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Extraction of 3D Data
XY plane = Chromatogram ; ZY plane = spectrum
Chromatogram
Absorbance
1 2
Time
Absorbance
Spectrum
Wavelength
Dr. Shulamit Levin
Coelution of 2 Peaks
A
Coelution detection at a single wavelength Coelution is the sum of absorbance of 2 peaks A and B
B AU
Elution Time
Dr. Shulamit Levin
t R(1) Rs = t0
w1
t R(2)
tR (2) - t R(1)
1/2
(w 1 + w 2 )
w2
R=0
R=0.3
R=0.7
R>1.0
R=0 Purity Angle not effective; Match Angle useful R=0.3 to R=0.7 Purity & Match Angle useful R>0.7 Match Angle not useful
Dr. Shulamit Levin
Purity verification
Apex
sin j =
( B ij s j Ai ) 2
Library identification
Standard Unknown
i =1
B ij 2
i =1
Absorbance
0 deg 90 deg
Time
Absorbance
0 Sin 1
Time
Time
Peak Purity analyzes all spectra (minimum 15) within a peak Apex spectrum is the reference spectrum
Matching compares the unknown apex spectrum of the peak with a reference spectrum in a library
Dr. Shulamit Levin
Absorbance
200.00
240.00
nm
280.00
320.00
Dr. Shulamit Levin
Absorbance
230.00
250.00
270.00
290.00
310.00
nm
Dr. Shulamit Levin
Methylparaben Ethylparaben
Absorbance
200.00
240.00
nm
280.00
320.00
230.00
250.00
nm
270.00
290.00
Absorbance
1.2 nm
Analyte and one impurity spectra from 245 to 275 nm 1.2 nm resolution
246.00
254.00
262.00
270.00
nm
Dr. Shulamit Levin
500.00
400.00
nm
0.40 0.30
Zoomed Chromatogram
5/6 3
15.00
7 8 9
AU
10
4
Minutes
20.00
25.00
30.00
350.00
400.00
450.00
nm
500.00
550.00
0.02
28.982
10.00 8.00
Peak is Pure
0.01
4.00 2.00
0.00 28.50 28.60 28.70 28.80 28.90 29.00 29.10 29.20 29.30 29.40 29.50
0.00
Minutes
Degrees
6.00
0.04
0.03 AU
16.726
30.00
0.02 10.00 0.01 0.00 16.40 16.50 16.60 16.70 16.80 16.90 17.00 17.10 17.20 17.30 17.40
0.00
Minutes
Dr. Shulamit Levin
Degrees
20.00
Peak tail
}
400.00 450.00
350.00
nm
500.00
550.00
0.05 AU
Peak tail
0.00
16.00
16.50
Minutes
17.00
17.50
0.00
Th = 200 PW = 45
16.40 16.60 16.80 17.00 17.20 17.40 17.60
Minutes
0.00
Th = 10 PW = 45
16.40 16.60 16.80 17.00 17.20 17.40 17.60 Minutes
0.00
Th = 50 PW = 45
16.40 16.60 16.80 17.00 17.20 17.40 17.60 Minutes
Extracted chromatograms
200.00
220.00
240.00
nm
260.00
280.00
250.00 300.00 350.00 9.00 10.00 11.00 12.00 Minutes 13.00 14.00
Dr. Shulamit Levin
AU
0.005
0.000
11.00
Minutes
12.00
13.00
14.00
Non-Linearity Effects
Spectra collected at low portions of the peak are different than those at around the apex
SI 1 SI 2 SI 3 SI 4 SI 5 SI 6 SI 7 SI 8 SI 9 SI 10SI 11SI 12SI 13 380.00
nm 220.00
3.00 AU 2.00 1.00 0.00 7.00 7.50 8.00 8.50 9.00 Minutes 9.50 10.00 10.50
nm 220.00
0.60 0.40 AU 0.20 0.00 8.00 8.50 9.00 Dr. Shulamit Levin
Minutes