3. Phng php 3.

1 Novoenzymes- Tng hp phn tch Butyl butyrate trong ScCO2 t vic s dng bn phn ng khuy 1. M h thng lm lnh (0oC), h thng nhit (40oC) v h thng my iu nhit gii hn (50oC) trc 30 pht. 2. t 50mg Novoenzymes 435( mt CALB c nh thng mi) trong bn phn ng v ng h thng li mt cch cn thn. 3. M van V2 v ng van V1, V3 v sau m bn cha CO2 np vo h thng phn ng cng p sut nh trong chai. 4. M my HPP np vo phn ng vi s iu p CO2 cho n khi t n p sut 100 bar l p sut t c trong phn ng. Sau , ng van V2 v nhanh chng dng my HPP. 5. M my khuy tc 150 vng / pht. 6. bt u phn ng, m van V1 v them 2ml mt dung dch ng phn t c tinh sch (4.54 M vinyl butyrate v 1- butanol, ring tng phn) t tc chy 2ml/ pht bng s dng ng HPLC (chnh xc 1 pht). Sau , dng ng v ng ngay lp tc van V1. 7. Theo phn ng ny, c th c lng khong thi gian xp x phn ng xy ra. Cn thn m van V3 v thi bt trong hn hp phn ng n khi kim sot c lng hexane (v d: 2ml) t vo trong thau trong khong 3 pht. Sau , ng van V3.Phn ng hon ton xy ra khong 3 gi, t 99.5% nng sut tng hp sn phm. 8. Nu cn thit, thm vo ng bm CO2 cho n khi phn ng t ti p sut mong mun (v d: khong 100 bar). 9. Trn chnh xc 500 vi 500 trong 10 mM dung dch propyl acetate (cht ni chun) v phn tch sc k ca 1 dung dch kt qu. 10.Vic phn tch mu c th c mang s dng cho ct ng bn mao dn Nukol trong sc k kh (15m 0.53mm0.5 ) v u d ion ha ngn la. iu kin sc k phi theo chun: phn t mang (N2) ti 8kPa ( tc tng: 20 ml/ pht), chng trnh nhit : 45oC, 4 pht, 8oC/pht, 133oC,t l tch: 5;1, u d, 220oC. Thi gian duy tr nh l: propyl acetate, 3.2 pht, vinyl butyrate, 4.3 pht; 1-butanol, 6.6 pht; butyl butyrate, 7.7 pht v butyric acid, 13.5 pht.

Hnh 5. H thng lc ngang cho mn hnh ng bng gm.(1) bn nhp liu, (2) mng hnh ng, (3) van an ton, (4) mng cha, (5) kim van.

3.2 CALB t do-Emim+ NTf2-Catalyzed Kinetic Resolution ca rac-1Phenylethanol trong ScCO2 s dng trong phn ng ng gi lin tc. 1. ng kim tra cha 2ml emim+NTf2-, thm 65l c 0.9% dung dch CALB v lc trong my hn hp ti nhit bung my cho n khi dung dch tan hon ton. 2. Cho vo Teledyne Isco 220 SFX (10ml tng th tch) vi si cch nhit thy tinh (2g). Sau , thm vo dung dch enzyme emim+NTf2- lm c si cch nhit thy tinh v cui cng t vo trong thng cha c cha thit b chu p sut cao. 3. M bn cha CO2 iu chnh n p sut mong c (khong 100 bar). 4. Chng trnh h thng Teledyne Isco nh l ng lc qu trnh ti p sut c nh (100 bar) v nhit l 40oC 5. M ng thi hai h thng Teledyne Isco v HPLC bt u qu trnh bng vic trng qua 1 lp dung dch ng phn t trong cht nn tinh sch (4.28 M vinyl propionate v rac-1-phenylethanol) ti tc 0.01ml/pht (42.8mol/ pht cho mi c cht). 6. H thng t ng m van thot, tip tc hn hp phn ng to bt mc d c nh c nhit nghim ngt (1ml/ pht, 60oC) trong s kim sot mt

lng hexane trc khi t trong thau . theo di ng hc phn ng, phn tch cht thay th hexane mi 30 pht. 7. Trn 500l dch chit hexane vi 50l ca 100mM butyl butyrate (cht ni chun) v phn tch sc k ca 1 dung dch kt qu. 8. Vic phn tch mu c th c mang s dng cho GC beta-DEX-120 ct mao dn (30m0.25mm0.25m) v u d ion ha ngn la. iu kin sc k phi theo chun: phn t mang (He) ti 1MPa ( tc tng: 205 ml/ pht), chng trnh nhit : 60oC, 10oC/pht, 130oC,t l tch: 100;1, u d, 300oC. Thi gian duy tr nh l: vinyl propionate, 3.2 pht, propionic acid,6.5 pht; butyl butyrate, 7.3 pht ,R-1-phenylethanol,15.4 pht, S-1phenylethanol,16.0 pht, R-1-phenylethyl propionate, 19.3 pht. 3.3 C nh CALB trn mng hnh ng bng s 1. t mng -alumina ln vt cha v bm nc tinh khit vi tc 20 L/pht v a vo bung nhit loi nc v ra mng. 2. ng s s van kim ti lu thong t ti lc phn hi l 2 bar trong h thng, cho php lm kh nhng l nh. 3. t dung dch gelatin/ polyethyleneimine ln bt nhp liu v bm vi tc 20L/pht v a vo bung nhit.Mt film bn trong b mt mng c hnh thnh bi mng lc thi ngang trong dung dch ti p sut phn hi l 2 bar trong 60 pht.Kt qu dung dch lc c thm vo li bn nhp liu trong sut qu trnh.(Hnh 6) 4. M h thng v cn thn di mng s ra. 5. Cho vo mn s dung dch glutaraldehyde v cho chng phn ng li trong 30 pht bung nhit nhm hot ha nhm amino t do. 6. Ra sch mng vi nc loi b glutaraldehyde 7. Thm vo mng s hot ha vi 15mg/mL dung dch CALB( Novoenzymes 525L) trong 0.1 M dung dch m photphat, pH 7.8 v gi qua m 8oC. 8. Thu hi dung dch enzyme cn li v ra sch mng enzymatic 2 ln vi 0.1 M dung dch m photphat, pH 7.8.Tt c enzyme loi b c th s dng xc nh s lng c cht c nh. 9. Gi mng enzymeatic di iu kin kh c P2O5 8oC.

3.4 C nh CALB- Catalyzed Butyl Butyrate Sythesis in ScCO2 s dng mng phn ng ti lu thng. 1. M h thng lm lnh (0oC), h thng nhit (40oC) v h thng my iu nhit gii hn (50oC) trc 30 pht. 2. t mng s enzyme ln thng cha v cn thn ng h thng. 3. M van V1 v V5 ng van V2, V3 ,V4 v sau m bn cha CO2 np vo h thng phn ng cng p sut nh trong chai.

4. M my HPP np vo phn ng vi s iu p CO2 cho n khi t n p sut 90 bar l p sut t c trong phn ng. Sau , ng van V1 v nhanh chng dng my HPP. 5. M bm ti lu thng v thit b o lu lng kim tra lu khi. 6. bt u phn ng, m van V3 v thm 2ml mt dung dch ng phn t c tinh sch (4.54 M vinyl butyrate v 1- butanol, ring tng phn) t tc chy 2ml/ pht bng s dng ng bm HPLC (chnh xc 1 pht). Sau , dng ng v ng ngay lp tc van V3. 7. Theo phn ng ny, c th c lng khong thi gian xp x phn ng xy ra. Cn thn m van V5 v thi bt trong hn hp phn ng n khi kim sot c lng hexane (v d: 2ml) t vo trong thau trong khong 3 pht. Sau , ng van V5.Phn ng hon ton xy ra khong 4 gi, t 99.5% nng sut tng hp sn phm. 8. Nu cn thit, thm vo ng bm CO2 cho n khi phn ng t ti p sut mong mun (v d: khong 100 bar). 9. Trn chnh xc 500 vi 500 trong 10 mM dung dch propyl acetate (cht ni chun) v phn tch sc k ca 1 dung dch kt qu. 10.Mu c th c phn tch bi GC ct bn mao dn Nukol (15m 0.53mm0.5 ) v u d ion ha ngn la. iu kin sc k phi theo chun: phn t mang (N2) ti 8kPa ( tc tng: 20 ml/ pht), chng trnh nhit : 45oC, 4 pht, 8oC/pht, 133oC,t l tch: 5;1, u d, 220oC. Thi gian duy tr nh l: propyl acetate, 3.2 pht, vinyl butyrate, 4.3 pht; 1butanol, 6.6 pht; butyl butyrate, 7.7 pht v butyric acid, 13.5 pht. 4. Ch 1. Vic tinh sch CO2 nn t t nht 99.99%.Thng CO2 c lm kh bi h thng lc c bit. 2. CO2 l kh gy ngt v v vy bn cha phi t ni thng gi tt. 3. CO2 nn cha trong tube thp khng gh. 4. Nhit nghim ngt th cn thit ngn nga tube thot ra t vic ng lnh nh mu ly.Trong trng hp ny, 1/16 tube thp khng gh HPLC t trong thau nhit tnh c th c s dng.Mt my iu chnh lc phn hi hoc hai h thng van kim phi c t iu chnh ton b dng ra.

5. p sut trong bn cha CO2 l 55 bar. iu chnh p sut CO2 trn nh bn bng heli cho n khi t p sut mong mun.iu ny cn thit cho s dng mt thit b vi h thng ng truyn bt CO2 t y ca chai.Vic s dng bn cha CO2 loi ny th c khuyn khch khi ng bm chu p sut cao thng t n iu kin nghim ngt. 6. H thng thi ngang s tip tc xy dng trong tube thp khng gh h tr ln n 30L/pht dng ti lu thng m khng c bt k h thng phn lc. 7. Van an ton phi gm c h thng m trc khi nh c n t n p sut mong mun. 8. Cn thn ng phn ng trnh bt k s mt ha cht hay CO2 khi m h thng c iu p 9. HPP ch c th bm dang lng (nh CO2 0oC v 55bar), khng c dng kh 10. trnh vic tht bi ng van p sut V2 trc khi dng bm HPP. 11.Nu p sut thot ra vt qu gii hn mong mun, van an ton V4 t ng m gii phng lng CO2. 12. trnh mt enzyme c nh v ngn nga kim van kha ngc tr li bi phn t rn, mng lc thp khng gh phi t u tube ly mu (bn trong phn ng). 13.H thng t n trng thi cn bng n nh to ra sn phm 120 pht, t 60% hiu sut sn phm chn la i phn. 14.Thay vo , thit b cm bin nhit c th t ti van V2 ly mu trc tip t mch.