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The following data ware obtained for caffeine standards measured by HPLC:
25
15.4
Concentratio
50
75
n (mg/l)
Peak34.7
Height
53.1
(mm)
100
68
125
82.2
150
95.7
a) Make a linear plot of concentration against peak height. Include the equation
of the line and the product moment correlation coefficient.
50
0
By using both Minitab and Excel, we make a linear plot of concentration against peak
height.
The equation of the line of both method is:
y=0.6387 x +2.2933
Where,
is concentration (mg/l).
0.9942
99.4
Which is the
1 thats
mean the calibration curve in both graphs are very close to be linear.
x values.
residuals against
Concentration
25
50
75
100
125
150
Peak Height
15.4
34.7
53.1
68.0
82.2
95.7
Residuals
-2.86190
0.46952
2.90095
1.83238
0.06381
-2.40476
Table 1: The
95
y
y
values
x values, where,
values.
x is
is peak height.
confidence intervals for the line on the graph.
95
95
graph.
84.2 mm
83.6 mm
83.9 mm
84.4 mm
y=0.6387 x +2.2933
84.2=0.6387 x +2.2933
x=128.2 mg /l
2)
83.6=0.6387 x+2.2933
x=127.3 mg/l
3)
83.9=0.6387 x +2.2933
x=127.8 mg /l
4)
84.4=0.6387 x+ 2.2933
x=128.6 mg /l
value
84.2
Readings
83.6
(mm)
83.9
128.2
Concentratio
127.3
n of unknown
(mg/l)
127.8
84.4
128.6
84.0
128.0
Mean
An unknown concentration that have been given with readings in table (2).
e) Calculate the approximate random error of the mean concentration from the
equation:
y 0 2
x i x 2
b2
i
1 1
+ +
m n
Sy
S x = x
b
0
Where,
Sx
Experimental value of
concentration),
Sy
x
x0 ,
is
y0
is
is Number of replicates of
yi
is
Table (3) will show the data what we are using in the equation to
calculate the approximate random error of the mean concentration.
7
Sum
xi
yi
25
15.4
50
34.7
75
53.1
100
68.0
125
82.2
150
95.7
525
349.1
x
( ix )
(25-87.5)
= - 62.5
(50-87.5)
= - 37.5
(75-87.5)
= - 12.5
(100-87.5)
= 12.5
(125-87.5)
= 37.5
(150-87.5)
= 62.5
10937.5
y
y
( i )
3906.25
(15.4-58.18)
= - 42.8
- 1831
18.2
7.84
1406.25
(34.7-58.18)
= -23.5
- 552.3
34.2
0.25
156.25
(53.1-58.18)
= -5.1
- 26
50.2
8.41
156.25
(68-58.18) =
9.8
96
66.2
3.24
1406.25
(82.2-58.18)
= 24
576
82.1
0.01
3906.25
(95.7-58.18)
= 37.5
1406.3
98.1
10937.5
-1
- 331.8
349
26
Mean ( x )=87.5
Mean ( y )=58.18
^y
y 0 2
x i x 2
2
b
i
1 1
+ +
m n
Sy
S x = x
b
0
Where,
n=6
m=4
b=0.6381
=58.18
x =87.5
y 0=84.0208
2
xi x =10937.5
y i ^y 2
S y =
x
So,
84.0258.18 2
1 1
+ +
4 6
2.55
Sx =
0.64
0
f)
Sx 3
0
Can you find a better way of fitting a line through the date points? Make a
graph of the best fit you can find (include 95% confidence intervals). Include
the equation of the line and the adjusted R square value.
10
By using Minitab, we got the best fit with Quadratic that showed in figure (5), Which
gives as the best value of
R2
R =100
x=
Where,
b b24 ac
2a
a=0.001309
b=0.8677
c=5.340
x=
x=
x=
x=
12
84.2
Unknown
Readings
83.6
(mm)
128
Unknown
Concentratio
126.8
n
84.4
128.3
83.9
127.4
128.4
128
Unknown
Concentration from
the best
fitted plot
126.8
equation
Unknown
concentration from
linear 127.5
plot equation
128
127.4
128.8
128.3
13
N
4
4
4
Mean
128.175
127.625
0.5500
StDev
0.556
0.665
0.1291
SE Mean
0.278
0.333
0.0645
P-Value = 0.003
By using Paired T-Test, we could see that the p-Value = 0.003 which is
0.05 that
will accept the null hypothesis and there are a significant difference between the
mean concentrations that calculated from the two calibrations.
water, FW = fish in fresh water, S = silver eels, y = yellow eels. The yellow form of the
eel is usually found in fresh water. They change into the silver form before migrating
to sea water).
SWs
400
405
375
355
343
357
349
394
337
367
FWs
335
331
328
336
334
329
363
336
339
341
SWy
375
386
385
372
372
402
387
416
407
394
FWy
338
337
339
334
333
351
343
343
333
339
By using Minitab, We could make a box-and-whisker plot of the data which is shown
in figure (6).
15
By made box-and-whisker plot of SWs, FWs, SWy, and FWy, we obtained data in
table (7).
Water
Median
Interquartile Range
362
335
386
338
b) Make a column chart of the data. Include 95% confidence intervals. Indicate
which, if any, groups differ significantly (ANOVA followed by a post hoc test) from
which other groups on the plot.
By using Minitab, We could make a column chart of the data given, with 95%
confidence intervals.
With using ANOVA, We could see groups differ significant clearly.
16
DF
3
36
39
S = 15.43
Level
SWs
FWs
SWy
FWy
N
10
10
10
10
SS
18952
8572
27524
MS
6317
238
F
26.53
R-Sq = 68.86%
Mean
368.20
337.20
389.60
339.00
P
0.000
R-Sq(adj) = 66.26%
StDev
24.39
9.95
15.08
5.56
Mean Grouping
10 389.60 A
10 368.20
B
10 339.00
C
10 337.20
C
--------+---------+---------+---------+(---*----)
(---*----)
(----*---)
--------+---------+---------+---------+-40
0
40
80
--------+---------+---------+---------+(----*----)
(---*----)
--------+---------+---------+---------+-40
0
40
80
--------+---------+---------+---------+(---*----)
--------+---------+---------+---------+-40
0
40
80
17
By using Minitab, with a column chart with 95% confidence intervals, we obtained the
graph which is shown in figure (7).
By using ANOVA, We obtained the data which is show in table (8), and that contains:
i.
ii.
iii.
iv.
There is a significant different in SWs which is from SWy, FWs, and FWy.
In FWs there is a significant different from SWy.
Both FWs and FWy are same, that no significant different.
Finally, both SWy, and FWy have a significant different.
Reference:
o
18