UC Berkeley College of Chemistry
Chemistry 105
Instrumental Methods in Analytical Chemistry
Reverse-Phase High-Performance
Liquid Chromatography Analysis of
Biomolecules
Short Report
Lab Partners:
Author:
Jonathan Melville
Jake Precht, Phuong Tran, and Jeremy Hsu
Graduate Student Instructors:
Richard Cooper & Daniel Mortensen
April 23, 2014
1
Theory
RP-HPLC, or Reverse-Phase High-Performance Liquid Chromatography is a type of chromatography the features a liquid mobile phase, higher resolving power than traditional
liquid chromatography, and a specific combination of stationary- and mobile-phases that
results in separations that are the opposite of a “traditional” HPLC. Chromatography, in general, utilizes the flow of a “mobile-phase” containing the analyte through
a “stationary-phase” – depending on the intermolecular forces of attraction between the
individual analytes and the stationary- and mobile-phases, the various components of
the sample will exit the chromatographic “column” at different times[?] . A substance
that experiences greater intermolecular forces of attraction to the stationary-phase will
be slowed, and will exit the column later than a substance without such attraction. If
the mobile-phase is a liquid (as opposed to, say, a gas), the process is considered “liquid
chromatography”; HPLC improves upon this process by pumping a pressurized liquid
mobile-phase through an adsorbant stationary-phase, yielding far greater separation and
resolving power than regular gravitationally-run liquid chromatography. The labelling
“reverse-phase” is far more arbitrary in origin – historically, chromatographic columns
utilized a polar stationary-phase and a nonpolar mobile-phase; “reverse-phase chromatography”, by contrast, uses a nonpolar stationary-phase and a polar mobile-phase, resulting
in an inverted separation from “normal-phase” chromatography: more polar analytes will
tend to elute earlier due to decreased interaction with the now-nonpolar stationary-phase.
Because of its versatility, ease-of-use, and superior separation, HPLC is one of the
most commonly-used classes of chromatography (itself one of the most common analytical
methods), along with other methods like gas chromatography and column chromatography.
2
Results and Discussion
This section contains only tabulated results from the Appendix. Derivations can be found
in Appendix A on page 5. Graphs can be found in Appendix B on page 12. Raw data
1
Figure 1: Methionine, a nonpolar amino acid and one of our peptides
can be found in Appendix A.1 on page 5.
2.1
Results
Due to experimental failures that are detailed further in the appendix, we were unable
to absolutely and quantitatively determine the concentration of peptides in our unknown
sample. However, we were able to determine the ratios of peptides in our sample relative
to the concentration of our calibration samples:
Peak 1:
4.13498
40.6316
+ 0.051785
0.012312
2
= 1690. ppm (relative)
Peak 2:
11.96411
15.3068
+ −0.02443
0.03288
2
2.2
= −131.3 ppm (relative) (?!)
Accuracy and Error
Of course, our accuracy in this experiment was exceedingly low. Ignoring the fact that we
are missing a crucial piece of data to turn our relative concentration values into absolute
ones (due to a failure for the instrument to read our sample, no less), the data we do have
conspires to give us abysmal fits that lead to nonsensical data. A brief examination of
our calibration curves shows that our error bars are gigantic and our R2 values abysmal
(even, in one case, producing a negative slope that directly caused the apparently negative
concentration of peptide seen above). Because of the vast amounts of missing data points
in our tables (every N/A was a point not found on the instrument), we are simply not
able to calculate anything with any degree of confidence.
2
Figure 2: Bradykinin, a blood-vessel-dilating peptide that is comparatively polar
2.3
Discussion
While we cannot make many quantitative statements about the success of our experiment,
we can still state certain facts with a reasonable degree of certainty. We can, for instance,
determine the identity of the first and second peaks eluted by looking at the structures
of the two peptides analyzed.
Methionine, the first peptide, is a rather small essential amino acid and is largely
nonpolar in character due to the presence of a fairly lengthy alkyl chain. Compared
to the much larger bradykinin, enriched with numerous amines, amides, imines, and
aromatic groups, we can say with a reasonable amount of certainty that the first peak
is that of bradykinin and the second is methionine. Because we are performing reversephase HPLC, our stationary phase is a rather waxy resin with long alkyl chains that
give it significant nonpolar character, whereas our mobile phases are comparatively polar
in nature. As a result, the more nonpolar methionine is more slowed by intermolecular
interactions with the stationary phase, allowing the more polar bradykinin to elute first.
Our data was analyzed using the HPLC’s built-in light source, an Agilent 1100/1200
Diode Array Detector that filters light from a tungsten-filament bulb through a coupling
lens, a deuterium lamp, an achromatic lens, a holmium oxide filter, a support lens, a
flow cell, and a spectro lens befor finally passing through a programmable slit and onto
a grating which projects a final spectrum[3] . Because our sample only called for UV
irradiation and not visible light (a detail noticeable in the ACQ.TXT data reports), only
the deuterium lamp is used.
Because of the data loss that plagued this entire experiment, we are unable to calculate the niceties of our isocratic separations, such as the number of theoretical plates.
3
For the exact same reasons, we are unable to determine the amount of peptide in our
unknown sample. We were able to determine the relative concentrations (1690. ppm and
-131.3 ppm) by using the relative response factor, but even this approach is unhelpful due
to abysmal correlation coefficients (ranging from 0.65 to 0.05) and simply nonsensical answers (negative concentration?!). While it is possible that our poor outputs are the result
of detection limits and exceeding the linear range of the instrument, it is unlikely that our
samples deviated so significantly from reasonable values that virtually none of them were
able to construct a decent fit. Changing the wavelength of the detector would change
the response factors of our analytes; methionine, containing little to no π-conjugation,
is likely not an exceptional absorber compared to bradykinin, which has large, extended
πsystems that likely absorb electrons of UV/Vis light readily. This also explains the
comparatively lower absorption values of methionine compared to bradykinin.
Nevertheless, the many flaws that riddled this experiment were likely a result of poor
experimental preparation and instrumental methods rather than shortcomings of the
HPLC instrument itself. Because our group was one of the last groups to complete this
lab, we were forced to make do with insufficient quantities of peptide starting material,
and whilst coordinating our sample preparation with another group we inadvertently
screwed up the preparation of several of our samples. We most likely failed to get all
of our results out of the HPLC due to a combination of this experimental error and
incorrect setup of the HPLC methods, causing several of our key analytical data points
to be dropped. Worst of all, we were unable to recreate and rerun our samples because
we had run out of lab time over the semester with which to attempt to fix our mistakes.
HPLC is a useful analytical method, even over other comparable separation methods
like electrophoresis or other forms of chromatography, because it combines the relative
ease-of-use of many chromatographic methods with the high-precision data that many
more qualitative chromatographic methods lack.
4
References
[1] SHU HPLC ; Yuri Kazakevich. http://hplc.chem.shu.edu/ (accessed April 22, 2014).
[2] Diode
Array
Detector
Optimization
[Online];
Agilent
Technologies.
https://www.chem.agilent.com/Library/slidepresentation/Public/2%20Diode%20Array%20Detecto
(accessed Apr 19, 2014).
A
Calculations
A.1
Peak Resolution Data
RA,B = 2 ∗
tret,A −tret,B
WA +WB
RA,B =peak resolution
tret =retention time
W =baseline width
A.1.1
1000 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
C
1.134
2.311
0.202
0.166
6.3967391304
2
C
N/A
N/A
N/A
N/A
N/A
3
C
N/A
N/A
N/A
N/A
N/A
A.1.2
500 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
2.191
6.44
0.182
0.185
23.1553133515
2
A
2.022
6.06
0.207
0.179
20.9222797927
3
A
2.076
6.156
0.172
0.176
23.4482758621
5
A.1.3
100 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
2.195
6.386
0.169
0.184
23.7450424929
2
A
2.063
6.165
0.18
0.19
22.172972973
3
A
2.044
6.241
0.161
0.199
23.3166666667
4
B
N/A
2.19
0.151
0.157
7.025974026
5
B
N/A
2.011
0.172
0.177
3.0601719198
6
B
N/A
2.125
0.148
0.161
4.2459546926
7
C
1.112
2.198
0.164
0.149
6.9392971246
8
C
N/A
2.198
N/A
0.16
N/A
9
C
N/A
2.341
N/A
0.187
N/A
A.1.4
10 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
1.158
2.202
0.21
0.188
5.2462311558
2
A
1.465
2.039
0.165
0.204
3.1111111111
3
A
1.457
2.062
0.189
0.212
3.0174563591
A.1.5
1 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
1.176
2.478
0.17
0.173
7.5918367347
2
A
1.518
2.129
0.193
0.201
3.1015228426
3
A
1.448
2.127
0.21
0.171
3.5643044619
A.1.6
0.1 ppm std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
1.168
2.204
0.217
0.181
5.2060301508
2
A
1.481
2.067
0.127
0.185
3.7564102564
3
A
1.452
2.1
0.184
0.186
3.5027027027
6
A.1.7
Unknown Solution
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
1
A
1.176
2.2
0.175
0.204
5.4036939314
2
A
N/A
2.076
N/A
0.172
N/A
3
A
1.59
2.072
0.167
0.183
2.7542857143
A.1.8
Bradykinin std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Bradykinin Error Log
443 41e0 53462317
Method
Method started:
67 41e0 53462319
Method
CP Macro
CP Macro
21:51:21 04/09/14
0
21:51:21 04/09/14
0
21:51:21 04/09/14
500
1 Error Method started
67 41e0 5346234b
Method
0
Data not available; Data Analysis not done
d5 1da7 5346234b
ALS
21:51:21 04/09/14
Analyzing rawdata 080-2001.D
70 41de 53462349
21:50:33 04/09/14
500
Missing Vial, no Run possible
70 41de 53462349
21:50:31 04/09/14
0
1 Missing vial (80)
67 41e0 53462349
Method
line# 20 vial# 80 inj# 1
Instrument running sample Vial 80
fb3 1da7 53462349
ALS
0
21:51:23 04/09/14
0
Method completed
21:51:23 04/09/14
7
A.1.9
Methionine std.
Trial
Method
Peak 1 (min)
Peak 2 (min)
Peak 1 width
Peak 2 width
Resolution
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Methionine Error Log
443 41e0 534625d5
Method
Method started:
67 41e0 534625d7
Method
ALS
Method
CP Macro
CP Macro
0
22:02:57 04/09/14
0
22:02:57 04/09/14
500
1 Error Method started
67 41e0 53462603
22:02:57 04/09/14
0
Data not available; Data Analysis not done
d5 1da7 53462603
A.2
22:02:56 04/09/14
Analyzing rawdata 081-2201.D
70 41de 53462601
22:02:15 04/09/14
500
Missing Vial, no Run possible
70 41de 53462601
22:02:13 04/09/14
0
1 Missing vial (81)
67 41e0 53462601
Method
line# 22 vial# 81 inj# 1
Instrument running sample Vial 81
fb3 1da7 53462600
ALS
0
22:02:59 04/09/14
0
Method completed
22:02:59 04/09/14
Bradykinin and Methionine Calibration
These calibration curves could not be created, due to a lack of any data (whatsoever)
from their respective samples1 . As a result, we are completely unable to create calibration
curves for these analytes, as well as calculating LoD or quantitative determination of the
unknown peptide mixture, due to the lack of a known standard to use as a reference.
Nevertheless, we will attempt to answer as many of these questions as possible using
1
See error logs on pages 7 and 8.
8
relative values.
Based on the error logs, it is likely that human error was responsible for this data
loss, as the samples simply failed to run. This could be caused by an incorrect method or
by physical misplacement of the samples into the HPLC chamber. Whatever the cause,
however, there is no way for us to make up for this lost information (by interpolation or
the like) because there is simply no data to interpolate from.
A.3
fi =
Response Factor Calculation
Ai
f
Astd std
Because of the limitations detailed earlier regarding the peptide standards, we are
unable to calculate absolute response factor and must settle for relative response factor:
fri =
fi
fstd
=
Ai
Astd
We can calculate this value by plotting analyte concentration against response (in the
form of area or height) and taking the slope of the best-fit line.
Unfortunately, in this instance “best-fit” may be a misnomer. “Mediocre-fit” is probably more accurate.
9
10
Based on these graphs, we receive the following relative response factors (averaging
out over multiple injections; injections were plotted separately because the data points
were clearly stratified):
Peak
fri,1
fri,2,3
fri,avg
Peak 1
.012312 .051785 .03205
Peak 2
.03288
A.4
-.02443
.00844
Relative Unknown Concentrations
Using the previously calculated relative response factors, it is possible to calculate “relative unknown concentrations” in terms of the calibration solutions we created for curvefitting. Because our “1000 ppm” standard does not actually contain 1000 ppm of both
peptides, these “relative concentrations” are ratios to the actual concentration of the
standards created, something we could determine quantitatively if our reference values
had not been lost.
Peak 1:
40.6316
4.13498
+ 0.051785
0.012312
2
= 1690. ppm (relative)
Peak 2:
11.96411
15.3068
+ −0.02443
0.03288
2
= −131.3 ppm (relative) (?!)
11
A.5
Detection Limits
Once again, without concrete values to pin our relative numbers to, we cannot quantitatively determine this value. Qualitatively, however, a cursory glance at the error bars of
our “calibration curves” assures us that hitting the detection limit is likely the least of
our worries.
B
Chromatographs
12
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\071-0201.D
Sample Name: 1000Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
2
Acq. Instrument : 305 Lat HPLC
Location : Vial 71
Injection Date : 4/9/2014 2:59:51 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
1.134
DAD1 A, Sig=254,4 Ref=360,100 (071-0201.D)
mAU
60
50
2.311
40
30
20
2.168
10
0
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.134 BB
0.1262 550.56732
65.42467 59.5121
2
2.168 BV
0.2501 100.07494
4.98669 10.8173
3
2.311 VB
0.1326 274.49249
33.14954 29.6705
Totals :
925.13476
103.56089
=====================================================================
*** End of Report ***
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\071-0202.D
Sample Name: 1000Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
2
Acq. Instrument : 305 Lat HPLC
Location : Vial 71
Injection Date : 4/9/2014 3:11:27 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
DAD1 A, Sig=254,4 Ref=360,100 (071-0202.D)
mAU
-1.5
-2
-2.5
-3
-3.5
-4
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
No peaks found
=====================================================================
*** End of Report ***
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\071-0203.D
Sample Name: 1000Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
2
Acq. Instrument : 305 Lat HPLC
Location : Vial 71
Injection Date : 4/9/2014 3:23:03 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
DAD1 A, Sig=254,4 Ref=360,100 (071-0203.D)
mAU
0
-0.25
-0.5
-0.75
-1
-1.25
-1.5
-1.75
-2
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
No peaks found
=====================================================================
*** End of Report ***
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\072-0401.D
Sample Name: 500Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
4
Acq. Instrument : 305 Lat HPLC
Location : Vial 72
Injection Date : 4/9/2014 3:45:31 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
6.440
DAD1 A, Sig=254,4 Ref=360,100 (072-0401.D)
1.209
mAU
50
6.697
40
30
0
4.843
2.113
10
2.501
2.191
20
-10
0
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.209 BB
0.1403 495.95142
53.40386 31.4287
2
2.113 BV
0.2614 103.35991
4.75820
6.5500
3
2.191 VV
0.0857
80.62334
12.10583
5.1091
4
2.501 VV
0.1670
65.41753
5.91853
4.1455
5
4.843 BB
0.1155
8.37328
1.09158
0.5306
6
6.440 BV
0.1376 533.22046
56.76577 33.7905
7
6.697 VV
0.1579 291.07367
26.97762 18.4455
Totals :
1578.01961
305 Lat HPLC 4/9/2014 3:55:43 PM Chem 105
161.02139
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Sample Name: 500Standard
=====================================================================
*** End of Report ***
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\072-0402.D
Sample Name: 500Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
4
Acq. Instrument : 305 Lat HPLC
Location : Vial 72
Injection Date : 4/9/2014 3:57:07 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.875
DAD1 A, Sig=254,4 Ref=360,100 (072-0402.D)
mAU
10
0
2.022
2.122
-10
1.513
-30
6.332
6.060
-20
-40
-50
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.875 BV
0.3477 1113.19348
39.63150 48.5831
2
1.513 VV
0.0415
15.75548
5.48666
0.6876
3
2.022 VV
0.2473 594.30695
30.24431 25.9373
4
2.122 VV
0.1684 382.46872
28.53759 16.6921
5
6.060 BV
0.2640 118.48067
6.28218
5.1709
6
6.332 VV
0.1934
67.11346
4.67787
2.9290
Totals :
2291.31877
305 Lat HPLC 4/9/2014 4:07:19 PM Chem 105
114.86010
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\072-0402.D
Sample Name: 500Standard
=====================================================================
*** End of Report ***
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Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\072-0403.D
Sample Name: 500Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
4
Acq. Instrument : 305 Lat HPLC
Location : Vial 72
Injection Date : 4/9/2014 4:08:42 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.873
DAD1 A, Sig=254,4 Ref=360,100 (072-0403.D)
mAU
10
-10
6.156
-20
6.485
1.893
1.969
2.076
2.149
0
1.450
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.873 BV
0.3515 1115.88684
39.03070 48.8070
2
1.450 VV
0.0659
5.07510
1.28902
0.2220
3
1.893 VV
0.1489 300.45795
26.36786 13.1415
4
1.969 VV
0.0733 139.63464
26.57147
6.1074
5
2.076 VV
0.0780 158.51370
28.01682
6.9331
6
2.149 VV
0.1589 333.29025
27.98861 14.5776
7
6.156 BV
0.3180 147.91566
6.25299
6.4696
8
6.485 VV
0.2101
85.54984
5.36173
3.7418
Totals :
2286.32399
305 Lat HPLC 4/9/2014 4:18:57 PM Chem 105
160.87920
Page
1 of 2
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\072-0403.D
Sample Name: 500Standard
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 4:18:57 PM Chem 105
Page
2 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\073-0601.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
6
Acq. Instrument : 305 Lat HPLC
Location : Vial 73
Injection Date : 4/9/2014 4:31:13 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
1.146
DAD1 A, Sig=254,4 Ref=360,100 (073-0601.D)
mAU
40
6.386
30
2.110
10
0
2.462
2.195
6.667
20
-10
0
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.146 BB
0.1168 353.13898
47.47415 39.8047
2
2.110 BV
0.2295
97.19534
5.16706 10.9555
3
2.195 VV
0.0838
78.58721
12.42897
8.8581
4
2.462 VV
0.1476
74.40478
7.25996
8.3867
5
6.386 BV
0.1241 202.56482
23.59933 22.8325
6
6.667 VV
0.1476
81.28827
8.06572
9.1626
Totals :
887.17940
305 Lat HPLC 4/9/2014 4:41:26 PM Chem 105
103.99518
Page
1 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\073-0601.D
Sample Name: 100Standard
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 4:41:26 PM Chem 105
Page
2 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\073-0602.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
6
Acq. Instrument : 305 Lat HPLC
Location : Vial 73
Injection Date : 4/9/2014 4:42:51 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.852
DAD1 A, Sig=254,4 Ref=360,100 (073-0602.D)
mAU
10
0
2.063
2.138
-10
6.165
-20
1.455
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.852 BV
0.3558 1071.70447
38.37623 56.6523
2
1.455 VV
0.0866
10.96265
1.81107
0.5795
3
2.063 VV
0.2603 493.35745
24.36183 26.0798
4
2.138 VV
0.1800 244.35648
22.62403 12.9171
5
6.165 BV
0.2528
71.34130
3.80328
3.7712
Totals :
1891.72234
90.97643
=====================================================================
305 Lat HPLC 4/9/2014 4:53:05 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\073-0603.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
6
Acq. Instrument : 305 Lat HPLC
Location : Vial 73
Injection Date : 4/9/2014 4:54:31 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.820
DAD1 A, Sig=254,4 Ref=360,100 (073-0603.D)
mAU
10
0
1.880
2.044
-10
6.241
-20
1.464
1.568
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.820 BV
0.3312 995.76886
37.81387 51.8885
2
1.464 VV
0.0901
27.89626
4.16753
1.4536
3
1.568 VV
0.0524
25.14947
7.63329
1.3105
4
1.880 VV
0.1588 264.83774
21.33795 13.8004
5
2.044 VV
0.2770 533.56476
23.49139 27.8035
6
6.241 BV
0.2922
71.83978
3.27133
3.7435
Totals :
1919.05686
305 Lat HPLC 4/9/2014 5:04:46 PM Chem 105
97.71537
Page
1 of 2
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\073-0603.D
Sample Name: 100Standard
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 5:04:46 PM Chem 105
Page
2 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\074-0801.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
8
Acq. Instrument : 305 Lat HPLC
Location : Vial 74
Injection Date : 4/9/2014 5:17:05 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105B.M
Last changed
: 4/2/2014 2:00:08 PM by Chem 105
DAD1 A, Sig=254,4 Ref=360,100 (074-0801.D)
1.108
mAU
40
30
2.104
10
0
2.471
2.190
20
-10
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.108 BB
0.1259 377.30368
46.85508 58.6071
2
2.104 BV
0.2398 110.27272
5.59915 17.1288
3
2.190 VV
0.0810
81.81421
13.08161 12.7083
4
2.471 VV
0.1572
74.39433
6.93048 11.5558
Totals :
643.78493
72.46632
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 5:27:18 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\074-0802.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
8
Acq. Instrument : 305 Lat HPLC
Location : Vial 74
Injection Date : 4/9/2014 5:28:44 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105B.M
Last changed
: 4/2/2014 2:00:08 PM by Chem 105
DAD1 A, Sig=254,4 Ref=360,100 (074-0802.D)
mAU
0
-10
-20
2.011
2.089
-30
1.477
-40
-50
-60
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.477 BV
0.1058
29.45708
4.00656
4.0340
2
2.011 VV
0.2417 400.07516
21.64254 54.7883
3
2.089 VV
0.1815 300.68777
20.94538 41.1777
Totals :
730.22001
46.59448
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 5:38:57 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\074-0803.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line :
8
Acq. Instrument : 305 Lat HPLC
Location : Vial 74
Injection Date : 4/9/2014 5:40:23 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105B.M
Last changed
: 4/2/2014 2:00:08 PM by Chem 105
DAD1 A, Sig=254,4 Ref=360,100 (074-0803.D)
mAU
0
-10
-20
2.005
2.125
-30
-50
0
3.097
1.469
2.506
-40
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.469 BV
0.0939
13.76783
1.96233
2.1276
2
2.005 VV
0.2091 291.84280
17.27595 45.0995
3
2.125 VV
0.1991 299.09973
19.93204 46.2209
4
2.506 VV
0.0738
8.66611
1.63504
1.3392
5
3.097 VV
0.2704
33.73269
1.53500
5.2128
Totals :
647.10917
42.34035
=====================================================================
305 Lat HPLC 4/9/2014 5:50:36 PM Chem 105
Page
1 of 1
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\075-1001.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 10
Acq. Instrument : 305 Lat HPLC
Location : Vial 75
Injection Date : 4/9/2014 6:02:55 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
1.112
DAD1 A, Sig=254,4 Ref=360,100 (075-1001.D)
mAU
30
2.198
2.112
10
0
2.451
20
-10
0
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.112 BB
0.1237 389.47934
48.47914 58.5246
2
2.112 BV
0.2201 107.68982
5.98011 16.1819
3
2.198 VV
0.0835
81.19841
12.89202 12.2012
4
2.451 VV
0.1499
87.12939
8.34221 13.0924
Totals :
665.49696
75.69348
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 6:13:09 PM Chem 105
Page
1 of 1
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\075-1002.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 10
Acq. Instrument : 305 Lat HPLC
Location : Vial 75
Injection Date : 4/9/2014 6:14:38 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
2.198
DAD1 A, Sig=254,4 Ref=360,100 (075-1002.D)
mAU
6
4
2
0
-2
-4
-6
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
2.198 BV
0.0796
78.62127
12.80807 100.0000
Totals :
78.62127
12.80807
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 6:24:52 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\075-1003.D
Sample Name: 100Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 10
Acq. Instrument : 305 Lat HPLC
Location : Vial 75
Injection Date : 4/9/2014 6:26:21 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105C.M
Last changed
: 4/2/2014 2:18:05 PM by Chem 105
2.341
DAD1 A, Sig=254,4 Ref=360,100 (075-1003.D)
mAU
6
4
2
0
-2
-4
-6
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
2.341 BV
0.0872
87.91441
13.28189 100.0000
Totals :
87.91441
13.28189
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 6:36:36 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\076-1201.D
Sample Name: 10Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 12
Acq. Instrument : 305 Lat HPLC
Location : Vial 76
Injection Date : 4/9/2014 6:48:51 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
1.158
DAD1 A, Sig=254,4 Ref=360,100 (076-1201.D)
mAU
30
2.114
2.202
10
0
2.469
20
-10
0
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.158 BB
0.1196 346.50757
45.10763 56.8083
2
2.114 BV
0.2235 105.23031
5.80513 17.2520
3
2.202 VV
0.0847
79.10258
12.35888 12.9685
4
2.469 VV
0.1539
79.11915
7.44728 12.9712
Totals :
609.95960
70.71892
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 6:59:04 PM Chem 105
Page
1 of 1
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\076-1202.D
Sample Name: 10Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 12
Acq. Instrument : 305 Lat HPLC
Location : Vial 76
Injection Date : 4/9/2014 7:00:31 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.812
DAD1 A, Sig=254,4 Ref=360,100 (076-1202.D)
mAU
10
0
2.039
2.154
-10
-20
1.465
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.812 BV
0.3228 981.17834
37.55931 62.8206
2
1.465 VV
0.0885
16.10502
2.52281
1.0311
3
2.039 VV
0.2610 365.82404
17.41731 23.4221
4
2.154 VV
0.1558 198.76657
16.83013 12.7262
Totals :
1561.87397
74.32956
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 7:10:44 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\076-1203.D
Sample Name: 10Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 12
Acq. Instrument : 305 Lat HPLC
Location : Vial 76
Injection Date : 4/9/2014 7:12:09 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.843
DAD1 A, Sig=254,4 Ref=360,100 (076-1203.D)
mAU
10
0
2.062
-10
-20
1.457
2.496
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.843 BV
0.3553 1039.12500
37.72928 66.3530
2
1.457 VV
0.0743
7.42794
1.59660
0.4743
3
2.062 VV
0.3713 513.24847
17.44443 32.7733
4
2.496 VV
0.0717
6.25335
1.26443
0.3993
Totals :
1566.05476
58.03474
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 7:22:23 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\077-1401.D
Sample Name: 1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 14
Acq. Instrument : 305 Lat HPLC
Location : Vial 77
Injection Date : 4/9/2014 7:34:36 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
1.176
DAD1 A, Sig=254,4 Ref=360,100 (077-1401.D)
mAU
30
6.265
25
20
15
2.211
5
0
2.478
10
-5
-10
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.176 BB
0.1264 351.41290
43.44933 62.2963
2
2.211 BV
0.2961
48.73400
2.07436
8.6393
3
2.478 VV
0.1485
42.87768
4.29428
7.6011
4
6.265 BB
0.1297 121.07437
14.16961 21.4633
Totals :
564.09895
63.98757
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 7:44:50 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\077-1402.D
Sample Name: 1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 14
Acq. Instrument : 305 Lat HPLC
Location : Vial 77
Injection Date : 4/9/2014 7:46:16 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.816
DAD1 A, Sig=254,4 Ref=360,100 (077-1402.D)
mAU
10
0
1.941
2.129
-10
-20
1.518
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.816 BV
0.3217 987.76434
37.45297 62.5851
2
1.518 VV
0.1092
18.65267
2.23923
1.1818
3
1.941 VV
0.2315 297.23730
17.21082 18.8331
4
2.129 VV
0.1872 274.61975
18.49891 17.4000
Totals :
1578.27407
75.40192
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 7:56:30 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\077-1403.D
Sample Name: 1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 14
Acq. Instrument : 305 Lat HPLC
Location : Vial 77
Injection Date : 4/9/2014 7:57:55 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.848
DAD1 A, Sig=254,4 Ref=360,100 (077-1403.D)
mAU
10
0
2.127
-10
-20
1.448
-30
-40
0
2
4
6
8
min
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.848 BV
0.3432 1053.58057
38.02576 64.9938
2
1.448 VV
0.0727
10.03283
2.06460
0.6189
3
2.127 VV
0.3468 557.43555
19.65986 34.3873
Totals :
1621.04895
59.75022
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 8:08:08 PM Chem 105
Page
1 of 1
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\078-1601.D
Sample Name: 0.1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 16
Acq. Instrument : 305 Lat HPLC
Location : Vial 78
Injection Date : 4/9/2014 8:20:23 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
1.168
DAD1 A, Sig=254,4 Ref=360,100 (078-1601.D)
mAU
30
2.204
6.587
2.118
2.443
10
6.306
20
0
-10
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.168 BB
0.1197 348.67422
44.36583 48.3616
2
2.118 BV
0.2066
81.84194
5.46956 11.3516
3
2.204 VV
0.0801
70.78206
11.78743
9.8176
4
2.443 VV
0.1408
81.71107
8.30485 11.3334
5
6.306 BV
0.1540
62.11704
5.75156
8.6157
6
6.587 VV
0.1565
75.84663
7.22601 10.5200
Totals :
720.97297
305 Lat HPLC 4/9/2014 8:30:37 PM Chem 105
82.90524
Page
1 of 2
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\078-1601.D
Sample Name: 0.1Standard
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 8:30:37 PM Chem 105
Page
2 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\078-1602.D
Sample Name: 0.1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 16
Acq. Instrument : 305 Lat HPLC
Location : Vial 78
Injection Date : 4/9/2014 8:32:03 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.817
DAD1 A, Sig=254,4 Ref=360,100 (078-1602.D)
mAU
10
0
2.067
-10
1.700
-20
2.498
1.481
1.583
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.817 BV
0.3190 977.10211
37.37093 59.9091
2
1.481 VV
0.0985
28.41631
4.11997
1.7423
3
1.583 VV
0.0471
16.49159
5.47909
1.0111
4
1.700 VV
0.0782
53.69276
9.76090
3.2921
5
2.067 VV
0.3344 546.24152
19.64394 33.4917
6
2.498 VB
0.0809
9.03005
1.52827
0.5537
Totals :
1630.97435
305 Lat HPLC 4/9/2014 8:42:16 PM Chem 105
77.90310
Page
1 of 2
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\078-1602.D
Sample Name: 0.1Standard
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 8:42:16 PM Chem 105
Page
2 of 2
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\078-1603.D
Sample Name: 0.1Standard
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 16
Acq. Instrument : 305 Lat HPLC
Location : Vial 78
Injection Date : 4/9/2014 8:43:41 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.832
DAD1 A, Sig=254,4 Ref=360,100 (078-1603.D)
mAU
10
0
2.100
-10
-20
2.500
1.452
1.542
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.832 BV
0.3460 1031.66931
37.85192 59.5494
2
1.452 VV
0.0862
24.26524
4.54972
1.4006
3
1.542 VV
0.0426
18.93858
6.78764
1.0932
4
2.100 VV
0.3806 644.79871
20.51986 37.2187
5
2.500 VV
0.0846
12.78787
2.05389
0.7381
Totals :
1732.45971
71.76302
=====================================================================
305 Lat HPLC 4/9/2014 8:53:55 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\079-1801.D
Sample Name: Unknown
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 18
Acq. Instrument : 305 Lat HPLC
Location : Vial 79
Injection Date : 4/9/2014 9:06:09 PM
Inj :
1
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
1.176
DAD1 A, Sig=254,4 Ref=360,100 (079-1801.D)
mAU
30
25
20
6.204
15
2.115
5
0
2.495
2.200
10
-5
-10
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
1.176 BB
0.1154 297.52802
40.63152 45.2417
2
2.115 BV
0.1941
88.30638
5.59623 13.4277
3
2.200 VV
0.0826
76.47616
11.96411 11.6289
4
2.495 VV
0.1685
72.60163
6.20496 11.0397
5
6.204 BB
0.2100 122.72923
7.86909 18.6620
Totals :
657.64141
72.26591
=====================================================================
305 Lat HPLC 4/9/2014 9:16:21 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\079-1802.D
Sample Name: Unknown
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 18
Acq. Instrument : 305 Lat HPLC
Location : Vial 79
Injection Date : 4/9/2014 9:17:44 PM
Inj :
2
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.862
DAD1 A, Sig=254,4 Ref=360,100 (079-1802.D)
mAU
10
0
1.965
2.076
2.166
-10
-20
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.862 BV
0.3561 1105.59680
38.37947 64.4417
2
1.965 VV
0.2169 285.18832
16.39925 16.6227
3
2.076 VV
0.1018 133.01527
18.09304
7.7530
4
2.166 VV
0.1484 191.85429
16.65150 11.1826
Totals :
1715.65469
89.52326
=====================================================================
*** End of Report ***
305 Lat HPLC 4/9/2014 9:27:58 PM Chem 105
Page
1 of 1
min
Data File C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\079-1803.D
Sample Name: Unknown
=====================================================================
Acq. Operator
: Chem 105
Seq. Line : 18
Acq. Instrument : 305 Lat HPLC
Location : Vial 79
Injection Date : 4/9/2014 9:29:22 PM
Inj :
3
Inj Volume : 20.0 µl
Sequence File
: C:\Chem32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105.S
Method
: C:\CHEM32\1\DATA\9 APRIL 2014\CHEM105 2014-04-09 14-47-10\CHEM105A.M
Last changed
: 3/18/2014 3:50:33 PM by Chem 105
0.842
DAD1 A, Sig=254,4 Ref=360,100 (079-1803.D)
mAU
10
0
1.919
2.072
2.158
-10
1.590
-20
-30
-40
0
1
2
3
4
5
6
7
8
9
=====================================================================
Area Percent Report
=====================================================================
Sorted By
:
Signal
Multiplier:
:
1.0000
Dilution:
:
1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=254,4 Ref=360,100
Peak RetTime Type Width
Area
Height
Area
#
[min]
[min]
[mAU*s]
[mAU]
%
----|-------|----|-------|----------|----------|--------|
1
0.842 BV
0.3098 918.49908
36.23124 62.7453
2
1.590 VV
0.0526
14.37509
4.13498
0.9820
3
1.919 VV
0.1996 242.28671
16.66873 16.5513
4
2.072 VV
0.1038 112.53143
15.30680
7.6873
5
2.158 VV
0.1492 176.16068
14.98468 12.0340
Totals :
1463.85300
87.32643
=====================================================================
305 Lat HPLC 4/9/2014 9:39:37 PM Chem 105
Page
1 of 1
min