do, tinh du c sn trong nguyn liu c th thu hi ly

trch trong iu kin bnh thng[3].

I.2.2 Qu trnh tch ly[3,4,5]

Trong thc vt tinh du c to ra v tch ly trong cc m. Hnh dng cc m

ny thay i ty theo v tr ca chng trong cy. Nhng m ny c th hin din tt

c cc b phn ca cy nh r, thn, l, hoa v c trivi nhng tn gi khc nhau

nh:

T bo tit: tinh du c tit ra ri chng c gi trong t bo (m tit) v d

trong cnh hoa hng, trong c gng

Lng tit: cng l t bo tit nhng nm nh ra ngoi thc vt, thng bt gp

cc loi hoa mi, cc, c

Ti tit: t bo tit ra tinh du nhng khng cha li bn trong m dn chung

cha vo mt xoan trng, to ra bi c ch ly bo hay tiu bo. Ti tit thng nm

bn di lp biu b. Thng c cc ging Cirtrus, eucalyptus

ng tit: cch to ra tinh du cng ging nh ti tit nhng nm su trong phn

g v chy di theo s g, thng bt gp trong cc ging Dipterocarpi, Artemisia

CHNG I: TNG QUAN

6

I.2.3 Tinh du Bc h[3]

Hnh 4: Tinh du Bc h

(Ngun: http://tinhdauthomtv.com)

L mt cht lng c mu vng nht, mi Bc h, thanh mt v c v t. Thu c

bng cch chng ct li cun hi nc l Bc h.

Thnh phn, tnh cht ca tinh du Bc h ty thuc vo ging, t ai, kh

hu... Trn th gii hin nay c cc ging Bc h chnh nh sau: Nga, Anh, M,

Php, Trung Quc v Nht Bn. Loi tinh du Bc h tt nht hin nay l t Bc h

Nht Bn (Mentha arvensis L.).

Cc ch s l ha ca tinh du Bc h Mentha arvensis L. nh sau:

Bng 1: Cc ch s l ha ca tinh du Bc h

Thng s 20

d 20Dn []D IA IS IE

Gi tr 0,895 1,4580 -24o 2 11 9

Trong :

20d : t trng ca tinh du Bc h.

20D

n : chit sut ca tinh du Bc h.

[]D : quay cc ca tinh du Bc h.

IA : ch s acid ca tinh du Bc h.

IS : ch s savon (ch s x phng ha) ca tinh du Bc h.
http://tinhdauthomtv.com/

CHNG I: TNG QUAN

7

IE : ch s ester ca tinh du Bc h.

Thnh phn ch yu l menthol (69 92%), menthone (3 6,9%). Ngoi ra cn

cha menthyl acetate, L-limonene, piperitone, pulegone...

Cht lng ca tinh du ph thuc vo phng php x l sau khi chng ct v

chia lm 3 loi:

- Tinh du th l tinh du khng qua x l.

- Tinh du tinh luyn mt ln.

- Tinh du tinh luyn hai ln.

Tinh du tinh luyn hai ln c cht lng tt nht v cn gi l menthol th.

I.3 TNH HNH NGHIN CU TINH DU BC H

I.3.1 Kho st hot tnh khng vi sinh vt ca tinh du Bc h Mentha

piperita L.[13]

Cng trnh c ng trn t bo Journal of Agricultural and Food Chemistry do

nhm nghin cu thuc Trng i hc Anadulo Th Nh K.

Cng trnh nghin cu c thc hin trn loi Bc h Mentha piperita L. Nhm

nghin cu tin hnh xc nh thnh phn ca tinh du Bc h trn cng mt loi c

trng 4 a im khc nhau: (A) Mentha piperita oil (Evcin company, Turkey), (B)

Mentha piperita oil (Jet Farms, Yakima), (C) Mentha piperita oil (Mari Linn Farms,

Oregon), (D) Mentha piperita oil (Erdomus Perfume Industry, imported from India)

bng sc k kh ghp khi ph (GC MS). Sau tin hnh kho st hot tnh sinh hc

ca 4 loi tinh du ny v so snh kt hp vi hot tnh sinh hc ca menthol v

menthone bng k thut in vitro. Kt qu thu c nh sau:

Thnh phn ha hc chnh trong tinh du Bc h Mentha piperita L.

CHNG I: TNG QUAN

8

Bng 2: Thnh phn ha hc chnh ca tinh du Mentha piperita L. xc nh bng phng php GC MS.

Thnh phn A B C D

Limonene 1,9 1,3 1,0 2,1

1,8-Cineole 5,3 4,0 3,4 4,0

Menthone 27,9 21,2 27,2 18,4

Menthofuran 5,5 3,8 1,3 3,2

Isomenthone 3,5 2,9 3,8 2,9

Linalool 2,5 4,4 4,1 4,8

-Caryophyllene 4,2 1,8 1,6 1,5

Terpinen-4-ol 1,2 3,4 3,8 2,1

Menthol 27,5 42,3 39,9 34,6

Pulegone 6,4 1.0 1,9 14,4

-Terpineol 2,4 1,6 1,7 0,7

Hot tnh sinh hc ca tinh du Bc h Mentha piperita L.

Bng 3: Kt qu hot tnh khng khun (MIC, mg/ml)

Vi sinh vt A B C D M1 M2

Escherichia coli 2,5 1,25 1,25 1,25 1,25 5,0

Staphylococcus aureus 0,625 0,625 1,25 2,5 0,625 2,5

Pseudomonas aeruginosa 2,5 2,5 5,0 2,5 2,5 5,0

Enterobacter aerogenes 1,25 1,25 1,25 2,5 1,25 5,0

Proteus vulgaris 2,5 5,0 1,25 1,25 1,25 2,5

Salmonella typhimurium 1,25 1,25 1,25 2,5 0,625 5,0

Klebsiella pneumoniae 2,5 2,5 2,5 2,5 2,5 5,0

Yersinia enterocolitica 2,5 2,5 2,5 2,5 2,5 2,5

Listeria monocytogenes 0,156 0,625 2,5 0,312 0,625 1,25

Bacillus cereus 1,25 1,25 1,25 1,25 1,25 1,25

Staphylococcus

epidermidis 0,625 2,5 0,625 1,25 0,625 0,625

CHNG I: TNG QUAN

9

Xanthomonas

campestrispv.phaseoli 0,625 0,312 0,625 0,156 0,625 2,5

Pseudomonas

syringaepv.phaseolicola 2,5 1,25 1,25 0,625 1,25 2,5

Pseudomonas

syringaepv.tomato 0,07 0,07 0,625 0,312 0,07 1,25

Pseudomonas

syringaepv.syringae 0,312 0,156 0,312 0,312 0,156 2,5

Xanthomonas

campestrispv.campestris 0,156 0,156 0,312 0,156 0,156 1,25

Candida albicans 0,625 0,625 0,312 0,625 0,625 2,5

Trong :

M1 : (-)-menthol

M2 : (-)-menthone

I.3.2 Thnh phn tinh du ca Bc h Mentha arvensis L. v Bc h

Mentha piperita L. nhng giai on sinh trng khc nhau trong vng

Kumaon thuc pha ty Himalaya.

Th nghim c tin hnh kho st trn hai loi ging: Bc h Mentha arvensis L.

cc khu vc Kosi, Saksham, Alka v Himalaya; loi th hai l Bc h Mentha

piperita L. trng khu vc Kukrail, CIM-Madhurus v CIM-Indus. Ging Bc h

Mentha arvensis L. c trng vo thng ba v ging Bc h Mentha piperita L.

c trng vo thng ging. Cc ging c thu hi sau khi c 90 ngy tui (tnh

t lc bt u gieo trng). Tin hnh ly trch tinh du trn mu ti sau khi c

thu hi bng phng php chng ct li cun hi nc.

Sau tinh du ca hai ging Bc h thu c s c phn tch thnh phn

ha hc bng phng php sc k kh s dng u d FID. Ct s dng l ct mao

qun silica (30 m x 0.25 mm x 0.25 m), pha tnh l BP-20, kh mang s dng l

kh hidro. Kt qu thnh phn tinh du thu c nh sau:

CHNG I: TNG QUAN

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Bng 4: Thnh phn ha hc ca Bc h Mentha arvensis[15]

Thnh phn Kosi Saksham Himalaya Kalka

-Pinene 0,47 0,61 0,55 1,25

-Pinene 0,56 0,79 0,24 0,86

Sabinene T T 0,14 T

-Myrcene 0,40 0,40 0,31 0,49

-terpinene - - - -

Limonene 1,57 0,98 0,67 1,85

1,8- cineole T T 0,11 -

-terpinene - - - -

(E)- -ocimene - - - -

(Z)-3-hexanol - - - -

3-octanol 1,61 4,24 1,73 T

Menthone 7,46 6,53 5,08 11,85

(E)-sabinene hydrate - T T -

Iso-menthone 3,56 3,48 3,16 6,09

Linalool T - - -

Menthyl acetate 1,12 1,44 1,63 3,67

Isopulegol 1.01 0,94 0,73 1,16

-Caryophyllene - - - -

Neo-menthol 1,93 1,74 1,61 1,45

Terpinen-4-ol 0,65 0,33 0,31 0,40

Pulegone T 0,34 T T

Menthol 77,54 75,85 79,64 65,61

Isomenthhol 0,15 0,11 T T

-terpineol - 0,17 0,17 0,18

Germacrene D 0,16 T 0,14 0,31

Piperitone 0,24 1,76 1,08 2,05

Cavone - - - -

CHNG I: TNG QUAN

11

Bng 5: Thnh phn ha hc ca Bc h Mentha peppermint[15]

Thnh phn Kukrail CIM-Madhurus CIM-Indus

-Pinene 0,91 1,00 0,88

-Pinene 0,55 1,48 T

Sabinene - 0,22 1,64

-Myrcene 0,24 0,26 0,42

-terpinene 0,21 0,18 15,46

Limonene 2,37 1,54 0,51

1,8- cineole 5,58 6,57 5,96

-terpinene 0,55 0,35 13,94

(E)- -ocimene 0,30 1,29 3,65

p-cimene 0,37 0,18 1,38

3-octanol T T 0,43

Menthone 29,83 26,88 0,77

Menthofurane 0,81 0,32 7,45

Iso-menthone 3,36 2,65 1,37

Linalool 0,19 0,26 0,34

Menthyl acetate 5,18 7,23 1,23

Isopulegol 0,13 0,13 0,27

-Caryophyllene 0,96 1,10 4,13

Neo-menthol 3,47 3,39 0,38

Terpinen-4-ol 0,51 1,21 2,07

Pulegone 0,68 0,96 T

Menthol 37,44 36,13 22,56

(E)- -farnesene 0,18 0,21 0,40

Isomenthhol 0,19 0,25 T

-terpineol 0,16 0,16 0,44

CHNG I: TNG QUAN

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Germacrene D 0,78 0,69 2,49

Piperitone 1,27 0,83 1,19

Cavone - - 0,52

T : nhng cht c hm lng < 0,1%.

I.3.3 Tch v tng hp menthol

Trong cng nghip nguyn liu chnh tch menthol l tinh du Bc h.

tch menthol ra khi tinh du Bc h ngi ta lm lnh tinh du, sau li tm s

c menthol th. Chng ct phn on tinh du Bc h v ly on c nhit si

98-100oC/10,5 mm. ng thi vi menthol cn c neomenthol. tch ring

neomenthol ra khi menthol, cho hn hp tc dng vi acid xucxenic hoc acid

phtalic, neomenthol, s to thnh ester vi hai acid trn v tch ring ra khi menthol.

Menthol cn c th tng hp c bng cch kh menthone, kh pulegone hay hydro

ha timol. Nhng phng php c hiu qu kinh t nht l hydro ha p cymene theo

s sau.

I.4 MT S PHNG PHP LY TRCH TINH DU BC H

Da trn cch thc hnh, cc phng php sn xut tinh du c chia ra lm 4

loi: c hc, tm trch, hp th v chng ct hi nc.

Nhng d c tin hnh theo bt c phng php no, quy trnh sn xut u c

nhng c im chung sau y[4,5].

Tinh du thu c phi c mi thm t nhin nh nguyn liu.

CHNG I: TNG QUAN

13

Quy trnh khai thc phi ph hp vi nguyn liu.

Tinh du phi c ly trit khi nguyn liu, vi chi ph thp nht.

Nguyn tc ly trch ca tt c cc phng php ni trn u da vo nhng c

im ca tinh du nh:

D bay hi.

Li cun theo hi nc nhit ph hp.

Ha tan d dng trong dung mi hu c.

D b hp th ngay th kh.

I.4.1 Phng php c hc[5,6]

I.4.1.1 Vt p

Ngm nguyn liu vo nc thng hoc nc mui 5%. Vt nguyn liu ra, vt

cho tinh du tun ra, ri thm phn tinh du ny bng ming bng hoc bt b, vt ro

ming bng ly tinh du ra ri tip tc thao tc nh trn. Sau cng tch nc lm

khan v lc sch.

I.4.1.2 No xt

Dng mt phu bng ng, mt trong c cc gai nh, dng nguyn liu xt ln

mt phu lm cho cc ti du v ra. Tch nc, lm khan, thu ly tinh du.

I.4.1.3 p

Va p nguyn liu va phun nc mui 10% tinh du d tri ra v ng thi

khi ngng p, tinh du khng b p tr li xc. em dung dch li tm hoc lng gn

thu phn tinh du.

Cc phng php c hc ny, c u im l tinh du c mi thm t nhin do

khng dng n sc nng v dung mi, nhc im l tn nhiu nhn cng v i hi

cy trng tp trung, hiu sut khng cao, tinh du ly ra khng trit , do phi

dng phng php khc ly lng tinh du cn li.

I.4.2 Phng php dng dung mi ha tan[5,6]

Phng php ny phi dng mt s dung mi thch hp nh ethanol, ether du

ha d ha tan tinh du c trong nguyn liu.

CHNG I: TNG QUAN

14

Cho nguyn liu vo bnh c nt kn, thm dung dch vo ngm trong 8 ngy.

Trong thi gian ngm thnh thong lc bnh cho u, sau lc v p. Ta s c dung

dch mi ca nguyn liu thin nhin. Vi loi tinh du ny ta c th pha ch nc

hoa, ko, bnh

u im ca phng php ny l thu c tinh du c mi thm t nhin, c

hiu sut cao.

Nhc im phng php ny l khng mang tnh kinh t, dung dch ethanol thu

c thng c ln mu xanh ca dip lc t nn khi pha ko hoc nc ung mu sc

cng b nh hng.

I.4.3 Phng php chng ct li cun hi nc[4,5,6]

I.4.3.1 Phng php c in

Nguyn liu c cho vo ni ct. Sau , cho chy qua nguyn liu mt lung

hi nc, hoc cho nc trc tip vo nguyn liu ri un si. Hi nc v hi tinh

du c ko sang bnh lm lnh, ng li thnh cht lng, ch mt phn rt nh tinh

du tan trong nc. Tip theo dng mt bnh ring gi l florentin gn ly tinh du.

Phn tinh du thu c c ln nc s c chit li vi diethyl ether, sau lm khan

nc bng N2SO4 khan. C quay ui dung mi ta thu c tinh du cn ly. Phng

php ny tng i kinh t hn, t tn cng, hiu sut thu hi tinh du cao. Nhng tinh

du c mi khng t nhin v khng th p dng cho nhng loi tinh du d b bin

tnh bi nhit .

I.4.3.2 Phng php ly trch kt hp vi sng[4,6]

Mt s phn t, trong c nc, phn chia in tch trong phn t mt cch bt

i xng. Do , khi t trong in trng mt chiu, cc phn t ny chuyn ng

nh nhng lng cc nh hng theo chiu ca in trng. Nu l in trng xoay

chiu, th s nh hng ca cc lng cc s thay i theo chiu ca in trng .

Phn t nc c phn cc ln, nn nc l mt cht rt l tng un nng

bng vi sng. Vi sng c p dng trong ha hc ch yu l khai thc hin tng lm

nng ln ca vt cht. Nhng phn t ny c lng cc nh hng theo chiu ca t

trng, nn in xoay chiu c tn s cao s gy mt xo ng rt ln ln cc phn t

trn khin chng b ma st mnh, y chnh l s nng ln ca vt cht.

CHNG I: TNG QUAN

15

Tm li: vi sng cung cp mt phng php duy nht v s un nng khng

dng s truyn nhit bnh thng, tc l sc nng i t b mt ca vt cht vo bn

trong, lm nng vt cht t bn trong n. S un nng ny rt hiu qu, thi gian

nhanh v c bit bo v c cc hp cht d b phn hy nhit.

ng dng ca vi sng vo ly trch tinh du thc vt

Di tc dng ca vi sng, nc trong cc t bo thc vt b nng ln, p sut

bn trong tng t ngt lm cc m cha tinh du b v ra.

Tinh du thot ra bn ngoi, li cun theo hi nc sang

h thng ngng t (phng php chng ct hi nc)

hoc ha tan vo dung mi hu c ang bao ph bn

ngoi nguyn liu.

I.4.4 Phng php chit Soxhlet[5]

H thng my Soxhlet gm ba b phn chnh c

ni vi nhau l: bnh cu cha dung mi c nung trn

bp in c th iu chnh nhit ; phn gia l b

phn cha mu bt cy, b phn ny gm ba ng thy tinh thng vi nhau; phn trn

cng l h thng ngng tu hi.

Mu c cho vo ti v vo ng D. Sau cho dung mi vo, dung mi s

thm t bt cy ri chy xung bnh cu theo ng thng nhau E. Lu dung mi

vo bnh cu khng c qu 2/3 th tch bnh. Lc ny m nc chy hon lu v

tin hnh un. Hi dung mi bc ln s theo ng B, gp lnh hi s bi ngng tu

xung ng D. Dung mi thm vo bt cy v chit nhng cht hu c no c th ha

tan vo dung mi. Khi dung mi ngng tu n mc cao nht trong ng E th ton b

dung mi b rt ht xung bnh cu. Cc cht c ht xung nm li ti v ch c

dung mi bay hi ln tip tc qu trnh chit. Sau khi hon tt ui dung mi ta thu

c cao chit.

Hnh 5: H thng Soxhlet

CHNG II: THC NGHIM

16

CHNG II: THC NGHIM

II.1 A IM, THI GIAN, THIT B, NGUYN LIU V

HA CHT

II.1.1 a im v thi gian

Vic tin hnh nghin cu c thc hin ti phng th nghim Ha sinh, B

mn Ha hc, Khoa Khoa hc T nhin, Trng i hc Cn Th

Thi gian thc hin: t 01/12/2012 n 30/05/2013

II.1.2 Thit b, nguyn liu, dng c v ha cht

II.1.2.1 Thit b v dng c

B chng ct tinh du nh Clevenger

Cn in t SARTORRIUS 210 g 0,0001 g

H thng GC MS THERMO SCIENTIFIC

My c quay EYELA CCA - 1110

Nhit k 200C

Bnh tam gic 250 ml

Bercher 100ml, 250 ml

Microburrette 25 ml

a thy tinh

Pipet 5 ml, 10 ml, 25 ml

Bnh cu 250 ml

ng ong 10 ml, 25 ml

Bnh lng 250 ml

Bp in Gali

Bp un hon lu Heating Green

CHNG II: THC NGHIM

17

II.1.2.2 Nguyn liu v ha cht

Nguyn liu

Hnh 6: Mu Bc h kh Hnh 7: Mu Bc h ct nh

Mu Bc h dng trong th nghim l mu l Bc h kh do c s ging cy

trng Bnh Chu cung cp (24 - in Bin Ph, qun Bnh Tn, TP. HCM).

Qua s nh danh B mn Sinh hc Khoa Khoa hc T nhin, Trng i hc

Cn Th v tham chiu trong quyn Nhng cy thuc v ng vt lm thuc Vit

Nam loi Bc h c s dng kho st thc nghim c tn gi khoa hc l Mentha

arvensis L. var. piperascens (Malinv.)

Ha cht

Cn 96o (25 ml)

Nc ct

Acetone (50 ml)

Phenolphthalein (5 ml)

Dung dch KOH 0,1N trong alcol (100 ml)

Dung dch HCl 0,1N (200 ml)

bt (5 g)

Diethyl eter

II.2 PHNG PHP NGHIN CU

t c yu cu nghin cu ca ti, em tin hnh thc hin cc bc sau:

- Thu mua v x l nguyn liu

- Ly trch tinh du bng phng php chit li cun hi nc vi cc iu kin

kho st l thi gian ly trch, nhit ly trch v lng dung mi ly trch.

- Xc nh cc ch tiu ha l ca tinh du.

CHNG II: THC NGHIM

18

- Phn tch thnh phn ha hc chnh trong tinh du bng my sc k ghp khi

ph (GC MS).

II.2.1 X l nguyn liu

Mu Bc h kh sau khi mua v c loi b tp ln nh c, rm hoc cc l cy

khc. Tip theo tch ly l Bc h, sau ct nh mu khong 1-2 cm v c bo

qun trong ti nilon nhit phng.

II.2.2 Ly trch tinh du bng phng php li cun hi nc[7]

Cn 20 g mu Bc h kh ct nh cho vo bnh cu 1000 ml, rt nc ct vo

bnh sao cho th tch nc trong bnh khng vt qu 2/3 th tch ca bnh cu. Nu

cho nc qu nhiu th trong qu trnh un xc mu trong bnh cu s tro ln b phn

hng ly tinh du. Lp bnh cu vo h thng gm ng hng tinh du v ng sinh hn.

Bnh cu c un cch du silicon nhit ta u, n nh hn v khng gy v

bnh cu khi un qu lu. iu chnh nhit ca bp in sao cho trnh du si mnh

c th trn ra ngoi. Sau qu trnh ly trch, hn hp tinh du thu c gm tinh du

Bc h v nc. Thc hin chit hn hp vi dung mi diethyl eter loi b nc thu

ly phn dung mi. Lm khan bng Na2SO4, sau tin hnh c quay thu hi dung

mi, c tinh du Bc h. Cn sn phm thu c t tnh hiu sut ca qu trnh

ly trch. ng thi cng phn tch thnh phn mu tinh du thu c bng GC MS.

CHNG II: THC NGHIM

19

Hnh 8: S cc bc thc hin ly trch tinh du Bc h bng phng php chng ct li cun hi nc

II.2.3 Xc nh mt s ch tiu ha l ca tinh du Bc h

Mu tinh du thu c sau khi ly trch bng phng php chng ct li cun hi

nc s c xc nh cc ch tiu sau:

Nguyn liu

Ly trch bng li cun hi nc

Hn hp tinh du

Nc Dung mi

C quay thu hi diethyl ether

Dung mi

Nc

Tinh du bc h th

+ Lm khan bng Na2SO4 + Lc

+ Chit li hn hp vi diethyl ether

+ Lm khan bng Na2SO4 + Lc + C quay thu hi dung mi

+ Chit hn hp vi diethyl ether loi b tp cht

+ Loi b c, l cy tp + Ct nh

CHNG II: THC NGHIM

20

II.2.3.1 nh gi cm quan[6,7]

Phn tch s b cht lng ca tinh du bng cm quan. Ngha l nghin cu

nhng du hiu bn ngoi nh mi, v, mu sc, trong sut...

Qua nghin cu nhng du hiu bn ngoi ny c th phn on v nh gi s

b v cht lng ca tinh du v mc ch s dng tinh du.

II.2.3.1.1 Mu sc v trong sut[6,7]

Xc nh mu sc v trong sut ca tinh du bng cch cho tinh du vo mt

ng thy tinh trong sut khng c mu c dung tch 20 ml, thnh thong lc v quan st

ri ghi nhn xt v tnh cht, cng ca mu v trong sut (v d: vng nht, nu

sm...). Nu tinh du cn vn c v khng trong sut chng t cn tp cht v nc.

II.2.3.1.2 Mi[6,7]

Mi l mt trong nhng biu hin bn ngoi quan trng ca tinh du. Mi mt

loi tinh du c mt mi c trng. Da vo mi c th bit c cht lng v mc

ch s dng ca n.

xc nh mi, nh mt git tinh du ln t giy lc hoc bi mt t vo mu

bn tay ri ngi cch ch c tinh du 20 - 30 mm; c 15 pht ngi 1 ln trong mt gi.

Ghi nhn xt v bn cht v cng mi (thm du, hng, xc...).

II.2.3.1.3 V[6,7]

V cng l mt biu hin bn ngoi quan trng ca tinh du. Mi mt loi tinh

du c mi v ring. xc nh v, dng phng php nm, nm xong ghi nhn xt

bn cht (ngt, ng) v cng ca v (du, thong).

II.2.3.2 Xc nh ch s acid[3,6,7]

Ch s acid biu din bng lng miligam KOH cn thit dng trung ha acid

t do c trong 1 gam tinh du. Bit c ch s acid c th xc nh c lng acid

t do trong tinh du.

Ch s acid ph thuc vo phng php khai thc, mc ti v thi gian bo

qun ca tinh du. Khi bo qun lu ch s acid ca tinh du s tng ln do b oxi ha

v ester trong tinh du b phn gii.

Xc nh ch s acid da vo phn ng trung ha

CHNG II: THC NGHIM

21

RCOOH + KOH RCOOK + H2O

T lng kim trung ha acid t do, tnh c ch s acid.

xc nh cn chnh xc mt lng tinh du khong 2 gam, ha tan bng

ethanol 96, cho vi git phenolphtalein ri chun bng dung dch KOH 0,1N n

khi xut hin mu hng n nh trong 2-3 giy.

Ch s acid c tnh bng cng thc:

m

V5,61IA

=

Trong :

5,61 l lng miligam KOH cha trong 1 ml KOH 0,1N.

V : th tch ml KOH 0,1N dng chun mu th nghim.

m: lng tinh du dng th nghim (g).

II.2.3.3 Xc nh ch s savon ha[3,6,7]

Ch s x phng l s miligam KOH cn thit trung ha tt c cc acid t do

v acid kt hp c trong 1 gam tinh du.

Cch xc nh

Cn chnh xc khong 0,3 gam tinh du vo bnh tam gic 250 mL, cho vo

6 mL KOH 0,1 N trong alcol. Tin hnh song song vi mu khng c du. em 2 bnh

un cch thy qua ng sinh hn, cho si nh trong khong 45 pht tinh du thy

phn hon ton. ngui, thm vo 2 mL nc ct, 2-3 git phenolphthalein, dung

dch chuyn sang mu hng, chun bng HCl 0,1 N trong alcol cho n khi mt

mu hng.

Tin hnh chun mu trng trc khi chun mu tht.

Tnh kt qu: m

)V(V5,61=I 21S

Trong :

1mL KOH 0,1 N tng ng vi 5,61 mg KOH

IS: Ch s x phng

V1 : s mL HCl 0,1N dng chun bnh khng

V2 : s mL HCl 0,1N dng chun bnh cha mu

CHNG II: THC NGHIM

22

m: khi lng tinh du ly lm th nghim.

II.2.3.4 Ch s ester[3,6,7]

L s mg KOH cn thit x phng ho ht lng glycerid c trong mt gam

tinh du, ch s ester l hiu s gia ch s x phng ha v ch s acid. Ch s ester

cng cao th lng glycerin c trong tinh du cng nhiu.

IE = IS IA

II.2.4 Kho st cc yu t nh hng n hm lng v thnh phn tinh

du Bc h

Tin hnh kho st 3 yu t lm nh hng n hm lng v thnh phn tinh

du: thi gian ly trch, nhit v th tch dung mi dng ly trch. Sau khi c cc

iu kin ly trch ti u, ti tin hnh ly trch mt mu nhng iu kin ti u

lm mu so snh thnh phn ha hc vi 3 mu ti u.

Nguyn tc: c nh hai yu t v thay i mt yu t chn c iu kin ti

u cho yu t . Sau thc hin tng t cho 2 yu t cn li. Cui cng tnh phn

trm theo khi lng tinh du thu c v phn tch thnh ha hc bng GC - MS.

II.2.4.1 Kho st thi gian ly trch[4,6,7,8]

C nh 2 yu t nhit v lng dung mi ly trch v thay i thi gian ly trch

nhng giai on 40 pht, 50 pht, 60 pht, 1 gi 15 pht v 1 gi 30 pht. Thi gian

c bt u tnh t lc ta thu c git tinh du u tin ngng t nh xung ng

hng tinh du. Cn v phn tch thnh phn mu tinh du so snh hm lng v

thnh phn tinh du mi thi gian ly trch khc nhau t chn c mc thi gian

ti u nht. Phn tch thnh phn ha hc ca mu thi gian ti u (M1) bng GC

MS.

II.2.4.2 Kho st lng dung mi ly trch[4,6,7,8]

Vi thi gian ly trch ti u chn, tip tc c nh nhit . Ta kho st qu

trnh ly trch tinh du vi nhng th tch dung mi l 200 ml, 300 ml, 400 ml, 500 ml,

600ml. Cn v phn tch thnh phn mu tinh du so snh hm lng v thnh phn

tinh du nhng lng dung mi dng ly trch khc nhau t chn c mc th

tch ti u nht. Phn tch thnh phn ha hc ca mu th tch ti u (M3) bng GC

MS.

CHNG II: THC NGHIM

23

II.2.4.3 Kho st nhit ly trch[4,6,7,8]

Bt bp in trc 30 pht (ty theo nhit tng mi bp un s dng)

nhit cn tin hnh kho st n nh. Cm nhit k vo trong du silicon v lun

theo di nhit ca du. Sau mi tin hnh kho st thi gian v th tch dung

mi ti u va tm c. Ta thc hin ly trch tinh du nhng khong nhit

120C, 130C, 140C, 150C, 160C. Cn v phn tch thnh phn mu tinh du so

snh hm lng v thnh phn tinh du nhng khong nhit ly trch khc nhau t

chn c nhit ti u nht. Phn tch thnh phn ha hc ca mu nhit ti

u (M2) bng GC MS.

II.2.4.4 Kho st s nh hng ca thi gian, nhit v th tch dung

mi n thnh phn ha hc[7]

Tin hnh ly trch mt mu tinh du Bc h kho st (M4) iu kin thi gian

ti u, nhit ti u v lng dung mi ti u. Sau phn tch thnh phn ha hc

bng GC MS v so snh phn trm cc cu t trong tinh du ca mu ny vi 3 mu

thu c mi iu kin ly trch thi gian ti u (M1), nhit ti u (M2) v lng

dung mi ti u (M3). T c kt lun v s nh hng ca 3 yu t thi gian, nhit

v th tch dung mi n thnh phn ha hc ca tinh du Bc h.

II.2.5 Xc nh thnh phn ha hc chnh ca tinh du Bc h bng

phng php sc k ghp khi ph (GC MS) [4,6,7,8]

Tinh du Bc h thu c sau khi ly trch bng phng php chng ct li

cun hi nc c phn tch thnh phn ha hc bng my sc k ghp khi ph

(GC MS) THERMO SCIENTIFIC. Ct s dng trong phn tch l ct c k hiu

TG SQC (15 m x 0,25 mm x 0,25 m), kh mang l heli, vi cc iu kin chy phn

tch nh sau:

II.2.5.1 iu kin sc k

Chng trnh nhit nh sau:

CHNG II: THC NGHIM

24

Hnh 9: Chng trnh nhit

Ram 1:Nhit u 60C, gi 2 pht

Ram 2: tng ln 120C vi tc 5C /pht, gi 1 pht



II.2.5.2 iu kin khi ph

Hnh 10: iu kin chy ca MS

CHNG II: THC NGHIM

25

Ngun ion ha EI:

Nhit bn ph ion: 200C

Nhit stranferline: 230C

Ch qut: Fullscan

Khi qut: 40-400 amu

Tim mu:

Ch tim mu chia dng

T l chia dng: 42

Th tch mu tim: 1 l

Hnh 11: iu kin Injecter

II.2.5.3 Xc nh thnh phn ha hc tinh du Bc h

Thnh phn tinh du c xc nh nh sau:

Xc nh thi gian lu ca cc cht trn GC ging vi thi gian lu ca nhng

cht bit trc.

i chiu ph khi lng thu c vi ph gc trong th vin ph NIST, t

nh danh cc cu t trong tinh du Bc h.

CHNG IV: KT LUN V KIN NGH

26

CHNG III: KT QU V THO LUN

III.1 XC NH CC CH TIU HA L CA TINH DU

BC H

III.1.1 nh gi cm quan

Tinh du Bc h ly trch bng phng php chng ct li cun hi nc thu c

c tnh cht sau:

Hnh 12: Tinh du bc h

Mu: trong sut hi vng nht.

Mi: c mi thm du t nhin

ca cy Bc h.

V: c v cay thm, tnh mt.

III.1.2 Xc nh ch s acid (IA)

Xc nh ch s acid nh phng php trnh by, thc hin o ba ln c kt

qu nh sau:

Bng 6: Kt qu chun xc nh ch s acid

S ln Ln 1 Ln 2 Ln 3 Trung bnh

Khi lng

tinh du (g) 1,0030 1,0032 1,0042 1,0035

Th tch KOH

0,1 N (ml) 0,3200 0,3200 0.3000 0,3133

IA 1,75


27

III.1.3 Xc nh ch s savon ha (IS)

Bng 7: Kt qu chun xc nh ch s savon v ch s ester

Ln 1 Ln 2 Ln 3 Trung bnh

Khi lng

tinh du (g) 1,0034 1,0030 1,0030 1,0031

Th tch HCl

0,1 N chun

mu (ml)

17,6 17,5 17,4 17,5

Th tch HCl

0,1 N chun

mu trng (ml)

14,6 14,7 14,5 14,6

IS 16,22

IE 14,47

V mt ngha ca cc ch s, ch s acid thp cho thy tinh du Bc h thu c

vn m bo cht lng (mi v thnh phn tinh du) ca mt tinh du mi ly trch.

Mt khc, ch s ester ln chng t hm lng ester trong tinh du Bc h nhiu. y

l mt trong nhng cc cu t chnh gp phn to mi thm ca tinh du Bc h nhiu.

Nh vy qu trnh ly trch bng phng php trn t lm thay i tnh cht cng nh

thnh phn ca tinh du.

III.2 KHO ST CC YU T NH HNG N HIU

SUT V THNH PHN TINH DU BC H TRONG QU

TRNH LY TRCH

III.2.1 Thi gian ly trch

Tin hnh kho st thi gian ly trch trn 20 g mu l Bc h kh, thu c kt

qu nh sau:


28

0,0

0,5

1,0

1,5

2,0

2,5

20 30 40 50 60 70 80 90 100

Pht

% tinh du

Bng 8: Kt qu kho st thi gian ly trch tinh du bc h bng phng

php chng ct li cun hi nc

Thi gian ly trch (pht)

Khi lng

mu (g) Dung mi

Th tch dung mi

(ml)

Nhit oC

Khi lng tinh du (g)

Hm lng % tinh du

30 0,2066 1,033

40 0,3146 1,573

50 0,3770 1,885

60 0,4715 2,358

75 0,3755 1,878

90

20

Nc ct

500

120

0,3746 1,873

T kt qu bng s liu trn, tin hnh xy dng th biu th mi tng quan

gia thi gian ly trch tinh du Bc h v s thay i hm lng phn trm theo khi

lng tinh du Bc h thu c tng giai on kho st.

Hnh 13: th biu din thi gian ly trch theo hm lng tinh du Bc h

thu c

Nhn xt: Qua qu trnh kho st v phn tch s liu thu c, cho thy rng: khi

thi gian ly trch tng th hm lng phn trm theo khi lng ca tinh du ly trch

cng tng. V nu thi gian ly trch lu hn 60 pht, nhn thy rng hm lng tinh

du gim li. Nh vy hm lng tinh du t cao nht l 2,358% thi gian 60 pht.


29

Cho nn em chn thi gian ny lm thi gian ti u v c nh thi gian 60 pht cho

cc kho st tip theo.

III.2.2 Kho st nhit ly trch

Tin hnh kho st ly trch 20 g mu Bc h kh nhng khon nhit khc

nhau thu c kt qu nh sau:

Bng 9: Kt qu kho st nhit ly trch tinh du bc h bng phng

php chng ct li cun hi nc

Thi gian ly trch (pht)

Khi lng

mu (g)

Dung mi

Th tch dung

mi (ml)

Nhit oC

Khi lng tinh du

(g)

Hm lng % tinh du

110 0,2991 1,4955

120 0,3082 1,541

130 0,3602 1,801

140 0,3277 1,6385

150 0,2629 1,3145

60

20

Nc ct

500

160 0,2601 1,3005

T kt qu bng s liu trn, tin hnh xy dng th biu th mi tng quan

gia nhit ly trch tinh du Bc h v s thay i hm lng phn trm theo khi

lng tinh du Bc h thu c tng giai on kho st.


30

% tinh du

0,0

0,2

0,4

0,6

0,8

1,0

1,2

1,4

1,6

1,8

2,0

100 110 120 130 140 150 160 170

nhit

Hnh 14: th biu din nhit ly trch theo hm lng tinh du Bc h thu

c

Nhn xt: th trn cho thy rng: hm lng phn trm theo khi lng ca

tinh du ly trch tng khi tng nhit ly trch v hm lng t cao nht l 1,801%

nhit l 130 oC. Nhn thy rng nu tip tc tng nhit cao hn na th hm

lng tinh du t l nghch vi nhit . Cho nn em chn nhit ny lm nhit

ti u v c nh nhit 130 oC cho kho st tip theo.

III.2.3 Kho st lng dung mi ly trch

Tin hnh kho st ly trch 20 g mu Bc h kh vi nhng th tch nc ct khc

nhau thu c kt qu nh sau:

Bng 10: Kt qu kho st th tch dung mi ly trch tinh du bc h bng

phng php chng ct li cun hi nc

Thi gian ly trch

(pht)

Khi lng

mu (g)

Dung mi

Th tch dung mi

(ml)

Nhit oC

Khi lng tinh du

(g)

Hm lng % tinh du

200 0,2095 1,0475

300 0,3300 1,6500

400 0,3618 1,8090

60

20

Nc ct

500

130

0,2986 1,4930


31

% tinh du

0,0

0,2

0,4

0,6

0,8

1,0

1,2

1,4

1,6

1,8

2,0

100 200 300 400 500 600 700 800

ml

600 0,2748 1,3740

700 0,2664 1,3320

T bng s liu trn, ta xy dng th biu din quan h gia th tch nc ct

dng ly trch tinh du Bc h v s thay i hm lng phn trm theo khi lng tinh

du Bc h thu c tng giai on kho st nh sau:

Hnh 15: th biu din th tch dung mi ly trch theo hm lng tinh du

Bc h thu c

Nhn xt: th cho thy khi tng th tch nc ct dng ly trch tinh du th

hm lng phn trm tinh du tng t l thun. V th tch nc ti a thu c

hm lng tinh du cao nht 1,809% l 400 ml. Nu thm nc tip tc nhn thy rng

hm lng tinh du khng tng na m gim li. Cho nn em chn th tch ny lm

th tch ti u.

III.1.4 Kho st s nh hng ca thi gian, nhit v th tch dung

mi n thnh phn ha hc tinh du Bc h

Tin hnh phn tch thnh phn ha hc ca 4 mu: mu kho st thi gian ti u

M1, mu kho st nhit ti u M2, mu kho st th tch ti u M3, mu kho st

c ba iu kin ti u M4. Thu c kt qu so snh nh sau:


32

Bng 11: Bng so snh thnh phn ha hc

Phn trm khi lng % Thi gian lu

Thnh phn ha hc M1 M2 M3 M4

2,29 -Pinene 0,02 0,62 0,53

2,86 -Pinene 0,04 1,05 0,97

3,08 -Myrcene

0,29 0,24

3,21 3-Octanol 0,04 0,05 0,69 0,68

3,7 Limonene 0,1 0,14 1,7 1,55

5,13 -Linalool 0,09 0,14 0,1

6,08 Isopulegol 0,2 0,25 1,25 1,21

6,25 trans-Menthone 9,32 9,43 14,37 16,23

6,76 (-)-Menthol 52,05 47,01 53,77 55,17

7,14 -Terpineol 0,09 0,49

8,04 3,4,4-Trimethyl-5-oxo-

2-hexenoic acid 0,06

8,27 Pulegone 0,2 0,24 7,36 7,15

8,63 Piperitone (p-Menth-1-

en-3-one) 7,99 9,38 2,53 2,26

9,62 Menthyl acetate 18,37 18,17 1,83 1,45

11,7 -Bourbonene 0,06 0,03

12,29 Longifolene 0,03

12,47 -Caryophyllene 1,16 1,67 0,74 0,28

13,59 Germacrene D 0,16 0,26 0,27

14 4-Chloro-2,3-dimethyl-1,3-

hexadiene 0,26 0,53

15,11 (-)-Calamenene 0,17 0,2 0,06

15,42 -Muurolene 0,03 0,02

16,39 Caryophyllene oxide 0,07 0,11 0,03

17,14 -Cedrene 0,1

17,92 -Cadinol 0,05


33

Nhn xt: T bng so snh s liu phn trm cc cht trong tinh du Bc h cho

thy 2 yu t thi gian v th tch dung mi s dng ly trch nh hng n thnh

phn tinh du rt t. Ngc li nhit nh hng ng k n thnh phn tinh du

Bc h. Thy r nht l nhng cht nh -Pinene, -Pinene, -Myrcene kh bay hi

nhit thp, cn cc cht -Cedrene, -Cadinol, -Terpineol, -Bourbonene,

Longifolene li d b phn hy nhit cao. Do mu M1 ta ch thu c -

Cedrene, -Cadinol, -Terpineol, -Bourbonene, Longifolene m khng c mt cc

cht -Pinene, -Pinene, -Myrcene.

Cng thc ha hc ca mt s cu t c trong thnh phn tinh du Bc h:

(-)-


34

Hnh 16: Cng thc ca mt s thnh phn chnh trong tinh du Bc h

III.3 XC NH THNH PHN HA HC CA TINH DU

BNG PHNG PHP GC MS

III.3.1 Thnh phn ha hc

Qua phn tch GC cho kt qu ph nh sau:

Hnh 17: Ph phn tch thnh phn tinh du Bc h bng GC - MS

Trans-Menthone

Pulegone

(-)-Menthol

-caryophyllene

Menthyl acetate


35

Kt qu phn tch bng GC MS cho thy peak c cng ln nht c thi

gian lu ti 6,81 pht. Cho thy y l thnh phn chim nhiu nht trong tinh du Bc

h. Qua nhn dng cu trc ha hc bng khi ph xc nh c hp cht c thi

gian lu 6,81 pht l menthol (C10H20O, khi lng phn t l 156). Khi ph ca

phn t menthol phn tch bng GC MS nh sau:

Hnh 18: Khi ph ca menthol

So snh vi khi ph gc ca phn t menthol c cung cp bi th vin ph

NIST MS Search 2.0 nh sau:

(mainlib) Cyclohexanol, 5-methyl-2-(1-methylethyl)-, (1,2,5)-

10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 1700

50

100

15 18

27

3136

39

41

43

45 48 51

53

55

56

61 65

69

71

73 79

8195

97105

109

112 119

123

138

141 155

HO

10 largest peaks: 71 999 | 81 712 | 95 689 | 55 405 | 41 383 | 82 357 | 69 302 | 123 276 | 96 261 | 67 254 |

Hnh 19: Khi ph ca menthol cung cp t th vin ph NIST


36

Nhn xt: Hai khi ph ca menthol u c cc peak tng i ging nhau . T

kt qu trn cho thy: c hai u c peak [M H ]+ l 155. Cng ca ion phn

mnh ln nht peak m/z 71 v cc mnh ln nh m/z 81, 95, 55, 41, 82, 69, 123, 96,

v m/z 67. S hnh thnh cc mnh c th c gii thch nh sau:

Hnh 20: S phn mnh ca phn t menthol

Di y l kt qu tm tt nhng thnh phn ha hc chnh trong tinh du Bc

h c phn tch bng phng php sc k kh ghp khi ph (GC MS).

Bng 12: Thnh phn chnh trong tinh du Bc h

Thi gian lu (pht)

Thnh phn chnh Phn trm

khi lng (%)

2,29 -Pinene 0,53

2,86 -Pinene 0,97

3,08 -Myrcene 0,24

3,21 3-Octanol 0,68

3,7 Limonene 1,55

5,13 -Linalool 0,1

6,08 Isopulegone 1,21

6,25 trans-Menthone 16,23

6,76 (-)-Menthol 55,17

8,27 Pulegone 7,15

8,63 Piperitone 2,26


37

9,62 Menthyl acetate 1,45

12,47 -Caryophyllene 0,28

13,59 Germacrene D 0,27

III.3.2 So snh kt qu nghin cu

bit s thay i thnh phn ha hc chnh trong tinh du Bc h Mentha

arvensis L. cng nh nhm khng nh tnh chnh xc t kt qu phn tch thnh phn

ha hc do em thc hin. Em tin hnh so snh vi nhng kt c nghin cu trc

y (c thc hin trn cng phng php ly trch v phng php phn tch thnh

phn ha hc):

Trong nc: so snh vi thnh phn ha hc ca cng trn loi Bc h

Mentha arvensis L. c trng ti ti phng Ci Kh, Tp Cn Th do tc

gi V Th Ngc M kho st ( ti thc s di s hng dn ca Gs. Ts

NGT L Ngc Thch)

Nc ngoi: so snh vi thnh phn tinh du Bc h Mentha arvensis L.

c trng v nghin cu ti khu vc pha ty Himalaya. Kt qu nh sau:

Bng 13: So snh kt qu nghin cu thnh phn Bc h Mentha arvensis L.

Phn trm khi lng (%) Thnh phn chnh

Nghin cu Trong nc[16] Nc ngoi [15]

-Pinene 0,53 1,10 0,55

-Pinene 0,97 1,11 0,25

-Myrcene 0,24 0,18 0,31

3-Octanol 0,68 0,45 -

Limonene 1,55 2,87 0,67

-Linalool 0,1 - -

Iso-Pulegol 1,21 - -

trans-Menthone 16,23 19,82 5,08

(-)-Menthol 55,17 51,87 79,64

Pulegone 7,15 1,13 0,73

Piperitone 2,26 6,28 1,08

Menthyl acetate 1,45 2,53 1,63

-Caryophyllene 0,28 0,60 -

Germacrene D 0,27 0,67 -

T kt qu trn cho thy thnh phn chnh trong tinh du Bc h trong nc

tng ng nhau v c thnh phn % cc cu t khng chnh lch nhiu. Mc khc,


38

so vi Bc h c trng ngoi nc th tinh du Bc h trong nc c nhiu cu t

hn, nhng cu t chnh trong thnh phn tinh du l menthol li cha t hn. Ngc

li cc thnh phn nh menthone, linalool, -Caryophyllene, piperitone, limonene

trong Bc h Vit Nam li chim nhiu hn. Nguyn nhn c th do chu nh hng

yu t a l, kh hu khc nhau nn nh hng n thnh phn tinh du khc nhau.


39


IV.1 KT LUN

Qua phn tch hin tng v thnh phn tinh du Bc h ly trch bng phng

php chng ct li cun hi nc, em nhn thy:

Tinh du Bc h c:

Mu: trong sut hi vng nht.

Mi: c mi thm du t nhin ca cy Bc h.

V: c v cay thm.

Cc iu kin ti u cho qu trnh phn tch nh sau:

Bng 14: Kt qu iu kin ti u cho qu trnh ly trch

Khi lng (g) Thi gian

(pht)

Th tch dung

mi (ml) Nhit oC

Hm lng

cao nht %

20 60 400 130 1,8

Xc nh c ch s ha hc ca tinh du Bc h nh sau:

Bng 15: Cc ch s ha hc ca tinh du Bc h

Ch s IA IS IE

Gi tr 1,75 16,22 14,47

xc nh c thnh phn ha hc chnh trong tinh du Bc h Mentha

arvensis L. gm: (-)-menthol 55,17%, trans-menthone 16,23%, pulegone 7,15%,

piperitone 2,26%, limonene 1,55%, menthylacetate 1,45%...

IV.2 KIN NGH

Do hn ch v thi gian nghin cu cng nh trang thit bnn ti cha i

su v cha pht huy ht ngha nghin cu. Trn kinh nghim , em c mt s kin

ngh nh sau:

Tin hnh nghin cu trn mu l Bc h ti v thn cy Bc h ng thi kho

st trn nhiu loi ging Bc h khc nhau.

Tip tc nghin cu ly trch tinh du Bc h bng phng php chng ct hi

nc c s h tr vi sng v phng php CO2 lng.

Kho st qui trnh c lp menthol t tinh du Bc h.


40

Nghin cu chuyn su v th nghim hot tnh khng oxi ha v khng khun

trn cc mng tinh du Bc h.

t tm ra phng php ly trch tinh du hiu qu nht m vn gi c mi

thm t nhin ca tinh du Bc h. Bn cnh cng cho thy tm quan trng ca tinh

du Bc h trong thc t.

TI LIU THAM KHO

PH LC PH GC MS

PH GC MS CA TINH DU BC H LY TRCH THI GIAN TI U

(M1)

Trans-Menthone

Menthyl acetate

Piperitone

(-)-Menthol

-caryophyllene

TI LIU THAM KHO

PH GC MS CA TINH DU BC H LY TRCH LNG DUNG MI

TI U (M3 )

Trans-Menthone

Pulegone

Piperitone

(-)-Menthol

-caryophyllene

TI LIU THAM KHO

PH GC MS CA TINH DU BC H LY TRCH NHIT TI U

(M2)

Trans-Menthone Piperitone

(-)-Menthol

-caryophyllene

Menthyl acetate

TI LIU THAM KHO

PH GC MS CA TINH DU BC MU M4

Menthyl acetate

Pulegone

Trans-Menthone

(-)-Menthol

TI LIU THAM KHO

TI LIU THAM KHO

[1] Raymond M. Harley, Sandy Atkins, Andrey L. Budantsev, Philip D. Cantino,

Barry J. Conn, Rene J. Grayer, Madeline M. Harley, Rogier P.J. de Kok,

Tatyana V. Krestovskaja, Ramn Morales, Alan J. Paton and P. Olof Ryding

(2004), The Families and Genera of Vascular Plants volume VII, Springer-Verlag:

Berlin; Heidelberg, Germany

[2] Tt Li (1995), Nhng cy thuc v v thuc Vit Nam, Nh xut bn Khoa

hc K thut, 595-598.

[3] Huy Bch, Bi Xun Chng, ng Quang Chung,(2003), Nhng cy

thuc v ng vt lm thuc Vit Nam, Nh xut bn Khoa hc K thut H Ni.

[4] Tt Li (1985), Tinh du Vit Nam, Nh xut bn Y hc TP H Ch Minh.

[5] Tiu Chun Vit Nam (TCVN 189:1993), Tinh Du Phng Php Th.

[6] Nguyn Kim Phi Phng (2007), Phng php c lp hp cht hu c, NXB i

hc Quc gia TP H Ch Minh.

[7] Nguyn Th Bch Thuyn, Nguyn Th Diu Thy, Chu Th Thy Hng

(2012), Kho st thnh phn ha hc v hot tnh khng vi sinh vt ca tinh du Hng

Chanh, Trng i hc Cn Th, Tp ch Khoa hc 2012:21a 144-147.

[8] Nguyn Thanh Hu, Trnh Minh Khang, Nguyn Tn Hong Sn, Nguyn Th

Bch Thuyn (2012), Kho st thnh phn ha hc v hot tnh khng vi sinh vt ca

tinh du gng (Zingiber officinale roscoe) v tinh du tiu (Piper nigrumL.), Trng

i hc Cn Th, Tp ch Khoa hc 2012:21a 139-143.

[9] Nguyn Kim Phi Phng (2007), Khi ph, NXB i hc Quc gia TP H Ch Minh.

[10] Asian Journal of Pharmaceutical and Clinical Research (2009), A review on

peppermint oil Shrivastava Alankar, volume 2, Issue 2, April June, 2009, 187-193.

[11] A. Sstrikov, I. alamon (2004), Essential oil of peppermint (Mentha piperita

L.) from fields in Eastern Slovakia, HORT. SCL (PRAGUE), 31, 2004, 31-36.

[12] Ajit K. Shasany, Suman P.S. Khanuja, Sunita Dhawan, Sushil Kumar (2000),

Positive correlation between menthol content and in vitro menthol tolerance in

Mentha arvensis L. cultivars, J. Biosci. Vol.25, No.3, Sep.2000, 263-266.

TI LIU THAM KHO

[13] Gkalp can, Nee Kirimer, Mine Krkcolu, K. Hsn Can Baer and Fatih

demirci (2002), Antimicrobial screening of Mentha piperita essential oil, J. Agric.

Food Chem. 2002, 50, 3943-3946.

[14] James N. Parker, M.D. and Philip M. Parker, Ph.D., (2004), Menthol, A

Medical Dictionary, Bibliography and Annotated Research Guide to Internet

references.

[15] R.S, Verma, L. Rahman, R.K. Verma, A. Chauhan, A.K. Yadav, A. Singh

(2010), Essential Oil Composition of Menthol Mint (Mentha arvensis) and Peppermint

(Mentha piperita) Cultivars at Different Stages of Plant Growth from Kumaon Region

of Western Himalya, Open Access Journal of Medicinal and Aromatic Plant Vol. 1(1):

13-18.

[16] V Th Ngc M (1999), Kho st tinh du mt s loi cy ging Mentha trng

min nam Vit Nam h hoa mi (Lamiacae), i hc Khoa hc T nhin Tp HCM.
LI CAM OANPHIU NG K TI LUN VN TT NGHIPNHN XT V NH GI CA CN B HNG DNNHN XT V NH GI CA CN B PHN BINLI CM NMC LC------(((------PH LC HNHPH LC BNGDANH SCH CC T VIT TTLI M UCHNG ITNG QUANI.1 GII THIU H LAMIACEAEI.1.1 Phn loi hc[1,2]Hnh 1: V tr trong phn loi thc vt ca Bc h Mentha arvensis L.I.1.2 M t[1,2]I.1.3 Phn b v thu hi[1,2]I.1.4 Cng dng ca Bc hI.2 TINH DUI.2.1 Khi qut v tinh duI.2.2 Qu trnh tch ly[3,4,5]I.2.3 Tinh du Bc h[3]I.3 TNH HNH NGHIN CU TINH DU BC HI.3.1 Kho st hot tnh khng vi sinh vt ca tinh du Bc h Mentha piperita L.[13]I.3.2 Thnh phn tinh du ca Bc h Mentha arvensis L. v Bc h Mentha piperita L. nhng giai on sinh trng khc nhau trong vng Kumaon thuc pha ty Himalaya.I.3.3 Tch v tng hp menthol I.4 MT S PHNG PHP LY TRCH TINH DU BC HI.4.1 Phng php c hc[5,6]I.4.1.1 Vt pI.4.1.2 No xtI.4.1.3 p I.4.2 Phng php dng dung mi ha tan[5,6]I.4.3 Phng php chng ct li cun hi nc[4,5,6]I.4.3.1 Phng php c inI.4.3.2 Phng php ly trch kt hp vi sng[4,6]I.4.4 Phng php chit Soxhlet[5]CHNG II: THC NGHIMII.1 A IM, THI GIAN, THIT B, NGUYN LIU V HA CHT II.1.1 a im v thi gianII.1.2 Thit b, nguyn liu, dng c v ha chtII.1.2.1 Thit b v dng cII.1.2.2 Nguyn liu v ha chtII.2 PHNG PHP NGHIN CUII.2.1 X l nguyn liuII.2.2 Ly trch tinh du bng phng php li cun hi nc[7]II.2.3 Xc nh mt s ch tiu ha l ca tinh du Bc hII.2.3.1 nh gi cm quan[6,7]II.2.3.1.1 Mu sc v trong sut[6,7]II.2.3.1.2 Mi[6,7]II.2.3.1.3 V[6,7]II.2.3.2 Xc nh ch s acid[3,6,7]II.2.3.3 Xc nh ch s savon ha[3,6,7]II.2.3.4 Ch s ester[3,6,7]II.2.4 Kho st cc yu t nh hng n hm lng v thnh phn tinh du Bc hII.2.4.1 Kho st thi gian ly trch[4,6,7,8]II.2.4.2 Kho st lng dung mi ly trch[4,6,7,8]II.2.4.3 Kho st nhit ly trch[4,6,7,8]II.2.4.4 Kho st s nh hng ca thi gian, nhit v th tch dung mi n thnh phn ha hc[7]II.2.5 Xc nh thnh phn ha hc chnh ca tinh du Bc h bng phng php sc k ghp khi ph (GC MS) [4,6,7,8]II.2.5.1 iu kin sc kII.2.5.2 iu kin khi ph II.2.5.3 Xc nh thnh phn ha hc tinh du Bc hCHNG III: KT QU V THO LUNIII.1 XC NH CC CH TIU HA L CA TINH DU BC HIII.1.1 nh gi cm quanIII.1.2 Xc nh ch s acid (IA) III.1.3 Xc nh ch s savon ha (IS)Bng 7: Kt qu chun xc nh ch s savon v ch s esterIII.2 KHO ST CC YU T NH HNG N HIU SUT V THNH PHN TINH DU BC H TRONG QU TRNH LY TRCHIII.2.1 Thi gian ly trchIII.2.2 Kho st nhit ly trchHnh 14: th biu din nhit ly trch theo hm lng tinh du Bc h thu cIII.2.3 Kho st lng dung mi ly trchIII.1.4 Kho st s nh hng ca thi gian, nhit v th tch dung mi n thnh phn ha hc tinh du Bc hHnh 16: Cng thc ca mt s thnh phn chnh trong tinh du Bc hIII.3 XC NH THNH PHN HA HC CA TINH DU BNG PHNG PHP GC MSIII.3.1 Thnh phn ha hcIII.3.2 So snh kt qu nghin cuCHNG IV: KT LUN V KIN NGHIV.1 KT LUNIV.2 KIN NGHPH LC PH GC MSPH GC MS CA TINH DU BC H LY TRCH THI GIAN TI U (M1) PH GC MS CA TINH DU BC H LY TRCH LNG DUNG MI TI U (M3 ) PH GC MS CA TINH DU BC H LY TRCH NHIT TI U (M2)PH GC MS CA TINH DU BC MU M4TI LIU THAM KHO

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