???(???B2)

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???(???B2)
???????????
????????, ?????,????
2
??????????
molecular fluorescence analysis
??? ???????
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?????
?????
????????? ??? ?????
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?????
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??
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??
?????????,?????? ??????,????????? ?????????????,??
????
??????????????? ??????????????? ???
??????????????? ??????????????? ???
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??
(1)X????????????????(????????) (2)?????????(??????
???)
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??
(1)????, ????????? 24???? (2)???? (3)??????????
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???????
?????????? ??????????? ???????????
?????????????? ?????????????
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?????????
1. ???????
S1
S0
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2. ???????
???????????????,????????????,??,???????????????
???????
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3.?????????????
????
?????
??
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internal conversion
internal conversion
vibrational relaxation
S2
intersystem crossing
S1
T2
T1
energy
fluorescence
phsophorescence
external conversion

absorption
vibrational relaxation
S0
l 3
l ?2
l 2
l 1
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??????????? excitation spectrum and
fluore-scence spectrum
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????
????
????
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(No Transcript)
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2.??????
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3.???????
(1)??????????????
(2)????????????
(3)????
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internal conversion
internal conversion
vibrational relaxation
S2
intersystem crossing
S1
T2
T1
energy
fluorescence
phsophorescence
external conversion

absorption
vibrational relaxation
S0
l 3
l ?2
l 2
l 1
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?????????????? relation between fluorescence
and molecular structure
1.????????? ????????????? (1)??????? (2)??????
?????? ??????(?)
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2.?????????
(1)????? ? ?
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(2) ????
??? ?/nm
? 0.07 283
? 0.29 321
? 0.46 400
????,p????????,??????,????????
???
???
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(3)??????
??????????????,????????????????????,????????????
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(4) ?????
??? ??????
? 10
?? 18
?? 20
??? 3
??? 0
? ??????OH, OR, NH2, CN, NR2?,?????
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?? ????????????
??Lambert-Beer??
??,
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???????????????????????????,?
?abc0.05(?????)?,
?????????,????????,
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????????????? relation between fluorescence and
molecular structure
1.????? 2.????? 3. ??pH??? 4.???????
pHlt2 ??? 712 ????
gt13 ???
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?????????
(1)???? (2)???? (3)????????
(4)???????? (5)??????????
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5.???(scattering light)
(1)??(Reyleigh)???
(2)??(Raman)???
??? ???
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?????????instrument of fluorescence analysis
????????? ??????
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?????????

• ????(light source)
• ???(???)(absorption cell)
• ???(monochromator )
• ???(detecor)
• ????????(display)

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????

• ???? ?????-????????,
• ??????????????????.

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????
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????
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???
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??-??????????(???)
??????? ???
It
It
IF,p
I0
I0
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???

• ?????????

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?? ??
?? ???? ???????????
?? a.?????,???,???b.????? ?????b.??????,??????,????????c.?????????,?????????d.??????,?????
????????????? ????????????? ?????????????
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???

• ?????????, ???????????

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?????
??
??
???
?????
????????????-?????????????
???? ????
?????
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??-???????
??(??)?????
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??????
1. ?????(fluorometer)
2. ???????(spectrofluorometer)
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????? ???????
?? ?????? ??
??? ?????? ??
?? ???? ?????????
?? ???? ???????
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??????????? ???4500?????????
???????????????????(?)????
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???????????? fluorescence analysis and
application
?? (1)???? ???-????????24????
????0.10.1?g/cm-3 ?????0.05mol/L
???????????? (2)???? ??????????,?????????? (
3)???? ????????
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??????
????????????????????,????????????????
???????????????????????,??????
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??????
1.?????
F-F0
Fx -F0
cx
c
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2. ?????
??????????????,?????,????????????????,??????
??????,????
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??????????

• (?)??(???)
• 1. ??????????????????????,??????ex287.0nm,?em368
  .0nm?
• 2. ??? ?ex360nm ,?em420nm
• 3. ??? ?ex261nm , ?em284nm

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(?)??(??)

• 1. ???????????????
• 2. ????????????,??????em(3851)nm,?ex301nm??????
  ex(3281)nm,?em425nm

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(?)????

• 1. ????????????????????????????????,??????????????
  ?????,?ex446nm?em506nm?
• 2. ???????,?????????????????????

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(?)???

• 1. ???????????(GPX)???.???(2-Se-CD)???????????????
  ???????(NIH3T3)?????????,??????????????????????
• 2. ??????????????????????-??????????????,?????HPLC
  -????????????????,????? ,????2,3-????4??-?????????
  ????????,????????ex350nm,?em390nm ,

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(?)?????
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1. ?????????????
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2. ????-????
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3. ?????????
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4.????????
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??????????
1928?????????????.????? 1.???. 2.???? 3.???
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• 1. ?????5.2µs1ps,????0.5ps.
• 2.?????? 30000nm/min
• 3.????????????????????
• 4.?????????
• ??????(???)?????????????????

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MFS?UV/Vis ???
?? UV/Vis ?? MFS ??
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??? ??????? molecular phosphorescence
analysis
????? 15 ?????(????????????) 1944 ?
Lewis ???????????? 1957 ? Keirs
???????????????????? 1963 ????????????????????
????
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internal conversion
internal conversion
vibrational relaxation
S2
intersystem crossing
S1
T2
T1
energy
phsophorescence
absorption
S0
l 3
l ?2
l 2
l 1
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??
(1)??????????? (2)??????????
(3)?????????(?I?Br?)??,????-????????,??????????,
????????????,??????????????????????
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????

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

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????(room temperature phosphorescence)
?????????(solid-surface RTP)
?????????(micelle stabilized RTP)
??????????(cyclodextrin induced RTP)
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?????????(solid-surface RTP)

• ??
• (1)????????
• (2)????????????
• (3)??????????????????????
• (4)?????????
• (5)????????,?????????

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?????????(micelle stabilized RTP)
?? (1)???????
(2)????????,??????????????????????? (3)?????
(4)??????????
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??????????(cyclodextrin induced RTP)
?? (1)????????
(2)????????,???????? (3)?????? (4)?????
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???????
1.?????? ?????????????????????????????(????
) 2.???????????????? ??????????,2,4-D???
???0.01 ?g/cm-3 3.?????????
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????

• ??????????????????????,2000
• ?????????????????????????????,1998
• ?????????????????????????????,1998

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