methodology ir raman spectroscopies

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ΦΑΣΜΑΤΟΣΚΟΠΙΑ ΥΠΕΡΕΡΥΘΡΟΥ (IR SPECTROSCOPY) ΦΑΣΜΑΤΟΣΚΟΠIΑ Raman (Raman SPECTROSCOPY) 1000 1200 1400 1600 1800 1391 1547 1519 1373 1334 1275 1158 1041 1228 1446 1627 1670 1732 2nd derivative Wavenumber (cm -1 ) cA peptide oriented fiber stained with Congo red ATR units ΦΑΣΜΑΤΟΣΚΟΠΙΕΣ ΤΑΛΑΝΤΩΣΗΣ Βιοφυσική Φεβρουάριος 2015 Επικ. Καθ. Βασιλική Οικονομίδου 1

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Methodology IR Raman Spectroscopies

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  • (IR SPECTROSCOPY) I Raman (Raman SPECTROSCOPY)

    1000 1200 1400 1600 1800

    1391

    1547

    1519

    1373

    1334

    1275

    1158

    1041

    1228

    1446

    1627

    1670 17

    32

    2nd

    deriv

    ative

    Wavenumber (cm-1)

    cA peptide oriented fiber stained with Congo red

    ATR

    units

    2015

    . . 1

  • : H

    : EINAI MIA STEIN: /

    2 . . .

  • / .

    3 . . .

  • / : : o

    =c/

    c: (3 x 1010 cm/sec)

    4 . . .

  • cm. :

    = 1/ = /c

    5 . . .

  • / . : E = h

    h: Planck=6,624x10-34 J s

    6 . . .

  • / 3 : A

    7 . . .

  • H

    ~ 4000-8000

    8 . . .

  • ?

    hv1 hv2

    hv3

    hv1

    hv3

    9 . . .

  • :

    : To , Planck: E = h (, ) . . , . 1.

    10 . . .

  • Lambert-Beer

    (: )

    leII = 00I

    l

    I

    11 . . .

  • 2 :

    = / 0 %

    12 . . .

  • A=Absorbance=

    IIA 010log=

    0I

    l

    I

    lCIIA ..log 010 ==

    Lambert-Beer

    (: )

    :

    13 . . .

  • () Boltzmann

    (=0). 1 . .

    14 . . .

  • 2 ( ) m1 m2.

    H ( , ) :

    E= Kx2

    .

    , .

    , .

    ;

    m2 k m1

    15 . . .

  • orse

    1

    16 . . .

  • ;

    m1 m2 .

    m1 m2 k

    H k. , Hooke. : =1/2 k/ (= ) 1/=1/m1+1/m2 =m1m2/(m1+m2) 17

    . . .

  • ()

    : m1 m2 :

    m1>>>>>>>>>>>>>>>>>>>>>m2 m1=m2

    : a. 1/=1/+1/m2=1/m2 . =m2 b. 1/=1/m1+1/m2=2/m2 . =m2/2

    18 . . .

  • !

    , ( , ). Morse , . (-) ( , ).

    19 . . .

  • orse

    1

    20 . . .

  • 21 . . .

  • . :

    . ,

    : 3N-6 3-5 ( )

    22 . . .

  • 23 . . .

  • IR

    :

    , , ...

    in-situ :

    24 . . .

  • ( ) .................................................................................................................. ................................................................................................................. ................................................................................................................

    :

    25 . . .

  • ,

    26 . . .

  • 27 . . .

  • (IR)

    1000 1500 2000 2500 3000 3500 4000

    A

    Wavenumber (cm-1)

    vmax

    28 . . .

  • : Wavenumber = ( )

    v -

    1

    = (cm-1)

    cv =

    29 . . .

  • A=Absorbance=

    IIA 010log=

    leII = 0 0Il

    I

    lCIIA ..log 010 ==

    Lambert-Beer

    (: )

    (: )

    :

    30 . . .

  • 1000 1500 2000 2500 3000 3500 4000

    A

    Wavenumber (cm-1)

    ?

    31 . . .

  • 32 . . .

  • ...

    = 1100 - 1800 cm-1

    1000 1500 2000 2500 3000 3500 4000

    A

    Wavenumber (cm-1) 33 . . .

  • 1200 1400 1600 1800

    1660

    1273 13

    90

    1157 12

    28 1450

    1531

    1628A

    Wavenumber (cm-1)

    1100 - 1800 cm-1

    34 . . .

  • A : 1600-1700 cm-1

    : 1500-1600 cm-1 : 1220-1330 cm-1

    ( )

    35 . . .

  • A : C=O

    : C-N , - : N-H

    36 . . .

  • :

    -

    -

    -

    I

    1645 - 1660

    1620 - 1640

    1662 - 1695

    1645-1655

    II

    1546

    1520, 1530

    III

    1295 - 1328

    1225 - 1250

    1257 - 1288

    IR

    37 . . .

  • 1200 1400 1600 1800

    1660

    1273 13

    90

    1157 12

    28 1450

    1531

    1628

    A

    Wavenumber (cm-1)

    38 . . .

  • 1000 1500 2000 2500 3000 3500 4000

    A

    Wavenumber (cm-1)

    N-H C-H O-H

    3280 cm-1

    -: 3290 - 3300 cm-1 - : 3280 3300 cm-1

    39 . . .

  • 900 1000 1100 1200 1300 1400 1500 1600 1700 1800

    A

    Wavenumber (cm-1)

    1200 1400 1600 1800

    1660

    1273 13

    90

    1157 12

    28 1450

    1531

    1628

    A

    Wavenumber (cm-1)

    40 . . .

  • 1000 1200 1400 1600 1800

    1391

    1547

    1519

    1373

    1334

    1275

    1158

    1041

    1228

    1446

    1627

    1670 17

    32

    2nd

    deriv

    ative

    Wavenumber (cm-1)

    cA peptide oriented fiber stained with Congo red

    ATR

    units

    2

    S : M: W:

    41 . . .

  • 42 . . .

  • , .

    CO2

    : 43

    . . .

  • 44 . . .

  • : M . , , . , , . , Rayleigh.

    45 . . .

  • Raman (1928)

    O C.V. Raman (laser) , . Raman. , , . Rayleigh Raman , (h= h-). Stokes ( ), (h= h+). - Stokes 46

    . . .

  • 47 . . .

  • Raman: , ( Rayleigh) ( Raman, ). ( ) .

    48 . . .

  • H Rayleigh Raman ( Stokes anti-Stokes) .

    Stokes anti-Stokes Boltzman .

    Raman Stokes ( laser )

    RAMAN

    49 . . .

  • 600 800 1000 1200 1400 1600 18000,015

    0,020

    0,025

    0,030

    Ram

    an In

    tens

    itycA-peptide

    925

    8948

    51

    643

    75572

    4

    5975

    23

    1615

    11681

    102

    1032

    1003

    1333

    1241

    1453

    1669

    Wavenumber (cm-1)

    FT-Raman

    50 . . .

  • Raman

    - -

    -

    I 1650-1660 1665-1680 1665, 1690 1660-1667

    A II 1516, 1545 1535, 1560 (I) 1550-1555

    1567

    (II) 1545, 1555

    1560

    A III 1260-1290 1230-1240 1290-1330

    51 . . .

  • K (cm-1) 510 S-S (g-g-g)

    525 S-S (g-g-t)

    540 S-S (t-g-t)

    622 Phe

    643 Tyr / Phe

    759 Trp

    830 Tyr

    850 Tyr

    882 Trp

    1003 Phe C-C

    1035 Phe

    1100-1110 C-N

    1171 Tyr

    1206 Tyr, Phe

    1245 (. -)

    1268 (- ??? -)

    1341 (-)

    1447 CH2

    1553 (-)

    1611 Tyr, Phe, Trp

    1671 I (. -)

    2800-3100 C-H /N-H / OH 52

    . . .

  • 53 . . .

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