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Hitachi F 2500 Manual

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Page 111

4 - 6 
 
 
 
 
 
 
 
 
 
 
 
 
Fig. 4-5  Calculation Method 
 
 
2) Peak area  
Input the peak wavelength and its accuracy.   
Measurement is made in the wavelength range 
specified at the Instrument tab.     
Peak wavelength is obtained and a calibration 
curve is prepared using the peak area of relevant 
peak.    Select an integration method among 
Rectangular, Trapezoid and Romberg.    Also set 
a threshold value and sensitivity for defining the 
peak.  
 
 
 
Fig. 4-6    Quantitation Tab (peak area)...

Page 112

4 - 7 
3) Peak height  
Input the peak wavelength and its accuracy.   
Measurement is made in the wavelength range 
specified at the Instrument tab.    Peak is 
determined within the accuracy set here and 
calculation is made in the height of that peak.     
 
 
Fig. 4-7    Quantitation Tab (peak height) 
 
 
4) Derivative  
Input a peak wavelength (or wavelength to be 
used for calibration curve preparation).   
Measurement is made in the wavelength range 
specified at the Instrument tab and a...

Page 113

4 - 8 
5) Ratio  
Upon selecting Ratio, an input column for three 
factors (K0, K1, K2) 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 =   
 
 
# (calculation method):    +, -, *   
WL1, 2, 3 :  Specified wavelengths   
WL3  :  Background correcting wavelength   
A( )  :  Measured value...

Page 114

4 - 9 
1) None  
Mode in which calibration curve is not used.   
When this mode is selected with Wavelength set 
for Quantification type, multi-wavelength 
measurement at up to six wavelengths is possible.   
However, no spectrum can be displayed.   
2) 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....

Page 115

4 - 10 
3) 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 
Ko Conc  4.2    Creating an Analysis Method  

Page 116

4 - 11 
4) 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 
 
 
 
 
Data 
Ko Conc 4.2  

Page 117

4 - 12 
 
5) 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 
 
 
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...

Page 118

4 - 13 
 
 
 
Calibration curve with monotonous increase 
 
 
 
 
 
Calibration curve with monotonous decrease 
 
 
(c)  Number of wavelengths   
When “Calibration type” is “None,” up to six 
wavelengths are selectable, and with other calibration 
types, 1 to 3 wavelengths are selectable.   
 
(d) Concentration unit  
Input a concentration (in mg/mL for example) from the 
keyboard.  
 
(e) Manual calibration  
Put a check mark in this box for creating a calibration 
curve using factors.    The number of...

Page 119

4 - 14 
(f)  Force curve through zero   
By putting a check mark in this box, a calibration curve 
is created so that its factor A
0 passes through “0” 
automatically.   
 
(g)  Digit after decimal point   
Set the number of digits after decimal point for the 
calculated concentration of the measured sample.   
The concentration value is rounded off to the set 
number of digits.     
Input range:    0 to 3   
 
(h)  Lower concentration limit/Upper concentration limit   
This is used to set a normal value...

Page 120

4 - 15 
(a) Data mode  
Select one of the following data modes.   
The Luminescence mode is for measuring 
luminescence in the biological/chemical field and only 
the sample side signal is acquired without performing 
ratio photometry.   
• Fluorescence  
• Luminescence  
• Phosphorescence  
 
(b) Wavelength mode  
Select a wavelength for creating a calibration curve.   
•  EX WL Fixed   
•  EM WL Fixed   
•  Both WL Fixed   
 
(c)  Wavelength (1 to 6)   
Input the number of wavelengths specified at the...
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