NOI DUNG VA PHUONG PHAP NGHIEN CUU
2.1. NOI DUNG NGHIEN CUU
2.1.1. Kluio siit cac dieu kien de xiic dinli lugng vet idt bang phuang phap trac quang-DHXT, dung pluin ling chi (hi [Cé'^-As-^^] va [clorpromazin-HiOj]
Hien niiy đ xac dinh lugng vet idl bang phuang phap phan tfch DHXT ngudi ta dang sir dung nhieu pluin ling chi ihi kluic nhau (biing 1.7, trang 36), mdi loai phan ling chi Ihi iiiiy deu cd uu va nhugc didm nhál dinh. Hai pluin ling chi Ihi [Cc'^^-As^^J va [clorpromiizin-H202] dugc nhieu liic giii nghien cuu vii ap dung đ pluin lich idt trong nhieu đi tugng khiic nhau [51], [63], [124]. Tuy nhien cung tuy thuge vao dieu kien cu Ihd (hda chai, nude cai, Ihiei bi va dung cu) ciia lung phdng ihi nghiem, khi mudn iip dung nhung pluin ling chi thi nao cung can phiii khiio siit ciic diic trung quang hgc va dong hgc ciia liing he pluin ling. Tren ca sd nhung dieu kien da chgn dugc, diinh gia do lap liii, do chinh xiic va gidi hiin phat hien ciia phuang phap trac quang dong hgc xac tiic, he [Ce(IV)-As(III)l va he [clorpromazin-H202]. Cac yeu id Ciin khiio siil đi vdi phuang pluip pluin tfch trac quang DHXT bao gdm:
+ Ndng do ciia Ciic tiic nhan Iham gia phiin ling chi thị
+ Ndng do va loiii axit iinh hudng deii lieii trinh Ihuc hien phiin ung + Anh hudng ciia nhiet do va ihdi giiin piiiin ung deii idc do phiin ung + Ciic ion Ciin va ciic chát kluic iinh hudng deii pluin ling chi tin
2.1.2. Nghien ciiu dieu kien pluin huy nuiu de xiiy dung qui trinh phan tich lugng vei idt trong: nude tieu, sua; tdc, huyet thanh ngudi va nude sinh hoat
Nlurng đi tugng nghien ciiu lien day cd Ihanh phdn nen rál phuc liip, do vay Ciin phiii nghien cuu phuang phiip phan luiy mau thfch hgp cho liing đi tugng đ loiii tru" Ciic thiinh pluin trong nen cd thd iinh hudng deii pluin ung chi thị Mill khac, tuy thuoc Viio ham lugng idt trong tiJiig đi lugng phan tfch, vdi phuang phap DHXT, nhiet do vii thdi gian phiin ling sc iinh hudng niiinh deii tdc do phan ung chi thi, đng thai trong lung đi lugng cung phiii kluio sat mot sd chiit cd the ciin trd hieu ling xiic tac ciia idl len pluin ling chi Ihj. Trong pluin nay Ciln pluii khiio sal nhung ycu id siiu day:
+ Tun dieu kien thfch hgp đ phan hiiy lUc^u cho tirng đi lugng rieng biet + Danh giii do tin cfly ciia qui irhili pluin tfch Xiiy dung dugc qua ciic ycu td nhu do chnih Xiic, do thu hdi do hip hii; so siinh kei qua phan tfch qua hai phuang phiip Xiic dinh khiic nhau
+ Khiio Siit khoiing tuyen tfnh, vii giai hiin phat hien cho lung đi tugng nghien ciiụ
+ Ap dung phan Ifch tren đi lugng lliirc te
2.1.3. Xiic dinh nidi luang quan cua luini lugjig idt trong cac đi lugng: nude lieu, sua mc, toe, huyei thanh dcudng rdn tre sasinh, nude sinh hoat
+ Mdi tuang quan giiia ham lugng idt-nieu ciia hgc sinh lieu hgc (do ludi tir 8 12 ludi) theo viing sinh tluii va Ihdi gian nghien cuụ
+ Mdi iiXOng quiin giua hiim lugng idl-nieu ciia phu niJ miing thai theo vimg
sinh thiii Vii thiii kị
+ Mdi tuang quan giua hiim lugng idl trong nude tidu vdi trong sQa ciia cac ba mc dang cho con bii theo vung sinh thiiị
+ Mdi luang quan giua hiun lugng idt-nieu vdi idt Uong tdc cua ngudi tiirdng thanh, + Mdi tuang quan giua trgng lugng ca ihd vdi luim lugng idl trong huydt thanh d cudng rdn tre sa sinh.
+ Mdi tuang quan giiia ham lugng idt trong nude sinh hoiil theo viing sinh thai + Danh giii hieu quii chuang tiinh phdng chdng CRLTHI d linh ThiíaTliien Hue 2.2. VAT LIEU VA PHUONG PHAP NGHIEN CUU
2.2.1. Thici bj
1. Cdn phdn tich. Gui phiin tfch dien Siiuler (My). Do chfnh xac ciia can ±01 mg.
2. Mdy trdc quang AE-22 (Nhat). Vijiig budc sdng tii 400-760 ± 2 nm.
Miiy cd ihd do ciing mot liic ba mfiu, vdi cuvel thiiy linh cd chieu day 1 cm
3. Mdy trdc quang ANA-172 (Nhat). Miiy sir dung trong khoiing budc sdng
tií 340-900 ± 0,1 nm. Miiy hien sd vdi 3 elm sd sau dau phay, cuvet thuy linh cd chieu diiy 1 cm, do cung mgl liic ba niiiụ Trudc khi do phai chinh ve do iruyen quang 100, hoac do háp ihu 0,000, sau đ mdi lien hiinh do miiụ
4. Mdy trdc quatig Perkin-Ermer (USA). 5. Gamma phd ke hai kenh Oakfield (Atdi)
6. Mdy dieu nhiet cdch thuy (Cole-Palmer) 7. Mdy sieu dm cdch thuy (Cole-Palmer) 8. Ddng hd bam gidy (Anh)
9. Ld nung (Dítc) 10. Mdy ly tdm (Due)
11. Mdy cat nitd'c hai Idn (cai trong thici bi thiich anh), Anh qudc san xuaị
2.2.2. Dung cu va hda chiit
- Ong nghiem cd niit nuii: 18x180mm (hang Sibala, Nhai) - Ong li tam cd ndp (Nhat)
- Chen nung bfmg su, cd nfip diiy (TQ) - Chai nhua trang (hang Sargent, My) - Miropipel (Anh)
Hiiu hei cac hda chai sir dung deu thuge loiii linh khidt phan tich ciia cac hiing Merk, Prolabo, Cedex. Rieng (NIl4)2S20«, Na2S03, H2O2, KCIO3 la loai tinh khiei hda hgc ciia Trung Qudc
'" Núdc cat klwng cd idt.
Dimg nude ciii hai kin, kiem hda bfmg K2COV rdi cai liii trong bg chung cai bang Ihiiy linh. Nude cai nay dugc chiia trong binh polyelylen kfn, dung de pha che ihudc ihiị
Tiii Cii Ciic dung cu ihiiy linh phiii ngam khoiing 12 gid trong dung dich HNÔLl, triing bang nude Ciit, sáy kho 105"C. trudc khi phan tfch phai tiang h i bfmg nude ciii khong cd idt (tir day uii cfi nude ciii khong cd idt dugc diing đ pha thude thií, hoac de pha loang: dugc ggi la nude citl).
'^ Dung dich chudn
Dung dich churln gdc KI: Imgl/ml. trudc khi dem can đ pha che, KI dugc say d I05"C trong 8 gid, đ loiii I2 tu dọ Can 0,654g KI hda tan trong nude cai, rdi
dinh miic den 500 ml. Dung dich luiy chiia irong chai thiiy tinh miiu nau vii kiem
hda bfmg Viii gigt KOH 4M đ liiinh hien lugng I bi háp phu len Ihanh binh.
Dung dich chuan Irung gian Kl: 1 j-igl/ml dugc pha che lir dung dich chudn gdc Ciic dung djch chụ^n lam vice (20, 40, 60, 80, 100, 120 ng/ml,...{ pha loang trong dung djch 0,2M KOH)) dugc pha loang tu dung dich chuan trung gian (chi pha Irong ngay Irudc khi su dung)
Dung dich chudn gdc KIO,: lOOOj-igl/ml. Can 168,5 mg KIÔ, hda lan trong lOOinl nude cai khong cd idl. Dung djch chuaii lOi-igl/ml: pha loang lir dung dich chuiln gdc. Ciic dung dich chuan lam vice: 0,1; 0,2; 0,3; 0,4; 0,5jLigr /lOOml ...dugc pha loang lir dung dich chuaii trung gian: lOj-igl'/ml.
"\ Cdc dung dich khdc
Dung dich axil cloric (IICIỌO, 28%: hda lan 500g KCIÔ irong 900ml HjO
dun cho Idi khi lan hei, them Itr lu 375 ml IICIO4 70% kháy lien luc. D& dung dich
trong ui hmh qua dem, ngay hdm sau chiei liiy dung dich trong. Bao quan dung dich trong chiii nau, de liuih d lỐC, dung dich nay ben khoang 6 thang.
Dung dich iixil iisenil 0,05M: hda lan 9,9 g AS2O3 + 25g NaCl + 200 ml H2SO4 2,5 M + 400 ml nude ciiị Dun soi cho lan heị Dinh miic bang nude cai dii mgl hị Dung dung dich trong chai nau, biio quiin d chd tdị
Dung dich xeiic (IV) iuiioni sunfal 0,075M: hda tan 47,4g Ce(S04)2.2(NH4)2S04.2H20 Irong lift dung dich H2S04l,25M. Dung dung dich trong chai nau, biio quiin a chd idị
Dung dich amoni i^csunfat 1 mol/L: hda lan 22,82g amoni pesunfiU trong 100 ml nude ciii (dugc pha ngiiy klii diing, va dung laii hda chitl siui xuiii Uong nam sir dung)
Qic dung dich axit HCl, H2SO4,1 liro4, HNO3 dugc pha tii dung djch axit dam dac Ciic dung djch clopromazin, KOH, Z11SO4, Na2C03, KBrO,,, NaF, KSCN, K2Cr207, KIO3, S03^",...dugc pha lir linh the tuang ting
Dung dich ciic ion gay ciin: Fê, CÍ, NO2', Câ, K^, W^....6{xac pha tii linh Hid tuang liug
Dung djch H2O2 lOM. Ndng do cua dung dich dugc kidm tra liii bang phuang phiip chuiln do vdi KMiiOj trong moi Irudng axil
2.2.3. Phuong p h a p nghien ciiu 2.2.3.1. Liiy mau
Trong luiin an luiy tai cii ciic nuiu nghiem dugc láy lai linh Tlma lliicn Hue Chgn ba viuig sinh thai đ liiy nuiu: Mien niii gdm mgl sd xa d hai huyen Nam Dong va A Ludị Vung ven bien gdm 4 Xil: Thuiin An, Phii Tliuiln, Phu Hiii, Phii Dien: nam ven bidn Dong Ihugc huyen Phii Vang. Vung đng bfmg gdm mgl xa nam rai rac trong tinh Thua Thien Huẹ Tlidi gian nghien cuu lu lluing 7/1996 deii 12/2000. Lay mSu theo phuang phiip ngSu nhien dan. Mau dugc láy nhd sir trg giiip ciia chuang trinh
phdng chdng CRLTHI va Trung tam y le DiX phdng tinh Tliiia Thien Huẹ
+ Miiu nude tieu dugc láy iren nhung đi lugng sau
^ Hgc sinh tieu hgc {66 ludi lir 8 ludi deii 12)
O mdi xa (hoac phudng) chgn mgl trudng tidu hgc, trong moi trudng chgn ngfiu nhien 40-50 hgc sinh đ liiy nuiu nude tidụ Chiia miu trong chai nhua PE, biio quiin niiiu d nhiel do < 10"C cho deii khi phan tfch miu
^ Phu nu* dang mang thai (ihdi giiin mang thai lir thang thu 2 den thang thu: 9)
Chgn ngau nhien 287 phu nix mang thai d do ludi tu 20 den 35 ludi, sue khde
binh thudng, sinh sdng d ba viing sinh thiii: vung niii (huyen Nam Dong va A Ludi); viing đng bfmg (mot sd phudng xa d ngi va ngoai Ihanh phd Hue); viing bien (mot sd
xa thuoc huyen Phii Viuig) thuge tinh Tliira Tliien Hue 6& phdng van va lay mau nude
tidụ Ddng thdi, Irong mdi vung, chgn khoiing 40-50 phu nii, siic klide binh thudng ciing do ludi nhung kliong cd lluii đ liiy mSu nude tidu lam nhdm đi chung.
^ Ba mc dang cho con bii (tudi ciia Ire bii mc lii mgl thang den sau Ihang) Chgn ngilu nhien 247 cac bii mc diing nuoi con bang sira mc (ludi con tii" mgl deii siiu thiing), tudi mc tii 25 den 35 ludi, cd sue khde bhih thudng, hien dang sinh sdng d ba vung sinh thiii khac nhau ciia linh Thira Thien Hue: mien niii (huyen Nam Dong); viing đng bfmg (gdm cac xa va phudng: TliiiyBieu, Phudng Diic, Tiiy Lgc, vii VI Da thuge thanh phd Hue); vung ven bidn (gdm cac xa: Thuan An, Phii Thuan, Phii Dien thuoc huyen Phii Vang); đ phdng ván va láy mau nghien cuu( nhdm A). Ddng thdi d mdi vung sinh tluii chgn khofing 40 phu nu (ciing do ludi, cd sue klide
binh thudng, khong cd thai vii khong cd con < 5 tudi diing sinh sdng trong cung mgl
nhii vdi ngudi me) 66 liiy mfiu nuac tidu lam nhdm đi chung (nhdm B)
Mill! sua va nude tidu dugc hty đng thdi tren cung mgl ngudi mc vao budi siing, dung mfiu Irong chai nhua PE, bao qufm miiu sua d -20''C , mau nude tidu d < 10"C cho den khi phan tfch nuiu
* Phdng viTn ciic đi tugng nghien cuu ve hien irang sue klide va tinh hinh sir dung cac thuc pham giau idt (dieu ira bfmg phieu dieu tra: trinh bay d phan phu luc)
^ Ngudi trudng thanh (sinh vien dai hgc y Hue, ludi lir 18 dcii 25)
- Ddi vdi mau nude tidu 24 gid: hty loan bg nude tidu (kd ca liic di lieu) lu 6 gid sang honi trudc den 6 gid siing hom sau, do idng the tfch nude tidu, lay vao chai dung mfiu: khoiing 2-3 ml
- Ddi vdi nuiu nude tidu ngilu niiien: hty vao budi siing, thd tfch nude lieu lity nhu lien
Dung mfiu trong chai nhua irdng, biio quan miiu trong lit lanh d < 10"C cho den khi pluin tfch mau
+ Miiu sua mẹ Lity mfiu siia me Iren đi tugng đng thdi vdi mau nude lieu ba me dang cho con bu, mdi mfiu hty khofing 2 ml, bao quan mSu d - 20"C cho den khi plum tfch nuiu . Sua bd, siia hop, sua bgt hty tren thi trudng tinh Tliira Tliien Hue
+ Miiu huyei thanh dcudng rdn tre sasinh. Chgn cac ba me sinh con tiii khoa Siin phu Benh vien Trung uang Hue vii mgl sd benh vien vung phu can đ láy mfiu maụ Miiu mau dugc lity d cudng rdn ciia tre sa sinh viio thai didm ngay sau klii dẹ Láy mau tu mieng cudng rdn cfu ve phfa rdn mc. Lity khofuig 5ml mau, li liim 2500 vdng/phiit, trong 10 phiit; lay huyei thiuih biio quiin d - 20"C cho deii klii phan tfch maụ
+ Miiu luc. Lity tdc d vimg chiim diiu, cdt hty doiin tdc ciich mat da diiu
khoiing iix 1-5 cm. Dung mfiu tdc trong liii nilong va biio quan d 25"C
+ Miiu nudẹ Lay mfiu nude sinh hoiil d ba viing sinh thai: mien niii, vung đng biing, vung ven bien. Dung miiu vao chai nhua PE, mdi miiu hty khoang 200 ml nude, biio quiin nuiu nude d lu liinh cho den khi phan lich.
2.2.3.2. Nghien cuu a p dung mo hinh toiin hgc de qui hoiich hda thuc nghiem nhiim lim dieu kien Ihi ngiiicm tdi uu cua ciic ycu td iinh iiudng den phiin ling chi thi [Cc(lV)-As(lII)] vii clopronuizin-H202] va dieu kien de phan huy mau nude tieu
D6 hiin che nhung nhugc didm ciia phuang phiip dan bien; nhung dieu kien
tdi uu ciia Ciic tiic nhan tham gia phfin ung chi thi va didu kien đ phan hiiy mfiu nude lieu dugc lien hanh theo phuang phap qui hoiich hda thuc nghiem, dua tren vice xay dung mgl mo hinh loiin hgc thfch hgp. Cac mo hinh loiin hgc nay dugc Xiiy dung dua vao viec lam thf nghiem theo ma tran qui hoach hda thuc nghiem bile hai Ulm xoiiỵ Trong đ phuang pluip dudng đc nhai va phuang phiip dao ham rieng dugc diing đ lim nhirng cue trj ciia ham muc lieu mo la cac yeu id anh
hudng Ciin Xiic dinh. So lieu thf nghiem dugc xix If tren chuang trinh Statgraphics.
2.2.3.3. Phuong pluip Xiic djnh idt
Ap dung phuang phap tga, phuang pluip an djnh thdi gian, đ xay dung đ
thi chuiin. Diing đ thi đ 66 xac dinh hiim lugng idt trong mSu phan tfch. Six dung
phuang phiip them chufin đ so siinhvdi phuang phap an dinh thdi gian. Sii dung phiin mem STARGRAPHIC 7.0; ORIGIN đ thici lap phuang trinh dudng chuan theo phuang phiip binh phuang idi thidu va tfnh he sd tuang quan (r). Hiim lugng idl Irong nude tidu dugc bidu didn bfmg j^igl/dl (dl: 100ml)
2.2.3.4. Phuong phiip diinh giii do tin cay ciia quy trinh phan tich
Do tin cay cua quy Irinh phan tfch dugc danh gia qua nhung ycu td nhu do
lap liii, 6<^ thu hdi, do chfnh Xiic, GHPH va khofing tuyeii tfnh.
• Do liip liii dugc diinh giii qua do lech chufin luang đi (luiy he sd bicii dong: CN).
^ Xiic dinh do thu hdi dua vao phuang phap them nhung ndng do idt chuan
vao Ciic mfiu nghiem ma ndng do idt da xac dinh % do thu hdi = (L mfiu do)/(r If ihuyei) x 100
• Gidi hiin pluil hien dugc ifnh iheo "quy ific 3a". Ciic giii tri ciia GHPH duac Ihuc hien tren miiy ifnh nhd vao ciic phiin mem cd sán. Diinh gia do chfnh
Xiic ciia phuang phiip nghien ciiu bfmg ciich xiic dinh ham lugng idt trong mau chuiin Vii so Siinh vdi kei quii phan Ifch vdi phdng ihf nghiem khiic.
2.2.3.5. Xii If sd lieu phiin tich bilng phuang pluip thdng kc [31, [20], [27], [35], [58], [84], [86], 195], [119], [136]
So sanh tri irung binh ciia hai Uip sd lieu bang test Student (test "t"), neu
tap sd lieu phan bd khong chuan, thi phiii chuan hda bang each chuyen sd lieu qua he log ihap phan. Cong thuc tfnh I:
^ A ^ ^ B ( 2 , 1 )
' A " B
Trong đ: x^; Xg; HÂ ^lu '•> ^A '^ ^H '• Íl'^ ^^^^ 1^ hi trung bhili; sd mSu; do
lech chuiin ciia hai tilp sd lieu (trudng hgp n > 30). Khi n < 30 thi ta thay the â va
OQ biing mot phuang sai chung cho hai mau {G ~ â-^ â) vao cong thiic (2.1)
So .sanh tuang quiin giua hai Uip sd lieu dimg phuang trinh tuang quiin hdi qui va he sd luang quan r d Xiic suai p ling vdi do lir do n-2. Cong thuc tfnh r va t nhu sau:
\/Z(x-><)'E(y-y)' ^^-^'
Trong đ: x.x;ỵy l^n litcrt la gia tri rieng le va gia trj trung binh ciia hai tap sd lieu, n hi cd mfiu nghien ciiu, t la xiic suili p d bac tu do n-2.
Phuang Irinh ciia dudng hdi qui giua x vil y nhu sau: y = b + ax
Trong đ: g . ^ ^ y . va ^ ^ Z(x-><)_,; ^ ^ I^.^lzl^. ; b = y - x a
ox ' V n - 1 ^ V n - 1
Tiii Cii Ciic phep ifnh thdng ke deu thuc hien tren may tfnh cii nhan vdi cac phiin mem nhu Origin, excel, Sliigraphic 7.0...
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