SOP FOR OPERATING PROCEDURE OF ROTARY VACCUM EVAPORATOR

1.0              Objective:

            The purpose of this SOP is to provide written procedure for operation of Rotary Vacuum evaporator.

2.0              Scope:

            This SOP covers operating procedure of Rotary vacuum evaporator 

3.0              Responsibility:

            Junior Research officer, Research Officer: Responsible for operation of the apparatus as   per procedure.

            QA Officer/QA Manager: Review the records and governing the document.

4.0       Procedure:

4.1       Connect the assembly as given in manual.

4.2       Fill water in water bath and start heating so that temp of water should be 40 ± 2°C

4.3       Take extract in round bottom flask.

4.4       Connect to assembly so that part of the flask is dip in water bath.

4.5       Start vacuum pump set the vacuum at 470 mm of Hg.

4.6       Set the revolutions at 100 rpm.

4.7       Dry the sample till evaporates to dryness and collects the residue with suitable solvent in  vial.   

SOP FOR OPERATING PROCEDURE OF HIGH PERFORMANCE LIQUID CHROMATORGAPHY SYSTEM SHIMADZU (Model: LC 2010CHT).

1.0                       Objective:

               To provide written procedure for operation of High Performance Liquid                         Chromatography system EQU-INS-201.

2.0                       Scope:

               This SOP covers operation of High Performance Liquid Chromatography system make: Shimadzu (Model: LC 2010CHT).

3.0                       Responsibility:

Jr. Research Officer, Research Officer: Responsible for operation, Calibration and maintenance of the instrument as per procedure.

Head of Department: Responsible for calibration and maintenance, timely as per schedule.

QA Officer/ QA Manager: Review the record and governing the document.

4.0          Procedure:

4.1          Ensure that the instrument is visibly clean and free from dust.

4.2          Starting UP and Connecting Instruments and Class –VP Software:

4.2.1              Turn on the instruments.

4.2.2          After confirming that the instrument is started up, turn the PC power ON, and select and click on Start icon, Select programs and chromatography and click on ‘CLASS VP’. OR double click on ‘CLASS VP’ icon on desktop. Display shows Class VP dialog box.

4.2.3               Select and double click on the instrument icon in the Class VP dialog box.  

4.2.4              Enter a user name ‘System’ and password ‘2001’; to log in to ‘CLASS-VP’, the instrument window opens.

4.3          Click on pump icon or and fed the require flow rate and concentration of solvent.

4.4          Purging Mobile phase and Rinse Solution:

4.4.1       Click on the ‘PURGE’ icon on the control toolbar. Select the flow lines to be purged and set purge time for each.

4.4.2              Clicking on the purge button display a window showing the progress of purge, and the auto purge starts.

4.4.3              If stop the purging click ‘Mobile Phase Stop’ button.

4.4                    Click on ‘PUMP’ icon into instrument window; Select the mode of pump as Isocratic flow or low pressure Gradient, feed the flow rate and concentration of solvent in %, adds maximum pressure and minimum pressure value.

4.5                    Create a New Method File or Modify Method:

4.6.1          Choose Open or ‘New’ option from the method in File menu in the instrument window, Method dialog box display on the screen.

4.6.2        Select the commands in the Method menu from the Option tab, Select and click on Properties, Method properties dialog box display, feed the require parameters, then click on ‘OK’ button.

4.6.3         Select and click on Integration Events in method menu, Integration Events dialog box display, feed the require parameters, and then click on ‘OK’ button.

4.6.4         Select and click on Peaks/Groups in method menu, Peaks/Groups dialog box display; add Component name, Retention time etc. and then click on ‘OK’.

4.6.5       Select and click on Advanced, Advanced method option dialog box display, Select Component name, Retention time, Area, Asymmetry, and Resolution etc. 

4.6.6                 Select and click on Instrument Setup in method menu, Instrument dialog box display, and click on ‘PUMP’ button.

4.6.7              Select the mode of pump as Isocratic flow or low pressure Gradient, feed the flow rate and concentration of solvent in %, adds maximum pressure and minimum pressure value.

4.6.8               Click on ‘Oven’ button, add temperature maximum up to 60° and add Oven temperature.

4.6.9             Click on ‘Detector’ button, Select D2 lamp, Polarity, Cell temperature low, add wavelength on Channel 1 and require for Channel 2, add sensitivity and select Acquisition On, add require Run time.

4.6.10             Click on ‘Controller’ and select Degasser.

4.6.11    Click on ‘Time Program’ and select module (Pump, Oven, Detector etc.) add require time and program.

4.6.12          Select and click on System Suitability in method menu, System Suitability dialog box display, select the require parameters i.e. Area, Retention time, Asymmetry, Resolution etc. and then click on ‘OK’.

4.6.13       After completion all the parameters save method from file menu. Enter or Select path and full file name.

4.7          Create a New Sequence File or Modify Sequence :   

4.7.1               Choose Open or ‘New’ option from the Data in File menu in the instrument window, Data dialog box display on the screen, create a new folder and give name of the folder.

4.7.2           Choose Open or ‘New’ option from the sequence in File menu in the instrument window, sequence dialog box display on the screen.

4.7.3           In Run information add Sample ID name, Select method, Select Data path, and give Data file name.

4.7.4                 Add Amount values as per requirement.

4.7.5                 In Auto sampler feed the parameters, start vial, end vial, injection volume and Repetitions per run, and then click on ‘OK’, sequence file display on the screen.  

4.7.6     In sequence file feed parameter sample identification, vial number, injection volume respectively.

4.7.7              After completion all the parameters save sequence from file menu. Enter or Select path and full file name.

4.7.8            After completion all parameters, right click of the mouse, select and click on start sequence.

4.8          Creating Report Templates:

4.8.1        Click on the Edit Custom Report button or Select and click on Custom Report, Custom Report dialog box display.

 4.8.2       Right click of the mouse, select report header and feed parameters, select graph and drag                     the      graph on custom report dialog box display, peak report. After completion feed all                       parameters save template file.

4.8.3             Select print in file menu, and give print command for printing.

4.9                    Opening of Instrument Off Line:

4.9.1       Select and double click on the instrument off line icon in the Class VP dialog box.  

4.9.2               Enter a user name ‘System’ and password ‘2001’; to log in to ‘CLASS-VP’, the instrument off line window opens.

4.10        COLUMN FLUSHING:

4.10.1     After completion of analysis clean the column by the following solvents as mobile phase.

4.10.2        For reverse phase columns:

4.10.2.1          Flush with the same mobile phase, which was need for analysis for 15 minutes with  1.0 ml flow rate.

4.10.2.2          Flush with water for 30 minutes.

4.10.2.3     Then flush with methanol for 15 minutes at a flow rate of 1.0 ml / minute.

4.10.3        For normal phase columns:

4.10.3.1     Flush with the same mobile phase which was need for the analysis for 15 minutes at a  flow rate of 1.0 ml / minute.

4.10.3.2     Then flush with n -Hexane at a flow rate of 1.0 ml / minute.

4.10.4        Change over from Reverse phase to Normal phase :

4.10.4.1     After the analysis is over flush the column with water for 30 minutes, followed by             Methanol for 15 minutes.

4.10.4.2          Remove the reverse phase column (C18 or C8) and attach a dead volume instead of

4.10.4.3                              column.

4.10.4.3     Now flush the system with acetonitrile followed by chloroform and n -hexane (all 15  minutes each).

4.10.4.4     Remove dead volume and fix a normal phase column (e.g. silica)         

4.10.4.5     Continue flushing for 15 minutes.

4.10.4.6     Maintain the reverse phase column in methanol.

4.10.5        Change over from normal to reverse phase:

4.10.5.1     After the analysis is over flush the columns with n- hexane for 15 minutes.

4.10.5.2     Remove the normal phase column (C18 or C8) and attach a dead volume instead of          column.

4.10.5.3     Flush the system with chloroform followed by acetonitrile and methanol for 15            minutes.

4.10.5.4     Now fix the reverse phase column and flush the column for 15 minutes.

4.10.5.5          Continue the analysis in reverse phase. Store the normal phase column in n -hexane.

4.11           Enter the details in HPLC usage logbook as per given in Annexure –1.

5.0             Safety precaution:

5.1             After analysis run the system wash with water for 30 minutes if buffer solution used in        mobile phase followed by methanol for 15 minutes.

6.0             Routine maintenance:

6.1             Clean the instrument by dry cloth.

6.2             Clean the system with hot water without connecting the column. Than methanol and

                  finally with water.

6.3              Sonicate the suction filter with 2 M nitric acid and than water and than water.

SOP FOR OPERATING PROCEDURE OF GAS CHROMATOGRAPHY AGILENT 6890N

1.0                    Objective:

               The objective of this Operating procedure is to lay down the procedure for Operating the Gas Chromatography Agilent 6890N.

2.0                      Scope:

               This procedure covers operation of Gas Chromatography.

3.0                    Responsibility:

              Jr. Research Officer, Research Officer: Responsible for operation and maintenance of

               the instrument as per procedure.

Head of Department: Responsible for maintenance, timely as per   schedule.

QA Officer/QA Manager: Review the records and governing the document

4.0          Procedure:

4.1       

   I.     Ensure that the instrument is visibly clean and free from dust.

  II.     The room temperature should be maintained between 10 to 30°C and relative humidity 

            Should be maintained between 30 to 80 %

III.    Avoid direct exposure to air-blow from air conditioners.

IV.    Avoid exposure to direct sunlight and vibrations.

V.     Avoid from equipment which generates strong magnetic field.

4.2          Start UP Procedure:

 4.2.1       Affix the require column (Capillary or packed) in to the column oven.

 4.2.2       Open the valve of gas cylinder of Z - air, Hydrogen and carrier gas (Nitrogen or Helium) and checked the pressure and leakage with 50 % v/v Isopropyl Alcohol solution.      

 4.2.3       Now Switch ON the Mains and Instrument. Switch on the computer.

4.3          Setting Instrument Parameters:

4.3.1      After Switch On instrument will display Oven Off message. Press oven key and press on key. The instrument started temperature increase of previous set temperature value.

4.3.2     Press Oven key, Add initial temperature, initial time and Rate (if required) with numerical key and press Enter key to confirm the vale.

4.3.3     If adding rate 1 the next oven program display, feed the required temperature, time and rate and then press Enter key to confirm the value. If column (Oven) temperature is   Isothermal condition the rate is off.  

4.3.4      Set the Oven Final Temperature below 20° C of Maximum Column temperature or as per procedure requirement.

4.3.5      Press ‘Back inlet’ key select mode split or split-less, add require inlet temperature with numerical key and press ‘Enter’ key to confirm the value.

4.3.6         Press ‘Front Det.’ key, add require detector temperature with numerical key and press ‘Enter’ key to confirm the value. Now increase the temperature, if temperature comes out at 150° C message display ‘FLAME OUT’.

4.3.7             Press ‘Front Det.’ Key and select Flame with ▲ and ▼ key select flame on with press ‘On’ key. Igniting message display and ignite the flame with gas lighter on detector side. The display shows the flame indicator in a numerical value.

4.3.8      Press ‘Status’ key and check the fed parameters again with ▲ and ▼ key to confirm the value. Now instrument is ready, the message will display “Instrument ready for Injection’.

4.3.9           Press “Signal 1” key select Zero with▲ and ▼ key, press ‘On’ key to flame numerical value should be zero and base line detection started to zero mv.

4.3.10  Now inject the solution with syringe. If used manual operation press ‘START’ key immediately after inject the solution.

4.4                     Software Operation:

4.4.1    Double click on the instrument1 (online) & instrument1 (offline) icon from the desktop;

             create new method from instrument1 (offline) and load method in instrument1 (online)

 from file menu. Apply the chromatographic condition and enter ok. And view the run

 signal window, After completion of the run, and open the instrument offline select the file 

 and] integrate the peak using integration menu, go to calibration task icon and enter the

             component name and click on ok, then print the chromatogram by clicking print option

             from file menu.

4.4.2  Condition the column at 280°C for 10 min. Enter the programming condition of the                        sample through micro syringe, when G.C shows ready signal.

4.4.3  Cool the G.C after completion of the analysis & switch it off. Shut down the computer, 

             mains power supply. Close the gas supply from the distribution panel & from cylinder.

5.0       Routine Maintenance:

5.1       Clean the all out side part of instrument.

5.2       Clean the injector and injector liner. Change the glass wool of injector liner. Change the

 septa of injector port if  required.

5.3         Check the all connection of gases for leakage.

SOP FOR OPERATION AND CALIBRATION OF TABLET DISSOLUTION APPRATUS

1.0       Objective

The objective of this Standard Operating Procedure is to lay down the procedure for operating the Tablet Dissolution Apparatus Model No TDT-08L.

2.0       Scope

This SOP covers operating procedure of Tablet Dissolution Apparatus Model No TDT-

08L

3.0       Responsibility

Jr. Research Officer, Research Officer: Responsible for operation, calibration and maintenance of the instrument as per procedure.

Head of Department: Responsible for calibration and maintenance, timely as per schedule.

QA Officer/QA Manager: Review the records and governing the document.

4.0       Procedure

4.1       Ensure the instrument is Clean & Free from Dust

4.2       System Start Up

4.2.1    Switch ON The ETC-11L instrument by pressing the power button

4.2.2    Now, Switch ON the Power switch provided on rear side of TDST-08L the instrument will initialize by displaying a power flash screen, which will flash once.

4.2.3    After the power flash screen will be displayed showing the last protocol no selected, RPM, Bath temperature, and temp by external probe.

4.2.4    Raise the instrument by pressing the lift UP Key provided in the front panel TDT-08L.

4.2.5    Remove the bath Top Plate and fill the bath with R.O or distilled water till the bottom of the inlet Nozzle. Once the tube of the inlet nozzle is filled with water, ensure the priming of the pump. Then fill water till the level marked. Place the bath top plate and tighten the

plate retaining Knob

4.2.6    Place the test vessels filled with the media.

4.2.7    If required check the centering of the Test Vessel.

4.2.8    Set the test parameter as desired.

4.2.9    Parameter Setting:

RPM Setting:

In this parameter rotation per minute of the shaft are set from 25 to 200 RPM

Setting Of spindle RPM Press RPM key from front panel a RPM screen will be displayed than set the RPM by pressing UP/Down/Digit scroll key and enter to register the value. Than come out the RPM mode.

For starting and stopping of spindle rotation Press RPM key and than F1 for starting and F2 for Stopping

Temperature Setting:

In this parameter temperature of the bath is set from 30°C to 40°C

Press the TEMP key from the front panel a Temperature screen will be display than set the Temperature by using UP/Down/Digit scroll key and enter key to register the value. Than come out the Temperature mode. 

For Heater ON/OFF Press TEMP key and than F1 for Heater to be ON and F2 for Heater to be OFF.

Indication

If the temperature controller has started then the ONLED on the front panel and ETC-11L will glow.

If HEATER ON then the circulating pump will start by default

Run Time

To set the run time press TIME key from the front panel and set the time by pressing UP/Down/ Digit Scroll key and enter key to register the value. Than come out the time mode 

4.2.10  Lower the stirrer unit by pressing the down arrow key from the front panel. 

Press F1 Key (PREPARE) for starting of test, the temperature controller will be on.

4.2.11  To attain a constant temperature through out The vessel start the RPM with paddle attached to the spindle.

4.2.12  After getting the desired temperature at the lowest position of the stirrer unit (as per the method selected) the ready indication on the front panel will glow.

4.2.13  To View the individual jar temperature in between the test or prior to the test, from idle screen press F2 key to select jar probe.

Caution: If F1 key pressed to start the test and stirrer unit is not at the lowest position then an error indication will be display for 5 sec, Press down arrow key to park the stirrer unit at lowest position and rectify the error indicated.

4.2.14  To start the testing lift the stirrer unit and place the test sample in basket if using apparatus 1 and in jar if using apparatus2 than down the stirrer unit by pressing down key and start the test. 

5.0       Calibration:

5.1       CALIBRATION OF TEMPERATURE:

5.1.1    Set 37˚ C and measure the temperature using calibrated thermometer.

5.1.2    Of inside vessel at the following intervals : 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours.

5.1.3    Limit : The observed value should be between 36.5˚ C to 37.5˚ C.

5.1.4    Record the observations as given in the LOG BOOK.

5.2       CALIBRATION OF SPEED:

5.2.1    Set the speed as 50 RPM and measure the number rotations at the following time intervals :  30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours.

5.2.2    Limit: ± 4 % of the set speed.

5.2.3    Record the observations as given in the LOG BOOK.

5.2.4    In the same way, set the speed as 100 RPM and measure the number rotations at the following time intervals: 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours.

5.2.5    Limit: ± 4 % of the set speed.

   5.2.6    Record the observations as given in the LOG BOOK.

5.3       WOBBLING :

5.3.1    Check the shaft paddle wobbling using wobble meter at different time intervals as follows: 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours.  (Limit : ± 2 mm from vertical axis) Record the observations as given in the LOG BOOK.

5.3.2    Check the basket wobbling using wobble meter at the following time intervals as follows: 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours.   (Limit : ± 2 mm of maximum allowable run out at the bottom).

5.3.3    Record the observations as given in the LOG BOOK.

5.4       DISTANCE BETWEEN THE BOTTOM OF THE VESSEL AND THE BASKET:

5.4.1    Measure the distance between the bottom of the vessel and the basket. Limit: 25 ± 2 mm.

5.4.2    Record the observations.

5.5       CENTERING OF DISSOLUTION VESSEL:

5.5.1    Using the centering device check whether the dissolution vessel is at center.

5.5.2    If it is not at the center, then adjust its position to the center.

5.6       Calibration Frequency: Monthly for above parameter

5.7       Calibration with USP calibrators:

5.7.1    Use the following dissolution calibrators:

1.         Disintegrating type Prednisone tablets.

Use current lot of the above tablets for calibration.

5.7.2    To be prepared and use dissolution medium as per catalog given with USP calibrators tablets specification from the source.

5.7.3    To set all the parameters e.g. RPM, Apparatus 1 or 2 (Basket or paddle), temperature and time as per given in specification.

5.7.4    After completion of USP calibrator calibration calculate the percentage and compare with Acceptance Criteria as per given in specification.

5.7.5    Enter the details of calibration record in LOG BOOK

5.8              Calibration Frequency: Yearly for USP calibrator tablet

6.0                    Routine maintenance:

6.1                    Disconnect the instrument from power supply.

6.2                    Remove the water from tank. Clean the water tank with dry cloth and replace the fresh distilled water.

6.3                    Clean the Basket –Paddle and Bowl with water and than dry cloth.

6.4                    Clean the instrument with dry cloth.

SOP FOR OPERATION AND CALIBRATION OF NEPHELOMETER

1.0       Objective:

            The objective of this standard Operating Procedure is to lay down the procedure for

            Operation and Calibration of Nephelometer.

2.0       Scope:

            This SOP covers operating and calibration of Nephelometer .

3.0       Responsibility:

            Jr. Research Officer, Research Officer: Responsible for operation, calibration and  

            maintenance of the instrument as per procedure.

            Head of Department: Responsible for calibration and maintenance, timely as per               

            schedule.

            QA Officer/QA Manager: Review the records and governing the document.

4.0       Procedure:

4.1       Switch ON the instrument (back side switch).

4.2              Open the lid of the sample compartment & insert a test tube filled with the distilled water

            into the sample compartment then close the lid.

4.3       Rotate the switch of “set 0” to get “0” on LCD.

4.4       Open the lid, replace the test tube of distilled water by Formazine Std .Solution, and close

            the lid.

4.5       Rotate the switch of “Set 100” to get “100” on LCD.

4.6      Take the reading of sample in same way after setting standard.

4.7      Switch of the instrument.

Note:   Wash the test tube with distilled water after taking result.

5.0       Calibration Procedure:

            Solution 1:- Weigh accurately 5.0 g of Hydrazine Sulphate in 500 ml volumetric flask

            and add distilled water to make up to the mark. Leave the mixture to stand for 4 hours.

            Solution 2:- Weigh accurately 50.0 g of Hexam-Ethlenetetramine in 500ml volumetric

            flask and add make up the volume up to the mark with distilled water.

            Mix the equal volume of solution 1 and 2 form Formazine Turbidity Concentrate   

            Allow to stand in a closed container at 25º C to 30º C for 48 hrs to produce insoluble 

            while turbidity corresponding to 4,000 NTU (This solution is stable for 2-3 Months).

            Formazine Standard -100 NTU :- Dilute 25 ml of  the Formazine Turbidity Concentrate 

            to one liter with turbidity free distilled water to obtain 100 NTU Formazine Standard  for     

            setting 100 of  the instrument. (This Solution should be prepared weekly)

6.0              Routine Maintenance:

6.1              Clean the instrument with dry cloth.

6.2              Clean sample tube with methanol and than with water.

6.3              Clean the sample holder with dry cloth.