High performance liquid chromatography

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?????????
High performance liquid chromatography
????????
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????????????
?????????? ?????????????????????????
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?????????
1.???
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2.????
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• (1) ?????
• A???????,??150350105 Pa?
• B???????????????????(lt10µm), ???????????,????????
  ,????????????????????
• C??????????????????????????

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(2)??????
??? ?????????,?????????????????????
??,?????????
??? ?????, ???????,??????????????????????
??????
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(3) ????
???????????, ???????????????,
???????
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(4) ?????
?????????,??16 mm,??540
cm?????????????????????
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(5) ???????
a. ????? ????,?????????????
?? ???? ????? ????????(1mm
10mm ,?? 8µL) ???????????????
????,???? ????????
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b. ??????????
??????????
1024??????,???????,???????,??????,?????
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??????????
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c. ??????(Differential refractive index detector)
????????????????
??????????????????????????????????
??????(?????????????)
??????????????????? ?????????????
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???????
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d. ?????(Fluorescence detector)
????????? ?????,???B??????????????
??????????????????
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?????????

• ?? ?-????? (Liquid-solid adsorption
  chromatography)

?????????,????????,??????510µm??????
??????????????????
???????????????????? ???????????????????,?
??????????????????? ????????????????
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?? ?-?????liquid- liquid partition chromatography
???????????(????)
?????????????????? ???????????,???????
?,???????????????(?? normal phase),??,????????????
??(?? reverse phase)?????????????
??????????,?????,????
???????(??????????????????(??)?????????C-18?(???)
?
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?? ??????(Ion-exchange chromatography)
??????????????????????
????????????????,????????????????????,???????
?????????????????????????????????????????
??????????????????,????? ?????RSO3H
M RSO3 M H ?????RNR4OH X-
RNR4 X OH- ????????????,??????
??
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?? ????(Ion chromatography)
??????20??70?????????????,????????????
????????????????????????????,????????????????,????
??????????
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?? ?????(Ion pair chromatography)
?????(???)????????????(???????)??????????
????????????????,?????????????
????????????,????????????????????????
?????????????,???????????
????????????????(C-18?),?????Y???-????-??????,??
??X-??????,????????Y X -????????
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?? ??????(Size- exclusion chromatography)
?????(??????????)
??????????????????????,?????,???????????
???????,?????????????,?????????,???????
????????? ?????????100-105????????????
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?? ????(AC)( Affinity chromatograph)
??????????????????????????,????????
?????????????????????????(??????),??????(???
??),???????????????????????????????????????????
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????????????
????????? Stationary phases of LC
1. ?-???????????? (1)??????
????????????????????????100µm????,???????,???????
? ???10µm??????,?????????
(2)??????? (???????)
3040µm?????,?????????1 2µm??????
????,????
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(3)???????

• ??????? ???????????????
• a. ????? SiOC
• b. ??????SiOSi C
• ??,???????????,????
• c. ???? SiC
• d. ???? SiN

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??????????
(1)???,??????,??????????? (2)???,????,??????,????
?? ???????????,??
(4)????,????????,????? (5)???????
????????? (??????????????) ???????? ??
??????
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2.?-????????
????????????????? ????????????
??510 µm
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3.???????????
???? (1)?????????
????????,????1??????? (2)?????????
????????????(????????) ???? (1)
???????(???????) (2) ???????(???????)
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4. ?????????
(1)???? ??????????????????
???????????????? (2)?????
???-??????????,???? ???????????,???????????
?? (3)???? ??????????
?????????????????,????????,??????????
????????,?????????????
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?????????? mobile phases of LC
1. ????? (1)??????????????,???????????,???
?,??????????? (2)???????????????,???????????
????,?????????,????????? (3)???????????????,?
????????,??????????????????????????????
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2. ?????
??????????????
?????????????? ??????????????????
???? ????????????????????????
??????????????????????????????????,?????????????
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3. ?????
??????,??????????????
?????-????????????????,??????????,??????,?????
?????????,???????????,????????
????????? ?(??) gt ???gt ??gt ??gt ??gt ??gt
??gt ????gt ????gt ???gt ???gt ????gt ??gt ???gt
????gt??gt???gt?gt????gt????gt???gt??gt??(??)
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4. ??????????????
(1)?????????????,?????????????????????????
(2)??????????????????????????????????????????????
??
(3)???????????????,?????????????
(4)?????????????????????????,???????????
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???????????????
????????? factors influenced separation

• 1. ???????????????
• ????????, ????????????????,????????????B/U??
  ,??????,?
• H A C u
• ?????H-u????????????,?????

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• ????????????,?????????,?????????????????
• ?????,?????????????

• ?????,?????????????????
• ???????????????????????

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2.??
????0.5 cm/s?, Hu?????????????????,????
????????????,?????????????????????????
3.??????? ?????????????????????,??????
????????????????,???????????????????,?????????
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???????? choice of separation types
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?? HPLC??? application of HPLC
1. ????????????? ????????(37
50?m) ?????? ? ?1.5
mL/min ???50cm?2.5mm(??)
??????????
???????????????????
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2. ???????
????????????? ???20??-? 100??
??????,2/min ? ?1mL/min ? ?50 ºC
? ?70 ?104 Pa ????????
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????????? preparative liquid chromatography
??????(???)?????? ????(??8mm,??15
30cm),?????0.11mg
1.???????(???) ????????????????
??????????????????? ?????????????????,???
? ?????????,????????????k??10???
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2. ?????????
?????,??????? (1)???????,??
?????,?????? (2)??????????
??,???????????????(??)?,???????
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3. ???????
???????????????,??????????????????????
?? ?????2050mm,??50cm?