Standard operating procedure for operation and calibration of Automatic Potentiometric Titrator

 
1.0   Title:
            Standard operating procedure for operation and calibration of Automatic 
             Potentiometric Titrator
2.0       Purpose:
              To lay down the standard operating procedure for the operation and calibration of                    
             Automatic Potentiometric Titrator.
 
3.0.      Scope:
            This procedure shall be applicable for operation and calibration of Automatic
              Potentiometric Titrator determination at Fermenta Biotech Ltd.
 
4.0       Responsibility: Research Associate/ Research Scientist .
 
5.0  Procedure :           
5.1        Switch on the power supply to instrument and printer.
5.2           Display— SPECTRALAB AT38C+pH STAT
5.3        Filling of Burette:
5.3.1     Keep the Reservoir bottle containing Titrant solution on the backside of the syringe.
5.3.2     Insert the filling tube in the Titrant solution in the bottle. For time being keep the
     dosing end  of Ejection tube in an empty beaker.
5.3.3     Press <Aux Function> key and press ENTER successively till Display shows
     Rinse Burette?
     Yes = Go / No = Enter
5.3.4     Press <Go> key for Yes. Rinsing will start. The corresponding message “Rinsing Eject”
      and “Rinsing Fill” will be displayed as the rinsing operation is in progress.
5.3.5     The entire tubing, valve and syringe will be in ready position, when display shows
     “SPECTRALAB AT-38C” If air bubbles are seen, carry out rinsing again and during this                                process gently tap the tubing to drive away the air bubble.
              5.4.       Calibration of pH electrode:
5.4.1     Press <Monitor> key. Display will show “Monitor/Calibration” To perform the
      calibration,  press ENTER.
      Display—Calibration?
      Yes = Go / No = Enter
5.4.2     Press <Go> key.
5.4.3     Display—“Temp Auto/Manual    1/0” Press 1.
5.4.4     Display—“Standard Buffer pH 7.0” Clean the pH electrode and dip it in pH 7.0
      buffer  and press ENTER. When calibration with pH 7.0 buffer is completed,
      beep  appears.
      Press ENTER.
5.4.5        Display—“Standard Buffer pH 4.0” Clean the pH electrode and dip it in pH 4.0
             buffer and press ENTER. When calibration with pH 4.0 buffer is completed,
             beep appears.
      Press ENTER.
5.4.5           Display—“Standard Buffer pH 9.2” Clean the pH electrode and dip it in pH 9.2 buffer
              and  press ENTER. When the calibration with pH 9.2 buffer is completed, beep appears.
      Press ENTER.
 
5.4.7      Display—Calibration Report? Yes = Go / No = Enter
5.4.8      Press <Go> key for printed report. Clean the pH electrode with D/W.
 
5.5       Titration for Normality of NaOH:
5.5.1    Select SVD method by pressing <Method> key successively till SVD Method appears
            on the display. Press ENTER. The desired method is now selected.
5.5.2    Select Normality mode by pressing <Mode> key successively till Normality Mode
            appears on the display. Press ENTER, corresponding LED will light up.
5.5.3    Press <Sample Data> key and feed the details sush as Titration type, Titrant, Standard wt.
5.5.4    Press <Formula> key, select the Formula No. 1. and feed the details such as
            Std purity, Equivalent wt.etc
5.5.5    Press <Method & Parameter> key. And feed the details such as ML eject volume,
            Dosing speed etc.
5.5.6    Weigh the suitable quantity of Std. KHP and transfer it in the titration vessel. Dissolve it
            in  50ml of D/W. Attach the vessel to Stirrer assembly with pH electrode and
            ejection  tube dipped in the KHP solution.
5.5.7    Start the stirring to dissolve the KHP.
5.5.8    Press <Method & Go> keys to initiate the titration.
5.5.9    Progress of titration can be seen on Display.
5.5.10  After the first beep, stop the titration by pressing <Hold & Quit> keys.
5.5.11  Take the Document Report.
 
5.6       Enzyme Activity Determination:
 
5.6.1    Select pH STAT method by pressing <Method> key successively till pH STAT
            Method  appears on the display. Press ENTER. The desired method is now selected.
5.6.2    Select Sample mode by pressing <Mode> key successively till Sample Mode appears
            on  the display. Press ENTER, corresponding LED will light up.
5.6.3    Press <Sample Data> key and feed the details such as Titration type, Sample name,
            Batch  No., Operator's name etc
5.6.4    Press <Method & Parameter> keys.
            Display—ML Eject 1             00.1
                          SET/CLEAR/ENTER
            Feed the desired Ejectable volume using numerical keys.
5.6.5    Press ENTER.
            Display—ML Eject  2             00.1
                          SET/CLEAR/ENTER
            Feed the desired Ejectable volume using numerical keys.
5.6.6    Press ENTER.
            Display—Dose Speed 1           1.0
            Feed the desired time in seconds using numerical keys.
5.6.7    Press ENTER.
            Display—Dose Speed 2           1.0
            Feed the desired time in seconds using numerical keys.
5.6.8    Press ENTER.
            Display— Time ?
 
                              0 = Min / 1 = Sec.
            Select the titration time in Mins or in Sec. By pressing 0 or 1 key.
5.6.9     Press ENTER.
             Display—Titration Time            10
                             SET/CLEAR/ENTER
             Set the required titration time using numerical keys.
5.6.10   Press ENTER.
              Display—Storing Interval         1.0
                             SET/CLEAR/ENTER
             Set the storing interval time using numerical keys.
5.6.11   Press ENTER.
             Display—Initial Time          2.0
                            SET/CLEAR/ENTER
             Set the initial time which is to be used for ML EJECT 1 and DOSE SPEED 1.
             Note that the time selected and corresponding ML consumption will be deducted
             from the total titration time and Total ML consumed in Formula No. 1 & 2.
5.6.12   Press ENTER.
             Display—Solution in Burette      Base
                             Acid = 1 / Base = 0
             Select the required solution by pressing 0 or 1 key.
5.6.13   Press ENTER.
             Display—Set pH         Off pH
                             SET/CLEAR/ENTER
             Set the required pH value using numerical keys.
5.6.14   Press ENTER.
             Display—Formula No       ??
                             SET/CLEAR/ENTER
             Select the required Formula No. (1 to 3) using numerical keys.
               Press ENTER.
5.6.15   Feed the details like Blank ml if applicable.
5.6.16   Feed the enzyme sample's wt or vol. Press ENTER
5.6.17   Feed the normality of NaOH using numerical keys.
5.6.18   Take the reaction mixture in beaker and keep it in block at desired temperature.
5.6.19   After reaching the temperature, add enzyme and start the titration by pressing
            <Method  & Go> keys.
5.6.20   Progress of reaction can be seen on display.
5.6.21   After the beep, take the report of enzyme activity.
 
5.7.       Switching off the instrument:
5.7.1     Before switching off the instrument, empty the burette.
5.7.2     To empty the burette, Keep the filling tube out of titrant solution. For time being keep
             the dosing and of the Ejection tube in an empty beaker.
5.7.3     Press <Aux Function> key and press ENTER sucessively till Display shows
             Rinse Burette?
             Yes = Go / No = Enter
 
 
5.7.4      Press <Go> key for Yes. Rinsing will start. The corresponding message
    “Rinsing Eject” & “Rinsing Fill” will be displayed as the rinsing operation is in  
              progress.
5.7.5      The entire tubing, valve and syringe will become empty, when display shows
     “SPECTRALAB AT-38C”
5.7.6      Now, switch off the current supply to the instrument.
 
6.0       ABBREVIATIONS:
           

gm	:	grams
ml	:	milli liter
nm	:	nano meter
mg	:	milli gram
UV	:	Ultra Violet
cm	:	centimeter
C	:	degree Celsius
v/v	:	volume/volume
SOP	:	Standard operating procedure

 
7.0 RECORDS : Nil
 
8.0 REFERENCES : Instrument Manual