Hitachi F7000 Instruction Manual

							4.2.2 
4 - 5 
Upon selection of the Quantitation tab, the window shown in Fig. 4-4 
appears.  
For the settable range of each parameter, refer to Appendix G.   
 
 
 
Fig. 4-4  Quantitation Tab 
 
 
1 Quantitation type   
Quantitation type is selectable among Wavelength, Peak area, 
Peak height, Derivative and Ratio.    Spectra measured by the Peak 
area, Peak height and Derivative methods can be checked, 
referring to 7.19.   
(a) Wavelength  
Upon selecting ‘None’ for Calibration type in this quantitation 
method, photometric values can be obtained at the specified 
number of wavelengths for Number of wavelengths.     
A calibration curve will not be prepared.   
 
Upon selecting 1st order, 2nd order, 3rd order or Segmented 
in this quantitation method, calibration curve data and sample 
data will be obtained by one, two or three-wavelength 
calculation depending on your entry for Number of 
wavelengths.  
 
4.2.2 Quantitation 
Tab 
 
1 
2 
5 
3 
4 
7
6
8  
						

							4.2    Creating an Analysis Method 
4 - 6 
(i)  One wavelength calculation   
This is the most generally used method.    When you 
specify one wavelength and enter it at the Instrument 
tab, a calibration curve and concentration are 
determined from the data I
1 at the specified wavelength 
as shown in Fig. 4-5 (a).   
 
 
 
Fig. 4-5 (a) 
 
 
(ii)  Two wavelength calculation   
When you specify two wavelengths and enter them at 
the Instrument tab, a calibration curve and 
concentration are calculated through determination of I 
by the equation below assuming I
1 and I2 are the 
photometric values at two different wavelengths as 
shown in Fig. 4-5 (b).     
 
 
 
Fig. 4-5 (b)   
 
 
I1
Data 
I 
WL1 
Data 
I1  I
2
I 
WL1  WL2 
						

							4.2.2 
4 - 7 
(iii)  Three wavelength calculation   
As shown in Fig. 4-5 (c), assuming I
1, I2 and I3 are the 
data values at three different wavelengths, a calibration 
curve and concentration are calculated through 
determination of I by the equation below.     
Photometry is made in the order of WL1, WL2 and WL3.     
 
I = I
2 -   
 
But the following relation must hold:   
WL1 < WL2 < WL3 or WL1 > WL2 > WL3   
 
 
 
Fig. 4-5 (c) 
 
 
(b) Peak area  
Enter the peak wavelength and its accuracy for Peak apex.   
Integration method (area detection method) is selectable 
among Rectangular, Trapezoid and Romberg.    (Refer to 
“integration method” in Appendix D.2.)    Then, set a 
threshold value and sensitivity for defining the peak, referring 
to 7.6.   I
1 (WL3 - WL2) + I3 (WL2 - WL1) 
WL3 - WL1 
Data 
I1  I
2
WL1  WL2 I
3
WL3I 
						

							4.2    Creating an Analysis Method 
4 - 8 
Measurement will be carried out within the wavelength range 
set at the Instrument tab.    According to the specified 
threshold level and sensitivity, peak wavelength is sought.   
The area of the peak found within a range of the entered 
Peak apex ± wavelength accuracy is calculated, thereby 
preparing a calibration curve.   
 
 
 
Fig. 4-6    Quantitation Tab (peak area) 
 
  
						

							4.2.2 
4 - 9 
(c) Peak height  
Enter the peak wavelength and its accuracy for Peak apex.   
Measurement will be carried out within the wavelength range 
set at the Instrument tab.    According to the threshold level 
and sensitivity specified for the “Peak area” quantitation 
method, peak wavelength is sought.    A calibration curve is 
prepared using the height of the peak found within a range of 
the entered Peak apex ± wavelength accuracy.     
 
 
 
Fig. 4-7    Quantitation Tab (peak height) 
 
  
						

							4.2    Creating an Analysis Method 
4 - 10 
(d) Derivative  
Enter peak wavelength (or wavelength to be used for 
calibration curve preparation) for Peak apex.     
Measurement will be carried out within the wavelength range 
set at the Instrument tab.    A calibration curve is created 
using the derivative value at the peak wavelength.     
Assign your values for derivative parameters; Derivative 
order, Smoothing order and Number of points.   
 
 
 
Fig. 4-8    Quantitation Tab (derivative) 
 
  
						

							4.2.2 
4 - 11 
(e) Ratio  
Upon selecting Ratio, an input column for three factors (K-
factor 0, K-factor 1, K-factor 2) and a setting column for 
calculation method (+, -, ∗) are displayed.     
 
NOTE:  By this quantitation method, a calibration curve cannot 
be created.   
 
 
 
Fig. 4-9    Quantitation Tab (peak ratio) 
 
 
Calculation is made by the following equation.   
 
RATIO =    #K0 
 
# (calculation method):    +, -, ∗  
WL1, 2, 3  :  Specified wavelengths   
WL3  :  Background correcting wavelength   
A ( )  :  Measured value at specified wavelength   
 
2 Calibration type   
Set a calibration type.    For selecting factor input (when the formula 
of standard calibration curve and its factors are already known), put 
a mark in the check box for Manual calibration and enter values for 
A0, A1, A2 and A3.     
 K1 (A (WL1) - A (WL3))
K2 (A (WL2) - A (WL3)) 
						

							4.2    Creating an Analysis Method 
4 - 12 
(a) None  
Mode in which calibration curve is not used.    When this 
mode is selected with Wavelength set for Quantitation type, 
multi-wavelength measurement at up to six wavelengths is 
possible.    However, no spectrum can be displayed.   
 
(b) 1st order  
This specifies the most generally used calibration curve.     
A linear calibration curve is to be created.    The formula of 
calibration curve is as follows.   
x = A
1y + A0  
Here, x is sample concentration and y is sample data.   
 
 
 
Fig. 4-10  1st Order 
 
 
(c) 2nd order  
A calibration curve is created using a quadratic curve.     
The formula of calibration curve is as follows.   
x = A
2y2 + A1y + A0 
Here, x is sample concentration and y is sample data.   
 
 
 
Fig. 4-11  2nd Order 
Data 
K0Conc 
Data 
K0Conc  
						

							4.2.2 
4 - 13 
(d) 3rd order  
A calibration curve is created using a cubic curve.     
The formula of calibration curve is as follows.   
x = A
3y3 + A2y2 + A1y + A0 
Here, x is sample concentration and y is sample data.   
 
 
 
Fig. 4-12  3rd Order  
 
 
(e) Segmented  
A calibration curve created by connecting standard samples 
linearly according to the measured values of respective 
samples or input data.   
 
 
 
Fig. 4-13  Segmented 
 
 
Data 
K0Conc 
Data 
Conc
For the part which 
exceeds the measuring 
range, extend the line 
just before it. 
Max. No. of standards:    20  
						

							4.2    Creating an Analysis Method 
4 - 14 
NOTE:  A proper calibration curve can be created when the 
photometric value increases or decreases 
monotonously versus the concentration value.    With a 
monotonic increase, the calibration curve is created with 
standard 1 (STD1) as origin.    And with a monotonic 
decrease, the calibration curve is created with STD1 
plotted on the photometric value axis and STD5 on the 
concentration axis.    In these examples, the number of 
standards is set at 5.   
 
 
 
 
3 Number of wavelengths   
Enter the desired number of wavelengths to be used for 
measurement.  
4 Concentration unit   
Enter a concentration (in mg/mL for example) from the keyboard.   
Calibration curve with monotone increase
Calibration curve with monotone decrease
Data  Data 
Conc
Conc