Category: QC SOP

SOP Title-Handling of Hazardous Chemicals

 

 

• OBJECTIVE

To lay down the procedure for handling of hazardous chemicals in Quality Control laboratory.

• SCOPE

This SOP is applicable to handling of hazardous chemicals in Quality Control laboratory.

• RESPONSIBILITY

3.1  QC Executive / designee shall be responsible for implementation of this procedure.

3.2   QC Head / designee shall be responsible for compliance of this procedure.

• PROCEDURE
• Handle the hazardous chemicals in the presence of QC personnel.
• Refer Material Safety Data Sheet (MSDS) before handling any chemical.
• Use acid proof hand gloves, safety goggles and other protective means.
• Carefully handle hazardous chemicals or solution preparations in fuming hood only.
• Maintain safe distance from chemical container, while pouring out or siphoning the chemicals.
• Use suction bulb or dispenser during transfer of solution..
• While preparing dilute Sulphuric acid take special care as follows:

First cool the aqueous solution in ice bath. Slowly add Sulphuric acid into solution with constant stirring. Never follow vice a versa procedure.

• Handle inflammable liquids and substances away from naked flame or electric spark.
• Clean any spillage immediately.
• Store all chemicals and solvent in specified area only.
• Store the poisonous chemicals in specified lock & key area.
• Spillage of hazardous chemicals should be avoided. Use spill kit to wipe out chemicals spillage.
• Unless otherwise specified, water should be poured on the surface of spillage to dilute the same.
• All necessary precautions for cleaning should be taken immediately by referring MSDS.
• List of hazardous chemical shall be maintained as per SOP.

 

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
QA	Quality Assurance
SOP	Standard Operating Procedure
No.	Number
°C	Degree Centigrade

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title-Handling of Analytical Samples

 

• OBJECTIVE

To lay down the procedure for Handling of Analytical Samples.

• SCOPE

This SOP is applicable to Handling of Analytical Samples in Quality Control laboratory.

• RESPONSIBILITY
• QC personnel shall be responsible for implementation of this procedure.
• QC Head /designee shall be responsible for compliance of this procedure.

 

• PROCEDURE
• In process sample inward register:

  On Receipt of Test request from Production Department, the QA Chemist shall provide the required quantity of sample and hand over to QC department. Concern sample receiver in QC shall be responsible for making entry in inward register and place the sample in the area assigned for it.

• Finished Product inward register

On Receipt of Test request from Production Department, the QA Chemist shall provide the required quantity of sample and hand over to QC department. Concern sample receiver shall be responsible for making entry in inward register and place the sample in the area assigned for it.

• Water sample inward register:

After sampling the water sample from the concern location, make entry in inward register  and place the sample in the area assigned for it.

• Validation Sample inward register:

 On Receipt of samples from the vendor or any other source . Concern QC person shall be responsible for making entry in inward register and place the sample in the area assigned for it.

• Miscellaneous sample inward register:

 On Receipt of samples from the vendor or any other source. Concern QC person shall be   responsible for making entry in inward register.

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
QA	Quality Assurance
SOP	Standard Operating Procedure.

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title-Dissolution Profile Comparison

 

• OBJECTIVE

To lay down a procedure  for  preparation of dissolution profile comparison protocol.

• SCOPE

This SOP is applicable for dissolution profile comparison  in quality control department.

 3.0 RESPONSIBILITY

• QC Executive – To follow the SOP.
• QC Head – Review, technical correction, training and monitoring of SOP.
• QA Head – Approval and implementation of SOP.

 

• PROCEDURE
• Dissolution profile comparison protocol shall be prepared and numbering shall be as follows;

QC/DPC/XX/YY

QC is the Quality Control

DPC is the Dissolution Profile Comparison

XX  is the sequential number of Protocol

YY is the calendar year

• The list of dissolution profile protocol shall be prepare as per define format.
• The dissolution profile protocol shall be prepared as per SOP.
• The dissolution profile protocol shall contain the following parameters;
  • Dissolution profile comparison protocol cover page
  • Protocol approval
  • Revision History
  • Objective
  • Scope
  • Responsibility
  • Procedure
• Calculation for (f₁) & (f₂)
• Acceptance Criteria
• Maintenance of raw data
• Deviation (if Any)
• Reference
• Procedure: Perform 12 dosage units dissolution as per analytical standard test procedure for test product and RLD sample.
• Calculation :  Calculate the difference factor (f₁) & similarity factor (f₂)

Where,

n = Number of Dissolution time points

Rt and Tt are the cumulative percentage of the drug dissolved at each of the selected n time points of the reference and test product, respectively.

• An aqueous medium with pH range 1.2 to 6.8 (ionic strength of buffers the same as in USP) should be used. To simulate intestinal fluid (SIF), a dissolution medium of pH 6.8 should be employed. A higher pH should be justified on a case-by-case basis and in general, should not exceed pH 8.0. To simulate gastric fluid (SGF), a dissolution medium of pH 1.2 should be employed without enzymes. The need for enzymes in SGF and SIF should be evaluated on a case-by-case basis and should be justified.

The dissolution time points for both the profiles shall the same (e.g. 15, 30, 45, 60 minutes). The reference batch used shall be the most recently manufactured product.

• To allow use of mean data, the percent coefficient of variation at the earlier time points (e.g., 15 minutes) shall not be more than 20%, and at other time points should not be more than 10%
• In case release % is more than 85% for intermediate release dosages forms. Then similarity factor & difference factor is not required.
• Acceptance criteria:

For curves to be considered similar, f₁ values should be upto 15 (0-15), and f₂ values  should be greater than 50 (50-100)..

• Additionally, All calculation for comparison data and graphical presentation of dissolution profile shall be prepared in additional sheet as a supporting data. (If required).

 

• ABBREVIATION(s)

Abbreviation	Full Description
SOP	Standard Operating Procedure
QA	Quality Assurance
QC	Quality Control
ARD	Analytical Research Department
RLD	Reference Listed Drug
FDA	Food and Drug Administration
USP	United States Pharmacopeia
SIF	Simulate Intestinal Fluid
SGF	Simulate Gastric Fluid

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title: Cleaning of Walk-In Type Stability Chambers

1. Objective:

To lay down the procedure for Cleaning of Walk-In Type Stability Chambers in laboratory.

2. Scope:

This SOP is applicable for Cleaning Walk-In Type Stability Chambers in Laboratory.

3. Responsibility:

• All Analysts /Scientists are responsible to follow this SOP.
• Senior Scientist/Designee is responsible to review and supervise compliance with this SOP.
• Head- QC is responsible to assure adherence to this SOP.
• Head- QA is responsible for approval of SOP and to ensure adherence to this SOP.
• Quality Assurance representative is responsible to distribute, retrieve and archive of this SOP.

4. Accountability:

Head – QC / Head – Quality Assurance

5. Procedure:

5.1	For External Cleaning
1	De-dust the chamber externally with a clean and dry cloth.
2	Clean the glass window externally with mild detergent solution.
3	Wipe the glass surface with clean dry cloth to remove trace of detergent.
4	Once a month remove the adhered dust by using mild non-abrasive detergent solution
	(Frequency : Once a month)
5.2	For Internal Cleaning
1	Login to the HMI Screen.
2	Select Door Access option on the HMI and Enter User Id and Password.
3	Note: Make entry in the “Entry and Exit Logbook” of the stability chamber.
4	De-dust he chamber with clean dry cloth.
5	Clean the glass window internally with mild detergent solution.
6	Wipe the glass surface with clean dry cloth to remove trace of detergent.
7	Clean the tray using mild non-abrasive detergent solution. Ensure that all corners are cleaned and free from dust.
8	Wipe the internal walls and tray with clean dry lint free cloth to remove any traces of detergent solution.
9	Once a month, mop the internal floor of the walk-in type stability chamber with mild non-abrasive detergent solution.
10	Note: Cleaning is to be carried out with doors closed.
11	Once cleaning is complete, make entry in the cleaning record.
12	Press Yellow colour knob on the right side of the door to exit.
13	Note: Make entry in the “Entry and Exit Logbook” of the stability chamber.

6. Definitions and Abbreviations:

• Abbreviations:

Abbreviation	Expansion
SOP	Standard Operating Procedure
HMI	Human Machine Interface

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title- Chromatographic Analysis and Documentation

 

• OBJECTIVE

To lay down the procedure for chromatographic analysis and documentation.

• SCOPE

This SOP is applicable for chromatographic analysis and documentation in quality control department.

 

• RESPONSIBILITY

3.1 QC Executive / designee shall be responsible for Implementation of this procedure.

3.2  QC Head / designee shall be responsible for compliance of this procedure.

 

• DEFINITION(s)

Chromatography is defined as a procedure by which solute are separated by a dynamic migration process in a system consisting of two or more phases, one of which moves continuously in given direction and in which the individual substances exhibit different mobilities by reason of differences in adsorption, partition, solubility, vapor pressure, molecular size, or ionic change density. The individual substances thus separated can be identified or determined by analytical procedure.

 

• PROCEDURE
  • Mobile phase
    • The mobile phase shall be prepared as per respective standard test procedure.
    • The mobile phase bottle should be labeled as per define format.
    • Use freshly prepared mobile phase for HPLC analysis which will be valid for one day only.
    • Use HPLC grade solvents for mobile phase preparation.
    • Always filter mobile phase through 0.45micron filter before use.
    • Do not use any hazy mobile phase during HPLC analysis.
  • Standard preparation
    • The standard preparation shall be prepared as per respective standard test procedure, taking into consideration the analyte stability and storage requirement.
  • Analysis
    • Always use fresh HPLC vials & septa for analysis.
    • Prepare the sample set as per respective standard test procedure.

 

• Prior to start the sample set analyst shall take the printout of sample set and shall make the sign with date on it. Reviewer shall also be review the sample set and put the sign and date. In case reviewer shall not be available in the shift, then another analyst shall review the sample set.
• A system suitability check shall be injected at the beginning to check the mobile phase preparation and saturation of HPLC column.
• Injection for system suitability check shall be processed, and shall kept with main sample set .
• The system suitability shall be established as per the standard test procedures, before proceeding the analysis.
• A printout of sample set has to be attached with analytical worksheet along with chromatograms.
• Injection sequence shall be as per the standard test procedure. If no specific sequence are mentioned in the standard test procedure then following the sequence flow as per below:

Blank →System suitability solution (if applicable) →Placebo (if applicable) →Impurity standard (if applicable) →StandardsàSamples →Bracketing Standard.

• In case No. of injections for standard are not mentioned in standard test procedure then inject the five replicates of the standard. In case where diluted standard has to be injected then inject six replicates for the same.
• The system suitability shall be demonstrated throughout the run by bracketing standards RSD.
• After every six injections or end of the sample set, a single standard (Bracketing standard) solution injection shall be made and the cumulative/overall RSD of these standards with earlier system suitability standards shall be calculated. The acceptance criteria for system suitability shall be overall RSD not more than 2.0 % for assay or as per standard test procedure.
• In case system suitability with bracketing standard not met with acceptance criteria in that case previous sample shall be invalidated with proper documents.
• Wherever system suitability fails in %RSD of replicate injections, following points shall be checked.

Injector / pump leak.

Column backpressure.

Temperature record of the working area.

Visual inspection of mobile phase and tubing for any possible air bubble.

Column saturation

• Wherever system suitability fails in case of tailing / column efficiency, regenerate / wash the column or replace the column and repeat the analysis. Submit the sample solution sequence only after passing the system suitability criteria.
• The retention time of standard and sample responses shall agree with in 10 % of actual retention time throughout the analysis for same concentration solutions.
• Resolution, tailing factor and theoretical plates shall comply with respective standard test procedure.

• Adjustment of Chromatographic Condition;
  • Composition of mobile phase: The amount of the minor solvent component may be adjusted by ± 30% relative to minor component or ±2% absolute, whichever is the larger, if required to meet the system suitability criteria. No other component is altered by more than 10% absolute.
  • pH of the aqueous component of the mobile phase: ±0.2 pH, unless otherwise stated in the standard test procedure, or ± 1.0 pH when neutral substances are to be examined.
  • Concentration of salts in the buffer component of a mobile phase: In the buffer component of a mobile phase: ± 10 per cent
  • Detector wavelength: No adjustment permitted
  • Stationary phase:
  • Column length: ± 70 %
  • Column internal diameter: ± 25 %
  • Particle size: Maximal reduction of 50 %, no increase permitted.
  • Flow rate: ± 50 per cent. When in a monograph the retention time of the principle peak is indicated, the flow rate has to be adjusted if the column internal diameter has  been changed. No decrease of flow rate is permitted if the monograph uses apparent  number of theoretical plates in the qualification section
  • Injection volume: May be decreased, provided detection and repeatability of the peaks(s) to be determined are satisfactory.
  • Gradient elution: the configuration of the equipment employed may significantly alter the resolution, retention time and relative retention time described in the method. Should this occur, it may be due to excessive dwell volume which is the volume between the point at which the 2 eluents meet and the top of the column.
• Gas chromatography
  • Stationary phase:

Column length : ± 70 %

Column internal diameter: ± 50 %

Particle size : Maximal reduction of 50 %, no increase permitted

Film thickness: -50 % to + 100 %

• Flow rate : ± 50 %
• Temperature: ± 10 %
• Injection volume :May be decreased, provided detection and repeatability are satisfactory.

• Recording of chromatograms
  • Prior to process the chromatogram, set the integration parameters (i.e. peak width, peak threshold, minimum height, minimum area) appropriately for proper marking of peaks.
  • All the chromatograms for establishment of system suitability and up to entire run shall be recorded and documented. Appropriate remarks (if required) shall be recorded on these by the analyst. Each chromatogram shall be duly signed off by the analyst and reviewed and signed by the reviewer.
  • The chromatograms which are disregarded and not considered for calculations shall be invalidated with reason and proper documentation. The reason for disregarding the chromatograms may be system suit check, abnormal peak shape, system suitability failure due to variation in area count/inconsistent area, faulty integration, ghost peak or any other reason.
  • The analyst performing the analysis shall write the reason for disregarding with “INVALID” stamp on the chromatogram. The disregarded chromatogram shall be filed along with the valid chromatogram.
  • During system suitability only initial replicate injections shall be considered for calculation of RSD.
  • Reprocessing of chromatograms, if necessary, shall be documented with reason(s) & “INVALID” stamp.
  • Any unknown peak found in the chromatogram other than the diluent, placebo and impurity peaks shall be integrate and considered as unknown peak.
  • Integration parameters such as peak width, slope sensitivity, minimum peak area / height etc. shall be recorded as used for each chromatogram or print out of integration parameters shall be attached with the chromatogram.
  • Attached the sequence print and method print for the entire analysis.
  • Sequence print shall contain Vial No, Tray No, Sample Name, Sample ID, Method File, Data File, Injection Volume.
  • Analyst and reviewer shall sign all the chromatograms with date.

 

• Enter the usage details in instruments, Columns usages record and reference / working standard usage logbooks.
• Affix the Mobile Phase bottle status label.
• Affix the Label For all Solution as per Annexure.

 

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
e.g.	Example
No.	Number
°C	Degree Centigrade
RH	Relative Humidity
%	Percent
N	Normal
DL	Detection Limit
QL	Quantitation Limit
ppm	Parts per million
ppb	Parts per billion
USP	United State of Pharmacopoeia

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title-Calibration of High Performance Liquid Chromatography (HPLC)

 

• OBJECTIVE

To lay down the procedure for Calibration of High Performance Liquid Chromatography.

 

• SCOPE

This SOP is applicable to Calibration of High Performance Liquid Chromatography Quality Control laboratory.

• RESPONSIBILITY
• QC personnel shall be responsible for implementation of this procedure.
• QC Head /designee shall be responsible for compliance of this procedure.
• PROCEDURE :-
•  Calibration Frequency: Calibrate the instrument quarterly or after major part replacement.

• HPLC is calibrated for following parameters:
•            Flow rate Accuracy of pump
•            Injection Volume Accuracy of the pump
•            Precision
•            Injector Linearity
•            Detector Linearity
•            Wavelength Accuracy
•            Carry over test
•            Drift and Noise
•            Gradient Proportional Value (GPV Test)
•            Oven calibration
•            Software Verification

PARAMETERS	LIMIT
Flow rate Accuracy of pump	± 2.0% from the set value
Injection Volume Accuracy of the pump	± 2.0% from the set value
Precision	RSD of Area 2.0%  and Retention time NMT 1.0%
Injector Linearity	Correlation Coefficient ≥ 0.999
Detector Linearity	Correlation Coefficient ≥ 0.999
Wavelength Accuracy	Maxima at 205 nm ±2 nm, Minima at 245 nm ± 2 nm and Maxima  at 272 nm ± 2 nm.
Carry over test	NMT 0.01 %
Drift and Noise	Drift ±5000 uV  and ± Noise 200 uV
Gradient Proportional Value   (GPV Test)	10 % Height (8.0 % to 12.0%) ,50% Height (45% to 55 %)  and 90.0% Height (85.0% to 95.0%
Oven calibration	± 2.0°C of set value
Software Verification	It should pass

• PUMP CALIBRATION:
•    Remove the column and purge the pump with Distilled water. Set the flow rate 0.5 ml/min. and allow to run for 5 min .Take dry volumetric Flask and note the weight (W₁). Collect the eluent carefully into the volumetric flask for exactly 5 minutes by noting the time using a calibrated stop watch. Note the weight (W₂) of the bottle with the collected eluent. Calculate the flow rate by the formula given below.  Repeat the procedure twice again. Similarly repeat the procedure with flow rates of 1.0 ml/min. and 2.0 ml/min twice  from all channels.

(W₂ -W₁)                1

Flow = —————– x ———— =    ————- ml/min.

0.9970                        5

 

Acceptance Criteria: The flow rate shall not vary more than 2.0% from the set value.  

 

• INJECTION VOLUME ACCURACY:   
•        Fill the HPLC vial with Purified  water and allow  vial to come at room temperature note  the  weight of filled vial in gm. Carry out 20 consecutive injection of 5 μl  and after the completion of the injection reweight the vial (in gm). Follow the same for 10μl, 20μl, 50 μl  using 10 consecutive Injection and  100 μl using 5 consecutive injection.

 

CHROMATOGRAPHIC CONDITION

Column                   : Octadecylsilyl silica gel (C18), 5 µ or Equivalent

Dimensions             : 250 x 4.60 mm

Wave length            : 272 nm

Flow rate                 : 1.0 ml/minute.

Injection Volume    : 5μl, 10μl, 20μl, 50μl &  100μl

Mobile phase          : Distilled Water

Run Time                : 1.0 min

Calculation :-

(W₁ – W₂) x 1000

Volume  =   —————————– =    ———- µl

0.9970   x      Y

W1      :           Initial weight of filled vial

W2      :           Initial weight of vial after 10 Injection

Y         :           No of Consecutive Injection

Density            : 0.9970 at 25⁰ C

• PRECISION :-

   Weigh accurately 30 mg of caffeine standard and transfer to a 100 ml volumetric flask, dissolve  and make  up to the volume with mobile phase. Further    dilute 10 ml of this above solution to 100 ml, mix and make up to volume  with mobile phase.  Inject the one Blank Preparation and Six standard preparation. Record the chromatograms.

 

CHROMATOGRAPHIC CONDITION

Column                         : Octadecylsilyl silica gel (C18) 5 µ or                                                            Equivalent

Dimensions                   : 250 x 4.60 mm

Wave length                  : 272 nm

Flow rate                       : 1.0 ml/minute.

Injection Volume          : 20 μl

Mobile phase                 : (Acetonitrile  : Water) (70: 30)

Run Time                      : 10 .0 min

 

•                 INJECTOR LINEARITY :            

Weigh accurately 30 mg of caffeine standard and transfer to a 100 ml    volumetric flask, dissolve  and make up to the volume with mobile phase. Further, dilute 10 ml of this above solution to 100 ml mix and make up to volume with mobile phase. Inject 5µl, 10µl,20 µl, 50 µl and 100 µl 3 times and plot the graph volume against mean area. Find out linearity coefficient.

 

CHROMATOGRAPHIC CONDITION :

Column                       : Octadecylsilyl silica gel (C18) 5 µ or Equivalent

Dimensions                 : 250 x 4.60 mm

Wave length                : 272 nm

Flow rate                     : 1.0 ml/minute.

Injection Volume        : 20 μl

Mobile phase               :(Acetonitrile  : Water) (70: 30)

Run Time                    :10.0 min

 

•  DETECTOR LINEARITY :          

Weigh accurately 30 mg of caffeine standard and transfer to a 100 ml volumetric flask, dissolve  and make up to the volume with mobile phase. This solution is (Stock A) further Dilute.

2 ml of the stock A to 50 ml of volumetric flask mix and make up to volume with mobile phase i.e. concentration of 12 ppm.

4 ml of the stock A to 50 ml of volumetric flask mix and make up to volume with mobile phase i.e. concentration of 24 ppm.

6 ml of the stock A to 50 ml of volumetric flask mix and make up to volume with mobile phase i.e. concentration of 36 ppm.

8 ml of the stock A to 50 ml of volumetric flask mix and make up to volume with mobile phase i.e. concentration of 48 ppm.

10 ml of the stock A to 50 ml of volumetric flask mix and make up to volume with mobile phase i.e. concentration of 60 ppm.

 

CHROMATOGRAPHIC CONDITION :

Column                       :Octadecylsilyl silica gel (C18) 5 µ or Equivalent

Dimensions                 : 250 x 4.60 mm

Wave length                : 272 nm

Flow rate                     :1.0 ml/minute.

Injection Volume        :20 μl

Mobile phase               : (Acetonitrile  : Water) (70: 30)

Run Time                    : 10.0 min

 

•   WAVELENGTH ACCURACY :-

Weigh accurately 30 mg of caffeine standard and transfer to a 100 ml volumetric flask, dissolve  and make  up to the volume with mobile phase. Further, dilute 10 ml of this above solution to 100 ml mix and make up to volume  with mobile phase.  Inject the one Blank Preparation and Six standard preparation. Record the chromatograms.

CHROMATOGRAPHIC CONDITION :

Column                       : Octadecylsilyl silica gel (C18) 5 µ or Equivalent

Dimensions                 : 250 x 4.60 mm

Wave length                : 201 nm to 209 nm, 241 nm to 249 nm and 268                                           nm to 276 nm.

Flow rate                     : 1.0 ml/minute.

Injection Volume        : 20 μl

Mobile phase               : (Acetonitrile : Water) (70: 30)

Run Time                    : 10.0 min

 

•  CARRY OVER TEST :-
•         Weigh accurately 30 mg of caffeine standard and transfer to a 100 ml volumetric flask, dissolve  and make  up to the volume with mobile phase.

CHROMATOGRAPHIC CONDITION :

Column                            : Octadecylsilyl silica gel (C18) 5 µ or                                                             Equivalent

Dimensions                    : 250 x 4.60 mm

Wave length                     : 272 nm

Flow rate                          : 1.0 ml/minute.

Injection Volume             : 20 μl

Mobile phase                    : (Acetonitrile  : Water) (70: 30)

Run Time                         : 60.0 min

 

• GRADIENT PROPIOTINAL VALVE :-

CHROMATOGRAPHIC CONDITION :

Pump A                       :  0.3 % acetone in distilled water.

Pump B                       :  Distilled water.

Flow rate                     :  1 ml/min.

Wavelength                 :  254 nm.

Column                       :  No column, but union guard column directly                                        connected to the detector.

B.CONC                     :  100 %

Time Programmer :-

Time in min.	Function	Conc. in %
0.01	B.Conc	90.0
10.00	B.Conc	90.0
10.01	B.Conc	50.0
20.00	B.Conc	50.0
20.01	B.Conc	10.0
30.00	B.Conc	10.0
30.01	B.Conc	0.0
40.00	B.Conc	0.0
40.01	B.Conc	100.0
50.01	Stop	–

Where, A10, A50, A90, A100 in the graph, are the distance from the base line to the I, II, III, IV step in centimeters respectively.

Specification: Pump A concentration ratio of 10%, 50% and 90 %.

Precautions for Measurement:

Ensure that the mobile phase is degassed.

Open the drain valve and wash the flow line for 5 minutes, with the flow rate set 5 ml / min and B.CONC parameter to 100 %.

Then set the flow rate to 3 ml / min, close the drain valve and wash the flow line for 20 minutes.

Last, set the flow rate to 2 ml/ min and the B.CONC parameter to 100 and check that the baseline has become stable, then start measurement.

 

• Oven Calibration:
•  Carry out Oven Cabinet calibration at  30°C, 40°C, 50°C and 60°C with the help of suitable calibrated thermometer and record the observation.

 

• Software Verification
• Go to the tools bar and select “Tools”, than select “Check program files”, a window “Check the program files” will display.
• Click on execute and wait for few seconds.
• Now click on Browse & than take the report print. All the parameter of software verification should be pass.
• Calibration report shall be prepared as per Annexure.
• After the successful completion of calibration. Calibration label shall be affixed on the instrument.

 

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
QA	Quality Assurance
SOP	Standard Operating Procedure.
µl	Micro liter
µm	Micrometer
%	Percentage
°C	Degree Centigrade
RSD	Relative standard Deviation
mv	milli volt
GPV	Gradient Proportional Valve

 

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title-Calibration of Gas Chromatography

 

 

• OBJECTIVE

To lay down the procedure for Calibration of Gas Chromatography.

• SCOPE

This SOP is applicable to Calibration of Gas Chromatography in Quality Control laboratory.

• RESPONSIBILITY
• QC personnel shall be responsible for Implementation of the procedure.
• QC Head /designee shall be responsible for compliance of this procedure.

 

• CALIBRATION PROCEDURE
  • Calibration Frequency:
    • Every six months and after critical parts replacement.
  • Chromatographic condition:

Column                       :           ZB 624, Length 30m, Inner Diameter                                          0.53mm, Film Thickness 3.0µm (or equivalent)

Detector                      :           Flame Ionization Detector

Carrier Gas                  :           Nitrogen gas

Flow rate                     :           2 ml/min

Injection temperature  :           150°C

Oven temperature       :           100°C

FID temperature         :           200°C

Injection volume         :           1 µl

Split ratio                    :           1:20

Run time                     :           10 min.

• Head space sampler parameter:

                        Incubation /vial temperature   :           80°C

Syringe temperature                :           85°C

Injection volume                     :           1 ml

Incubation time                       :           15 min.

• Standard stock solution:
  • • 5 % v/v Acetone solution in water:-

                 5ml of Acetone dissolve in 100ml of water.

    • 5 % v/v Tetrahydrofuran (THF) in water:-

                5ml of THF dissolve in 100ml of water.

  Standard solution A:

Take 5 ml Acetone standard stock solution and 20 ml Tetrahydrofuran standard stock solution in 200 ml with water.

              Standard solution B:

    Take 5 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution C :

           Take 10 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution D:

       Take 15 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution E:

Take 20 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

• Precision:
  • Pipette out accurately 1 ml of standard solution C into six separate 20 ml HS vial

5.5.2     Seal all the vials properly using septa.

5.5.3  Procedure:   Inject the blank vial (system blank) and six standard vials and again blank vial (for carry over) as per the procedure and calculate the RSD for Retention times and peak areas response ratio of individual peaks.

5.5.4                The relative standard deviation of area response ratio for acetone with respect to tetrahydrofuran for six replicate injections should not be greater than 5.0 %.

5.5.5   The relative standard deviation of the retention time for acetone with respect to tetrahydrofuran for six replicate injections should not be greater than 1.0 %.

5.5.6   Carry over: NMT 0.1 % (Blank after six injection/ Average peak area)

 

5.6 Linearity:

5.6.1   Pipette out accurately 1 ml of standard solution A into two separate 20 ml HS vial and seal the vials properly using septa. Similarly prepare two vial standard solution B, two vial standard solution C, two vial standard solution D and two vial standard solution E.

5.6.2    Inject all the vials, one by one.

5.6.3    Record the chromatogram of each injection and calculate the average area response ratio for acetone with respect to tetrahydrofuran.

• Draw a graph with injection volume on X-axis and average area response ratio on Y-axis.
• Curve should be linear and correlation co-efficient should not be less than 0.999.

5.7  Calibration of Column Oven Temperature Test

5.7.1   Take the first temperature reading from the thermometer and record the temperature.

5.7.2     Set the oven temperature to 100°C, 150°C and 230ºC and wait for at least 5 minutes after the oven goes ready for temperature stabilization

• Take the first temperature reading from the thermometer and record the temperature.

5.7.4                Acceptance Criteria :-

Difference at 100°C should be not more than 3°C.

Difference at 150°C should be not more than 3°C

Difference at 230°C should be not  more than 3°C.

• Software Verification

• Go to the tools bar and select “Tools”, than select “Check program files”, a window “Check the program files” will display.
• Click on execute and wait for few seconds.
• Now click on Browse & than take the report print.

Acceptance criteria: It should “Pass”

• Calibration Report of Gas Chromatography shall be prepared as per Format.
• After the successful completion of calibration. Calibration label shall be affix on the
• If the Calibration parameters are out of limit affix the Out of Calibration label and Inform to QC Manager/Designee.
• Record shall be maintained in instrument usage logbook.

 

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
QA	Quality Assurance
SOP	Standard Operating Procedure.
µl	Micro liter
µm	Micrometer
%	Percentage
°C	Degree Centigrade
RSD	Relative standard Deviation
GC	Gas chromatography
No.	Number
RT	Retention time

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title: Operation, Cleaning and Calibration of  Melting Range Apparatus

1. Objective:

To lay down the procedure for Operation, Cleaning and Calibration of Melting Range Apparatus.

 

2. Scope:

This SOP is applicable for Operation, Cleaning and Calibration of Melting Range Apparatus located in Quality Control Laboratory.

 

3. Responsibility:

• Quality Control: To prepare and review the SOP. To follow the procedures laid down for Operation, Cleaning and Calibration of Melting Range Apparatus as per this SOP.
• Quality Assurance Department: To review and approve the SOP and Annexures.

 

4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

 

5. Procedure:

 

5.1	Safety Precautions/Instructions:
5.1.1	Ensure all the electrical connections are properly earthed. Ensure the power plug of the   Melting Range is fixed in the socket properly.
5.1.2	Wear personal protective equipment’s such as safety glasses, hand gloves during operation of instrument.
5.1.3	Ensure that the ON/OFF switch is working properly by switching “ON” the Melting Range Apparatus.
5.1.4	Do not operate the instrument in the presence of flammable gasses or Liquid.
5.1.5	Switch “ON” the Instrument the 15 to 20 min. before the Start of analysis.
5.1.6	Cleaning of the furnace should be done at ambient temperature. Only surface of the furnace is recommended to clean with damp cloth. Do not use organic solvent for cleaning the furnace surface.
5.1.7	While Sample analyzing using capillary preparation, 3 capillaries should be filled about 3 to 5 mm of sample.
5.1.8	For sample whose melting point is not known, use heating RATE: 6.1 to 12.1 °C/MIN.
5.1.9	For ‘Heat & Cool’ Mode, it is recommended to use heating rate 1°C/MIN.
5.1.10	It is recommended to use ‘MANUAL’ operation for coloured samples.
5.1.11	For samples, that shrinks or become transparent during heating process, increase sample size and use heating rates: 6.1 ~ 12.1 °C/MIN.
5.1.12	Ensure the round Cover-slip on the Furnace Lid is clear (not foggy),
5.1.13	Ensure the surface of the Furnace should be clean from any foreign particles for a good thermal contact of the slide.
5.1.14	The presence of foreign particle on slide/cover slip will create the thermal difference and leads to erroneous results.
5.1.15	Ensure that there should not be Direct Air Drift in the furnace direction.
5.2	Front Panel-Keyboard:
5.2.1	Introduction to the soft Keys of Front Panel :
5.2.2	Left hand side keys on the front panel of the instrument are alphanumeric keys having dual functions are (A-Z, 1-9), Zero (0) with Dash (-), Dot (.), Slash (/), Decimal point (.) and Clear Entry (CE) Key.
5.2.3	Right hand side keys are functional keys as follows:
5.2.4	RESET- To Reset instrument.
5.2.5	BACK- To go backward for previous function display screen.
5.2.6	ESCAPE- To go back directly to main menu.
5.2.7	ENTER- To acknowledge the entered value/parameter.
5.2.8	LAMP- To Switch the Lamp ON/OFF.
5.2.9	STOP-In RUN-MANUAL mode, to hold isothermal condition, while Furnace temperature is raised with the user specified Heating rate.
5.2.10	LOCK- To lock the Start and End Temperature of Melting process in MANUAL mode.
	Sample   Placement:
5.3	Operation  of    Melting Range Apparatus:
5.3.1	Check the validity of calibration before starting any operations.
5.3.2	Ensure that the instrument is clean before operation.
5.3.3	Connect the power cord to 230 VAC/50 Hz stabilized power supply.
5.3.4	Switch ON the mains of   Melting Range Apparatus from back panel.
5.3.5	After instrument is powered “ON”. Press the “RESET” key, it gets initialized and the two line LCD display will read as follows
5.3.6	After 2 to 3 seconds screen will display the Main Menu: 1)Prog    2)Run   3)Print         4)Config    T° C: XX
5.3.7	1) ‘Program’         :To Add(create)/,edit, view, copy, delete Programs. 2) ‘Run’                 :To Run the desired Program. 3) ‘Print’               :To obtain print copy of Program, Result, Calibration               Report, Graph.  4) ‘Config’           : To set Date & time and Data transfer to PC/Printer.
5.3.8	T° C: The screen will display furnace Temperature of Instrument.
	1) For Addition of New Program:
5.3.9	Select   ‘Prog’ from main menu option by pressing ‘1’. press “ENTER” and the display will show : 1)ADD      2)VIEW 3) COPY   4) DELETE
5.3.10	To create program number 1 press‘1’ and press  “ENTER” and  the instrument will display as: Prog No(1-20): ADD/EDIT Mode
	NOTE: Instrument can save upto 20 numbers of program
5.3.11	Press “ENTER” and the display will read : Sample Name:_______
5.3.12	Use Alphanumeric Keys to Enter Sample Name. For Example:- Paracetamol
5.3.13	Press “ENTER” and the display will read as : Sample Preparation: _ 1) Slide 2) Capillary Slide Preparation: Analysis is done by using Slide. Capillary Preparation: Analysis is done by using Capillaries.
5.3.14	After Selection of sample preparation mode, Press “ENTER” and the display will read as : Method: _ 1) Stand      2) USP-1a Standard method: Need to set parameters Externally. USP-1a method: Method is set by default as per USP
5.3.15	Press ‘1’ for Stand or press ‘2’ for USP-1a  and press ‘ENTER’ the display will read as:
	NOTE: In method USP-1a the instrument will run with heating rate 1°C/Minute once the furnace temperature is reached to below 5°C of set temperature. Example: Set Temperature is 100°C then heating will start at 1°C/Minute once it reaches to 95°C.
	I) Standard Method
5.3.16	Mode: _ 1) Heat 2) Heat & cool After Selection of standard method the display will read as:   Mode: _ 1) Heat 2) Heat & cool   Heat mode: AUTO / MANUAL detection of Melting Point can be used with this mode Heat & Cool mode: Only manual detection of Melting Point can be done by this mode.
5.3.17	Press ‘1’ for Heat and press ‘2’ for Heat & Cool mode as per requirement
	NOTE: If ‘Heat & Cool’ Mode is selected, the Only ‘MANUAL’ detection is possible since the User has to Lock the Start & End temperature by pressing the LOCK key.
	Heat Mode:
5.3.18	In Heat mode the display will read as : Operation: _ 1) Auto 2) Manual Auto Operation:     Auto operation used in case of known sample/Colorless sample. Manual Operation: Manual operation is mostly used for coloured sample/ unknown sample.
5.3.19	Press ‘1’ Auto press ‘2’ for Manual operation as per requirement and press ‘ENTER’
	Auto Operation
5.3.20	While operation with Auto Mode the display the will read as: Set Temp ° C: __
5.3.21	Set Temperature is generally melting point of the Sample
5.3.22	After setting of temperature press Enter, the display will read as: Rate:    °C/MIN Please enter RATE
5.3.23	Enter the desired temperature at which it should rise the temperature in °C/min. The temperature will rise at specified desired rate when it reaches   10° C below the set temperature For Example: Suppose X sample set temperature is 100°C and rate of temperature increment is 1°C/ min then the temperature will rise    1°C /min only after it has reached to 90°C.
5.3.24	After selection of Rate press ‘ENTER’. The programme will get saved as a 1 number programme.
5.3.25	If set Temperature is not in the limit the display will read as: Invalid Entry RANGE: 8 to 350
5.3.26	Re-enter the valid Set temperature to continue the program.
	NOTE: The Furnace Temperature Cut-off Limit will be automatically calculated as Set Temp +15 °C for Heating Rates 0.2 to 6.0°C/Min. Example: Set Temperature is 100°C then Furnace Temperature Cut-off Limit will be 115°C.
5.3.27	For Heating Rates 6.1 to 12.1°C/Min, the by default cut-off limit is at 365°C.
	Manual Operation
5.3.28	While adding program with manual operation the display the will read : Set Temp ° C: __
5.3.29	After setting of the Temperature press ‘ENTER’ the display will read as: Furnace temperature Cutoff Limit: __
5.3.30	The Furnace temperature cut-off limit is the limit at which the instrument run will get abort. The cut off limit should be about 10°C more than the sample Melting point.
	NOTE: If the start and end temperature of melting are not locked manually, then the furnace will heat till CUT-OFF Temperature. ‘MANUAL’ operation recommended to use for colored/unknown samples.
5.3.31	After the selection of the furnace temperature the display will read as: Rate: __ °C/MIN Please enter RATE
5.3.32	After selection of Rate press ‘ENTER’. The programme will get saved as a 1 number programme.
5.3.33	If the Furnace Temperature Cut-off limit is less than SET Temperature or more than 365°C then the instrument gives an error message as below, Invalid Entry!…. Limits:  AB + 5 – 365 °C Where AB is set temperature value.
5.3.34	Re-Enter the Furnace Temperature Cut-off limit to continue the program.
	Heat & Cool Mode:
5.3.35	While adding Program with Heat & cool Mode  the display will read as: Set Temp ° C: __
5.3.36	After selection of the required temperature press ‘ENTER’ the display will read as: Furnace temperature Cutoff Limit: __
5.3.37	Rate: __ °C/MIN Please enter RATE   Select the Furnace  temperature cut-off Limit and press ‘ENTER’ , the display will read as:
5.3.38	After selection of Rate press ‘ENTER’. The programme will get saved.
	II) USP-1a Method:-
5.3.39	While adding Program with USP-1a Method (From Point no.5.3.14)
5.3.40	After Selection of USP-1a  method press ‘ENTER’ the display will read as : Operation: _ 1) Auto 2) Manual
5.3.41	Press ‘1’ for Auto and press ‘2’ for Manual operation as per requirement. And press ‘ENTER’
	i) Auto Operation
5.3.42	While operation with Auto Operation the display the will read as : Set Temp ° C: __
5.3.43	Enter the required temperature and press ‘ENTER’ to save the programme.
	ii) Manual Operation
5.3.44	While operation with Manual Operation the display the will read : Set Temp ° C: __
5.3.45	After selection of set temperature the display will read as: Furnace temperature Cutoff Limit: __
5.3.46	The instrument will set by default cut-off limit above 15°C of Set temperature OR can be set Cut-off limit manually as per requirement.  Press ENTER to save the programme.
	Note:  If program parameters are already stored at a particular program number then, to modify the desired parameters, follow the same procedure as mentioned from point number 5.3.9 to 5.3.32 for modification of existing program. The previously stored parameters are displayed, which may be acknowledged by pressing ‘ENTER’ or may be modified as per requirement.
	I) EXPLANATION OF MODE :
5.3.47	Modes are two types i.e. Heat Mode and Heat & Cool Mode
	HEAT MODE:
5.3.48	In Heat mode, the furnace will heat till the end temperature of melting range is detected or till the Furnace cut-off temperature is attained.
	HEAT & COOL MODE:
5.3.49	Only MANUAL operation is to be carried out with this mode
	II) EXPLANATION OF OPERATION :
5.3.50	Operation are of two types i.e. ‘Auto operation’ and ‘Manual operation’
	AUTO OPERATION:

 

5.3.51	The lamp will automatically adjust to optimum intensity depending on the brightness of the sample, in standard mode, below 5°C of the SET temperature & in USP-1A mode, 10°C below the set temperature. The system will then continue to heat with SET RATE. The start temperature will be automatically logged in, on the basis of slope changes. The end point is also detected automatically to give the melting range for the particular sample.
	2) Procedure to RUN  Programme :
5.3.52	Before Running the instruments, check the cleanliness of the instrument.
5.3.53	To Run the instrument, press ‘2’ from main menu 5.3.6  and the display will show : Prog No.: _ Inst. Calibrated
5.3.54	For Example To run program no. 4.
5.3.55	Enter Program No. and Press “ENTER” and the display will read as : Prog No.: 4      R_N0.: 1 1) Run 2) View: _
	NOTE: R. No. indicates the run number for particular day.
5.3.56	If entered  program no. does not exist then display will read as: Prog No.:4 Prg does not exist!.
5.3.57	If program does not exists then display will returns to the main menu
5.3.58	Press ‘2’ i.e. View to check the Program details.
5.3.59	To Run the program press ‘1’ i.e. Run and press “ENTER” the display will read as : Sample No: _
5.3.60	Enter Sample Number/Batch Number if it is available or else keep it as 1 number  by using alphanumeric keys and Press “ENTER” and the display will read as: Id. No.:_
	NOTE: Maximum 10 character sample no. and I.D can enter.
5.3.61	Enter Id. No. by using alphanumeric keys and Press “ENTER” to run the program. Furnace will start heating.
5.3.62	The furnace temperature start rising fast till the Sample Insertion Temperature arrives. Note:- The sample insertion temperature is the temperature below 15°C of set temperature. If the set Temperature is below 100° C then the sample insertion temperature will be 10° C   below the set temperature. Example: If SET Temperature is 114° C then the Furnace Temperature will rise fast till 99.0° C
5.3.63	The display will read as: T°C: 26.5 SET: 114.0 Wait Heating! 99.0
	NOTE: The furnace Lid should be placed on the furnace before start the analysis.
5.3.64	If furnace temperature is more than the Sample Insertion temperature, then the furnace cooling fan will turn “ON” internally until its temperature reaches below sample insertion temperature and the display will read as: T°C: 100.1 SET: 114.0 Wait cooling! 99.0
5.3.65	After the Furnace reaches Sample Insertion Temperature the instrument will give beep sound and the display will read as: T°C: 99.0 SET: 114.0 Keep Sample & ‘ENTER’
	NOTE: If ‘ENTER’ key is not pressed within 5 minutes, furnace will cool down to room temperature for the purpose of power saving and safety.
5.3.66	Prepare sample as per required method i.e. Slide and Capillary method
	Sample Preparation for Slide method:
5.3.67	Required Accessories for Slide method 1)      Clear Glass slide 2)       Clear Round Cover slip 3)       Clear dry mortar and pastel 4)       Small Spatula 5)       Finely crushed sample powder.
5.3.68	SAMPLE POWDER 22 DIA. COVER SLIP MICRO –SLIDE 1 MM THICK
5.3.69	Place the finely powdered sample on the one side of the glass slide. Finely spread the sample with the help of spatula. Place the round cover slip on the sample. Check for the sample is homogeneously spread in a circle.
	Sample Preparation for Capillary method:
5.3.70	Required Accessories for Slide method 1)      Clear dry mortar and pastel. 2)      One sealed Capillary tube 3)       Sample Filling Tube 4)      Tissue paper
	NOTE: Place a round cover slip for Slide method & square cover slip for Capillary method on the furnace lid. In between the Runs, the furnace lid should be sufficiently cooled.
5.3.71	Put the sample and press ‘ENTER’  the display will read as: T°C: 104.5 SET: 114.0 Heating! 109.0
5.3.72	The furnace will heat up to 5 °C below Set temperature and the Lamp will be switched ON automatically as soon as the Furnace temperature reaches 5°C below Set Temperature and the display will read as: Switching Lamp On…
	NOTE: i.      Instrument checks the light level & adjusts the illuminator intensity automatically for the optimum sensitivity level of Photo sensor. ii.    In AUTO operation, if the sample is not placed properly / sample quantity is less / sample preparation is not correct, a message will display accordingly error messages .
5.3.73	Now the Furnace temperature rises as per specified rate and the display will read as: T°C: 109.0 ST°C: ____ AUTO ET°C: ____
	NOTE: For MANUAL operation, ‘MANUAL’ will appear in left corner of the display instead of ‘AUTO’
5.3.74	While Program running with AUTO operation, when the sample start melting, the photo sensor will detects start of the melting temperature and will record it automatically and display will show recorded temperature as “ST” as shown below: T°C: 114.5 ST°C: ____ AUTO ET°C: ____
5.3.75	Furnace temperature will rise as per the specified Heating Rate. When the sample melts completely instrument will record the end temperature i.e. ‘ET’ and the display will read as: T°C: 115.5 ST°C: 114.5 AUTO ET°C: 115.5
	NOTE: While sample running with MANUAL operation Start temperature ‘ST’ and End temperature ‘ET’ is recored manually by using ‘LOCK’ key.
5.3.76	After completion of melting point, press ‘ESC’ the display will return to Main menu.
	3)  Procedure to Print  Reports and programmes :
5.3.77	To Print the data like Program, Result, Calibration Report, Graph, press ‘3’ from main menu refer point number 5.3.6 The display will show : 1) View Prev.Result 2) Print: _
5.3.78	Press ‘1’ and press ‘ENTER’, to view the previous result. The Display will read the complete Set of parameters and Results in Screen by screen,
5.3.79	Press ‘2’ and press “ENTER” to obtain a hard copy of Program, Result, Calibration Report, and Graph the display shows : 1) Prg Para 2) Rpt: _ 3) Cal Rpt  4) Graph
5.3.80	For the hardcopy of  last RUN report Press ‘2’ and ‘ENTER
5.3.81	1) Select 2) All: _ To print Hardcopy of program parameter  Press ‘1’ & press “ENTER” the display will show :
5.3.82	Select ‘1’ to print the hard copy of single selected Program parameter and select ‘2’ to print hard copy of all saved program parameter.
5.3.83	Prog No. (1-20): To print the hard copy of single program press ‘1’ and ‘ENTER’, the Display will read as:
5.3.84	Select the required program number and press ‘ENTER’.
5.3.85	3) Cal Rpt:-To print the Instrument Calibration report done in Factory press ‘3’ & ‘Enter’.
5.3.86	To print the hardcopy of last run sample graph press ‘4’ & ‘Enter’.
	Note: The ‘Graph’ option will be displayed only if last sample RUN was performed in AUTO operation. No graph is available for MANUAL Operation and serial transfer mode.
5.3.87	While Printing data  the display will show: PRINT MODE…
5.3.88	While printing is in Progress the display will show: WAIT… PRINTING DATA and after printing the display will show below screen and will return to main menu PRINTING OVER…. If printer is not connected then the display will show below screen and will return to main menu. PRINTER NOT READY!.
	NOTE: Only Last run result can be viewed or printed.
	Note: For invalid Entry, the entry position gets blank/clear and cursor returns to start of the entry
5.4	Calibration Procedure:
	Frequency:  Every Quarterly or after carrying out any major maintenance work.
5.4.1	Generate the A. R. No. on the “Instrument/Equipment Calibration Register” as per SOP and give requisition to QA department for calibration formats of instruments on the due date of calibration.
5.4.2	For issuance of calibration report refer SOP “Control of documents, Preparation, Approval, Issuance and Maintenance.
5.4.3	Perform the calibration of instrument as per Annexure No. 01 and procedure mentioned below.
5.4.4	Determine the melting points of the following standard materials: 1) Vanillin 2) Sulphanilamide 3) Caffeine 4) Benzoic Acid
5.4.5	Follow the same procedure from point no. 5.3.52 to 5.3.76
5.4.6	Enter the details in calibration report Annexure No. Ensure that the results obtained, comply with the acceptance criteria.
5.4.7	Update the “Instrument/Equipment Usage Log Book”, and “Calibration Schedule” as per SOP “Calibration of Instruments and Equipments in Quality control”. Update the “Annual Calibration Schedule” as per SOP,“Equipment’s / Instruments Calibration”
5.4.8	Affix the “Calibration Status Label” on the Instrument as per SOP “Calibration of Instruments and Equipments”.
5.4.9	If calibration activity is outsourced, the values are to be transcribed in the Annexure No. 01, and external party certificate to be attached.
5.4.10	For Out of calibration/breakdowns, follow procedure as per SOP “Calibration of Instruments and Equipments”.
5.5	Cleaning Procedure:
5.5.1	Before cleaning the instrument, unplug the power cord from the power source.
5.5.2	It is good laboratory practice to wipe away any spillage with dry cloth.
5.5.3	The exterior surface can be cleaned with soft dry cloth.
5.5.4	Do not use solvents like Chloroform, toluene.
5.5.5	Before and After use, clean the apparatus by removing sample particles powder from the platform using a brush and finally clean with dry cloth.
5.5.6	Use damp cloth or tissue paper to clean the lens. Do not adjust camera aperture since it is factory setup. The lens cleaning activity is recommended weekly. Always put lens cap when instrument is not in operation. User should not try to remove camera from Instrument since its alignment is factory adjusted.

 

6. Definitions / Abbreviation:

• Definitions:
• Calibration: Tests performed to ensure that the instrument/equipment is functioning as per set parameters. These tests are performed as per predefined frequency and also after any major repair or replacement of parts.
• Melting point: The temperature at which the solid and liquid forms of a pure substance can exist in equilibrium.
• Melting Range: The temperature range from which the crystals of the substance
• Principle: Melting point is the temperature at which a solid becomes a liquid at standard atmospheric pressure; at this point, solid and its liquid are in an equilibrium state at a certain pressure. Melting point is one of the physical properties of a compound by which it is identified.
• Abbreviation:

Abbreviation	Expansion
SOP	Standard Operating Procedure
QC	Quality Control

QA	Quality Assurance
A. R. No.	Analytical Reference Number
° C	Degree Celsius
MIN	Minutes
Temp	Temperature

Calibration Report

Sr. No.	Standards	Acceptance Criteria	Observed Values	Complies / Does Not Complies
1.	Vanillin	81°C – 83°C
2.	Benzoic Acid	121°C -123°C
3.	Sulphanilamide	163°C – 165°C
4.	Caffeine	234°C – 237°C

 

Remark: The result Complies/Does not complie’s as per acceptance criteria.

 

Performed By: ________________     Date: ____________              Checked By: ______________          Date: ____________

                                                       

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

SOP Title: Operation, Cleaning and Calibration of Digital Viscometer

1. Objective:

To lay down the procedure for Operation, Cleaning and Calibration of Digital Viscometer.

 

2. Scope:

This SOP is applicable for Operation, Cleaning and Calibration of Digital Viscometer located in Quality Control Laboratory.

 

3. Responsibility:

• Quality Control: To prepare and review the SOP. To follow the procedures laid down for operation, cleaning and calibration of Digital Viscometer as per this SOP.
• Quality Assurance Department: To review and approve the SOP and Annexures.

 

4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

 

5. Procedure:

 

5.1	Safety Precautions/Instructions:
5.1.1	Ensure all the electrical connections are properly earthed. Ensure the power plug of the Digital Viscometer is fixed in the socket properly.
5.1.2	Wear personal protective equipment’s such as safety glasses, hand gloves during operation of instrument.
5.1.3	Turn the power switch to the ON position and allow the viscometer to warm up for 10 minutes before performing auto zero.
5.1.4	The user should ensure that the substance placed under test do not release poisonous, toxic or flammable gases at the temperature which they are subjected to during the testing.
5.2	Instrument Operation:
5.2.1	Introduction to the front screen of Digital Viscometer:
5.2.3	Status Bar- The Status Bar provides information relating to the ate and time and various connection to the Viscometer.
	Test Name- The Test Name identifies the name of the currently loaded test.
	Title Bar-The tittle bar identifies the activity to be conducted in the current view.
	Data Fields-The data field includes measurement results and test parameter.
	More/Less Bar-  The More/Less Bar informs the user when more Data Fields are available and when the number Data field can be reduced.
	Command Keys-The command Keys indicate action that can be taken. These Keys will vary from view to view depending on what action are relevant.
5.3	Operation of Digital Viscometer:
5.3.1	Check the validity of calibration before starting any operations.
5.3.2	Ensure that the instrument should be cleaned before operation.
5.3.3	Connect the power cord to 230 VAC/50 Hz stabilized power supply.
5.3.4	Switch ON the mains of Digital Viscometer.
5.3.5	Allow 10 minutes warm up time before operation of Digital Viscometer.
5.3.6	After instrument is powered “ON”. The Viscometer Present Blue screen followed by auto zero screen. Remove the shipping cap before auto zero.
5.3.7	AUTO ZERO:
5.3.8	The Auto Zero should be perform for Viscometer before taking viscosity measurement.
5.3.9	The auto zero shall be performed every time the instrument is turned ON.
5.3.10	The operator must ensure that the viscometer is in level by observing bubble in centre position and should remove any attached spindle before performing auto zero.
5.3.11	Press “Next” option from screen to performed Auto Zero.
5.3.12	After the auto Zero is complete, Press “Next” option from screen to Configure Viscosity Test screen.
5.3.13	After the auto Zero is complete, go for Oscillation Check before performing analysis of sample.
	Note: Do not touch the viscometer during the Auto zero process to ensure the best zero value.
	HOME SCREEN:
5.3.14	The Home screen can be accessed by using the Home Icon
5.3.15	The Home screen shows the Main Menu functions and provides access to User Log In and Settings
5.3.16	Configure Viscosity Test	Create and Run viscosity tests.
	Load Test	Load a test that has previously been saved or created with PG Flash software. Tests may be loaded from internal memory or a USB Flash Drive.
	View Results	Load Results (saved test data) that have previously been saved. Results may be loaded from internal memory or a USB Flash Drive.
	Manage Files	Manage the file system in the internal memory or on a USB Flash Drive for test programs and saved data. Create new folder structures, delete files, rename files and move files.
	External Mode	Direct the instrument to communicate with Brookfield Rheocalc T Software for complete viscometer control through software.
	Configure Viscosity Test :

 

5.3.17	The Command Keys include Clear, Save and Run. CLEAR-   Clear all data that has been entered into the test   parameters and restore the values to the factory default. SAVE-   Save the current test. RUN-   Run the current test.
	RANGE :
5.3.18	The Range is also shown in the Running Viscosity Test  during the measurement. Viscosity will be displayed in the unit of measure specified in Settings and is set to centipoise (cP).
5.3.19	RANGE
5.3.20	The Viscometer will calculate the measurement range for a specific Spindle and speed combination. This information is displayed on the screen while selecting the spindle number.
	OUT OF RANGE :
	Note: When the %Torque reading exceeds 100% (over range), the display of %Torque, Viscosity, and Shear Stress will be EEEE
5.3.21	If the %Torque value is between 0 – 9.9%, the data field label will flash.
5.3.22	When the %Torque is below zero(negative values), the display of Viscosity and Shear Stress will be —-.
	Note: Instrument recommends collecting data only when the % Torque reading is between 10 – 100%.
5.3.23	The Viscometer will give on screen indications when the measurement is out of range of the instrument  and  the instrument will display as:
	Note: Measurement data should not be collected when the %Torque reading is out of range. The out of range condition can be resolved by either changing the speed (reduce speed when reading is out of range: high) or changing the spindle (increase the spindle size when the reading is out of range: low).
	OPERATION FOR MAKING MEASUREMENT
	1 )Viscometer
5.3.24	The instrument should be turned ON, leveled.
5.3.25	The level is adjusted using the two feet on the bottom of the base and confirmed using the bubble level on the front of the head of instrument.
5.3.26	Adjust the feet until the bubble is inside the center target. Set the bubble level priorto auto zero and check the level prior to each measurement.
	Note: The proper level is essential for correct operation of the Viscometer.
	2) Sample
5.3.27	The fluid to be measured (sample), must be in a container. The standard spindles supplied are designed to be used with 600 ml beaker or equivalent container with an internal diameter of 8.25 cm/height 12.1cm OR larger container may be used.
	3)Selecting a Spindle/Speed
5.3.28	If the spindle number and speed is already mentioned for the sample, then put directly in test method and carry out test.
5.3.29	If not, then the process of selecting a spindle and speed for an unknown fluid is normally trial and error.
	4) Spindle Entry code and Range Coefficients
5.3.30	The range coefficient is a convenient tool for quickly determining the maximum viscosity that can be measured with a specific spindle/speed combination.
5.3.31	Divide the Range Coefficient by the spindle speed to determine the maximum viscosity in centipoise that can be measured.
5.3.32	Spindle Entry Code and Range Coefficients:- ( as per operational manual Table ) Spindle Entry Code Range Coefficient LV1 61 6,000 LV2 62 30,000 LV3 63 120,000 LV4 64 600,000
5.3.33	E.g. LV Viscometer with LV2 spindle: Range Coefficient is 30,000. At 60 RPM, the maximum viscosity that can be measured is 30,000/60 or 500 cP.
5.3.34	Two general rules are there in the trial and error process. 1)    Viscosity range is inversely proportional to the size of the spindle. 2)    Viscosity range is inversely proportional to the rotational speed.
5.3.35	Viscometers are provided with a set of four spindles and a narrow guard leg
5.3.36	The spindles are attached to the viscometer by screwing them onto the coupling nut on the lower shaft.
5.3.37	Note that the spindles have a left-hand thread. The lower shaft should be secured and slightly lifted with one hand while screwing the spindle to the left. The face of the spindle nut and the matching surface on the lower shaft should be smooth and clean to prevent eccentric rotation of the spindle.
	Note: To avoid the trapping of air bubble beneath the disc, First immerse the spindle at an angle, and then connect it to the viscometer without taking out spindle from the sample solution.
5.3.38	Spindles can be identified by the number on the side of the spindle coupling nut as shown in picture:
	Note: Spindle should dip up to the mark to get accurate reading.
5.3.39	The repeatability viscosity should control the following parameters: 1)      Test temperature  2) Sample container size  3) Viscometer model 4) Sample volume 5)Spindle used  6)Whether or not to attach the guard leg 7) Test speed or speeds 8) Length of time or number of spindle revolutions to record viscosity 9) How the sample was prepared and/or loaded into the container.
	5)Multiple  Data Points
5.3.40	The majority of viscosity and yield stress measurements are made at the quality control level consist of a single data point. Many fluids exhibit a characteristic change in viscosity and stress with a change in applied force.
5.3.41	Non-Newtonian flow is analyzed through the collection of viscosity data over a range of shear rates and the generation of a graph of viscosity versus shear rate.
5.3.42	For collecting multiple data points for comprehensive analysis of flow behavior when using test programs created with PG Flash Software.
	6) Selecting Data Collection :
5.3.43	The Data Collection setting is shown in Configure Viscosity Test directly under the Temperature display. Pressing the blue arrow, in this field, will present the Data Collection screen :
	a ) Single point :
5.3.44	Collect only a single data point when the End Condition is met.
	b) Single Point Averaging:
5.3.45	Specify an amount of time over which to average measured data. Collect a single data point when the End Condition is met.
5.3.46	Example 1: The End Condition is Time with a value of 1 minute and 30 seconds, the Single Point Averaging Duration is 30 seconds, the single data point collected from this step will be an average of the data measured from 1 minute to 1 minute 30 seconds.
	c)Multi Point:
5.3.47	Collect multiple data points based on time. The Data Interval is specified in Hours:Mins:Secs. If the End Condition is set to Time, then the total number of points will be calculated and displayed in the Data Collection screen.
5.3.48	Example 1: End Condition is Time = 2 minutes, Multi Point Data Interval is 10 seconds. Total points collected will be 12 with the last data point taken in the last second of the step.
5.3.49	Example 2: End Condition is set to Viscosity = 200 cP, Multi Point Data Interval is 10 seconds. During the test the total time required to reach 200 cP is 65 seconds. Total points collected will be 6 with the last data point taken at 60 seconds, 5 seconds before the test is finished.
	d)Multi point Averaging :
5.3.50	Specify an amount of time over which to average measured data. Collect multiple data points based on time until the End Condition is met.
5.3.51	Example 1: End Condition is Time = 2 minutes, Multi Point Data Interval is 10 seconds. Averaging Duration is 5 seconds. Total points collected will be 12 with the last data point taken in the last second of the step. Each data point will be an average of the data measured in the last 5 seconds of each Data Interval.
	7) End Condition :
5.3.52	The completion of a test is defined by the End Condition. Each time that you enter Configure Viscosity Test, the End Condition will be set to the last value used. The End Condition parameter or values can be changed by pressing the End Condition button.
5.3.53	The End Condition can be changed by pressing the blue down arrow within the parameter field.
5.3.54	Six End Conditions are available: 1)Time 2)# of Points 3)# of Revolutions 4)Torque 5)Viscosity 6)Temperature
	1)Time:
5.3.55	The test will complete when the specified amount of time has elapsed. Time is entered in Hours, Minutes, and Seconds. refer point 5.3.54
	2)# of Points :
5.3.56	The test will complete when the specified number of data points has been collected. Data is collected according to the Data Collection setting refer point 5.3.57. The range of data points is: 1 – 9,999.
	3)# of Revolutions:
5.3.57	The test will complete when the specified number of revolutions of the spindle has occurred. Data is collected according to the Data Collection setting refer point 5.3.57. The range for number of revolutions is 1 – 9,999.
	4)Torque:
5.3.58	The test will complete when the specified Torque value is measured. Data is collected according to the Data Collection setting refer point 5.3.54. The range of values for the measured Torque is: -10.0 % – 100.0 %
	5)Viscosity:
5.3.59	The test will complete when the specified Viscosity value is measured. Data is collected according to the Data Collection setting refer point 5.3.54. range of measured Viscosity is 0 – 10,000,000,000 cP.
	6)Temperature:
5.3.60	The test will complete when the specified Temperature value is measured to within the indicated Tolerance. Temperature is measured through connected temperature probe.
	Note: A small tolerance value will require a much longer time to reach the End Condition.
	Note: Audible alarms may be turned off in the User Settings
	9) Running a Test:
5.3.61	A viscosity test is started by pressing the Run button on the Configure Viscosity Test screen.
5.3.62	When Run is pressed, the display will change to the Running Viscosity Test screen and the display shows:
5.3.63	The Running Viscosity Test screen provides information on the current measurement Consist : 1)Torque                2)Viscosity                3)Shear Stress 4)Shear Rate            5)Temperature                6)Speed
	1) Torque:
5.3.64	It is described as a percent (%) and has a range of 0 – 100%. The instrument provide measurement results within the stated accuracy provided the Torque reading is between 10 and 100%. If the Torque reading falls below 10% the labels in the data fields will flash to indicate an error condition.
	2)Viscosity :
5.3.65	Viscosity is calculated from the measured Torque based on the selected spindle and speed of rotation.
	3)Shear Stress:
5.3.66	Shear Stress is calculated from the measured Torque based on the selected spindle.
	4)Shear Rate:
5.3.67	Shear Rate is calculated from selected speed based on the selected spindle.
	5) Temperature
5.3.68	Temperature is the input value from a connected temperature probe. The provided probe shall be inserted into the test sample to check the temperature.
	6) Speed
5.3.69	Speed is the selected speed from the Configure Viscosity Test screen.
5.3.70	The More/Less Bar can be used to reduce the number of parameters shown in the display.
5.3.71	The Speed parameter is active while running a test. The operator can change the speed of test without returning to the Configure Viscosity Test screen. refer point 5.3.71
	Note: If the speed is changed during the execution of a saved test, then the test status will be changed to “Unsaved Test”.
	7) Results:
	Return to the Home Menu.

5.3.72	Each data point includes: Viscosity, Torque, Speed, Temperature, Time, Shear Stress (SS), Shear Rate (SR), Density, and Accuracy.
	c)Auto Zero:
5.3.73	Auto Zero is an operation performed by the Viscometer automatically during the power up sequence. This operation sets to a correct zero value.
5.3.74	Selecting Oscillation Check will present a screen instructing to level the viscometer and remove the spindle. After Confirmation of level will present the Oscillation Check screen as shown below.
5.3.75	A successful Oscillation Check will show a very smooth decrease of %Torque with a final value within +/-0.2 of 0.0. Any value above 0.2 % or below -0.2% indicates that a calibration check should be performed
	a)Measurement Units:
5.3.76	The Measurement Units menu displays current selections for measurement units. Each unit can be changed by pressing the down arrow and selecting the desired value.
5.3.77	Measurement Units are part of the data file for saved results. Available units include: 1)Viscosity 2) Speed 3) Temperature 4) Stress (Shear) 5) Density
5.3.78	1)Viscosity
	cP		Centipoises
	P		Poise
	mPa•s		milliPascal seconds
	Pa•s		Pascal seconds
	cSt		Centistokes
	mm2/s		millimeters squared per second
	Note: When cSt units are chosen for viscosity, a density value must be entered in the Density field in Configure Viscosity Test.
	2) Speed
5.3.79	RPM			revolutions per minute
	1/s			reciprocal seconds
	3) Temperature
5.3.80	C				Centigrade
	F				Fahrenheit
	4) Stress (Shear)
5.3.81	Dyne/cm2	Dyne per square centimeter
	N/m2	Newton / square meter
	Pa	Pascal
	5) Density:
5.3.82	g/cm3	gram / cubic centimeter (1gm/cm3)
	kg/m3	kilogram / cubic meter (1kg/m3)
	Note: Density and viscosity are both sensitive to temperature. When using a density value in Configure Viscosity Test, enter a density value that was determined at the same temperature as that of the viscosity measurement.
	d)Spindle List:
5.3.83	The selection of Spindle in Configure Viscosity Test can be done by using either a number pad or a scroll list.
	e)Speed List:
5.3.84	The selection of Speed in Configure Viscosity Test can be done by using either a number pad or a scroll list.
5.3.85	When scroll list is selected, the speed entry is restricted to the speed sets for the LV viscometers. The contents of the scroll list are defined in the Speed List menu.
5.3.86	The  Brookfield speed sets are defined below
	LV					0.3, 0.6, 1.5, 3, 6, 12, 30, 60

 

	2)Users and Access
5.3.87	The instrument can be set up with User accounts to restrict access and enhance data tracking. Two user levels are available in the instrument and each can be customized by the Administrator.
5.3.88	All user accounts require a password for access to the instrument.
5.3.89	The User Settings are divided into three categories; General Settings, Data Access Functions, and Viscosity Test Setup.
5.3.90	The administrator can customize both the Power User and User through the Power User Level Access and User Level Access in the Users and Access Menu.
5.4	Calibration
	Frequency: Quarterly calibration
5.4.1	Generate the A. R. No. on the “Instrument/Equipment Calibration Register” as per SOP and give requisition to QA Department for calibration formats of instruments on the due date of calibration.
5.4.2	For issuance of calibration report refer SOP “Control of documents, Preparation, Approval, Issuance and Maintenance.
5.4.3	Perform the Oscillation Check before Perform the calibration of instrument. Refer point no. 5.3.115
5.4.4	Perform the calibration of instrument as per Annexure and procedure mentioned below.
5.4.5	Calibration of Digital Viscometer is performed for below parameters: 1)Temperature Calibration 2) Spindle Calibration By Using Viscosity Standard Fluid
	1)Temperature :-
5.4.6	The instrument should be turned on, leveled.
5.4.7	The level is adjusted using the two feet on the bottom of the base and confirmed using the bubble level on the front of the head.
5.4.8	Adjust the feet until the bubble is inside the center target. Set the level prior to auto zero and check the level to each measurement and perform auto zero.
5.4.9	Allow 10 minutes warm up time before operation of Digital Viscometer.
5.4.10	Temperature data will only be displayed if a Brookfield temperature probe is connected to the Viscometer.
5.4.11	The Temperature value is measured to within the indicated Tolerance. Temperature is measured through connected temperature probe.
5.4.12	A suitable Viscosity standard fluid is require to perform the calibration performance check.
5.4.13	Brookfield Viscosity Standard Fluids having a validity of one year from date of initial use.
5.4.14	Viscosity Standard Fluids can be stored under normal Laboratory condition.
	Note: 1) For Viscosity Standards < 30,000 cP, use a 600 mL Low Form Griffin Beaker having a working volume of 500 ml. 2) For Viscosity Standards ≥ 30,000 cP, use the fluid container.
5.4.15	Pour the viscosity standard fluid into a 600 ml beaker while being careful to not trap any bubbles.
5.4.16	Place beaker into a constant temperature Bath set to 25°C
5.4.17	Immerse the temperature probe in the liquid for 1 hour
5.4.18	After 1 hour, check the temperature of the viscosity standard fluid with a calibrated thermometer.
5.4.19	Enter the details in calibration report. Refer Annexure No.:- 01
5.4.20	Ensure that the results obtained, comply with the acceptance criteria.
	Acceptance Criteria :- 25.0 ± 0.1°C
	2) Spindle Calibration By Using Viscosity Standard Fluid :-
5.4.21	A suitable Viscosity standard fluid is require to perform the calibration performance check.
	Note: LV #4 (64) spindles have been omitted to perform a calibration check on instrument from procedure. Reasons that the small amount of spindle surface that makes contact with the viscosity standard, the difficulty of establishing the immersion mark precisely and the need for precise temperature control at 25°C in the immediate range of the spindle.
5.4.22	Pour the viscosity standard fluid into a 600 ml beaker while being careful to not trap any bubbles.
5.4.23	Place beaker into a constant temperature Bath set to 25°C
5.4.24	Connect the spindle No. 61 to the viscometer
5.4.25	Lower the viscometer head into position. Be sure to use supplied guard leg.
5.4.26	The viscometer Standard fluid, together with the spindle, should be immersed in the bath for a minimum of 1 hour, gently stirring the fluid.
	Note: spindle should be immersed in standard fluid up to the mark to get accurate reading.
5.4.27	Enter the details of sample in the instrument such as spindle number, Temperature, RPM, end condition etc.
5.4.28	After 1 hour check the temperature. After attaining temperature 25.0°C ±0.1°C. Press the run option from test panel.
	Note: The spindle must rotate at least five times before readings are taken.
5.4.29	For spindle No. 61 measure the viscosity using three rotational speeds. Such as 1 RPM, 2 RPM, 4 RPM.

 

5.4.30	Constant values of SMC and TK:- Spindle Entry Code SMC TK ( For DV2TLV Model) LV1 61 6.4 0.09373 LV2 62 32 LV3 63 128 LV4 64 640 SMC = Spindle Multiplier Constant ; TK = Torque Constant Calculation of Full Scale viscosity Range (FSR) :-         FSR (cP) =        TK * SMC* 10000_                                          RPM
5.4.31	Same procedure to be followed for Spindle no. 63 from point no. 5.4.5 to 5.4.30. For spindle No. 63 .Measure the viscosity using three rotational speeds. Such as 10 RPM, 30 RPM, 60 RPM.
	Interpretation of Calibration Test Results:
5.4.32	The viscosity reading should equal the cP value on the fluid standard to within the combined accuracy of the viscometer and the viscosity standard.
5.4.33	Brookfield Viscosity Standards Fluids are accurate to (+/-) 1% of their stated value. The instrument is accurate to (+/-) 1% of the range when using spindles LV #1-3.
5.4.34	Example: Calculate the acceptable range of viscosity using DV2TLV with LV-3 Spindle at 20 RPM Viscosity Standard Fluid 1000 with a viscosity of 1005 cP at 25°C: 1) Calculate full scale viscosity range using the equation: Full Scale Viscosity Range [cP] = TK * SMC * 10,000                       RPM Where: TK – 0.09373 from point no 5.4.30 SMC =128 from point no. 5.4.30   Full Scale Viscosity Range = 0.09373* 128 * 10,000 = 6,000 cP                  20 The viscosity is accurate to (+/-) 60 cP (which is 1% of 6,000) 2) The viscosity standard fluid is 1005 cP. Its accuracy is (+/-)1% of 1005 or(+/-)10.05 cP. 3)Total allowable error is (10.05 + 60) cP = (+/-) 70.05 cP. 4)Viscosity reading between 935 and 1075.1 cP indicates that the Viscometer is operating correctly. Reading outside these limits may indicate a Viscometer problem.
5.4.35	For more information of Constant values of SMC and TK refer operational manual Table D-1 and Table D-2 at page 68,69,70
5.4.36	Enter the details in calibration report. Ensure that the results obtained, should complies with the acceptance criteria. If instrument fails to meet to acceptance criteria then file an incident and investigate the cause of failure.
5.4.37	If calibration activity is outsourced, the values are to be transcribed in the Annexure No. 01, and external party certificate to be attached.
5.5	Cleaning Procedure:
5.5.1	Make sure that the instrument is in a clean, dry working environment (dust-free, moderate temperature, low humidity, etc.).
5.5.2	Make sure the instrument is on a level surface.
5.5.3	Hands/fingers must be clean and free of sample residue. Not doing so may result in deposit build up on the upper part of the shaft and cause interference between the shaft and the pivot cup.
5.5.4	Be sure to remove the spindle from the instrument prior to cleaning. If the spindle is cleaned in place Severe instrument damage may occur.
5.5.5	Instrument and display Clean with a dry, non-abrasive cloth. Do not use solvents or cleaners. The instrument housing is manufactured from polycarbonate ABS. Clean instrument housing with mild soap and water.
5.5.6	Spindles are made of stainless steel. Clean with a non-abrasive cloth and solvent appropriate for sample material.
5.5.7	When cleaning, do not apply excessive force, which may result in bending spindles.

 

6. Definitions / Abbreviations:

• Definitions:
• Calibration: Tests performed to ensure that the instrument is functioning as per set parameters. These tests are performed as per predefined frequency and also after any major repair or replacement of parts.
• Viscosity: – A quantity expressing the magnitude of internal friction in a fluid, as measured by the force per unit area resisting uniform flow.

 

• Digital Viscometer: – Viscosity is the measure of the internal friction of a fluid. This friction becomes apparent when a layer of a fluid is made to move in relation to another layer. The greater the friction, the greater the amount of force required to cause this movement, which is called shear.

 

• Abbreviations:

Abbreviation	Expansion
±	Plus or Minus
No.	Number
A.R.No.	Analytical Reference Number
mg	milligram
ml	millilitre
cm	Centimetre
cP	Centipoise
CFR	Code of federal regulation
vs	verses
°	Degree
FSR	Full Scale Range
SRC	Shear Rate Constant
SMC	Spindle Multiplier Constant
RPM	Rotation per minute

 

Calibration Report

1. TEMPERATURE CALIBRATION:-

Description of test	Set Temperature	Acceptance Criteria	Observation		Remarks
			Internal Temperature Probe	External Temperature Probe
Temperature	25.0 ºC	25.0 ± 0.1 ºC

 

2. SPINDLE CALIBRATION BY USING VISCOSITY STANDARD FLUID:-

 

1. Standard Fluid Viscosity as per Certificate = ________cP (A)

 

2. 1 % value of Standard Fluid Viscosity =         A * 1___ 

                                                                              100

                                                                      

                                                           =____________________ (B)

3. Calculation of Full Scale viscosity Range (FSR) :-

 

                                             FSR (cP) =        TK * SMC* 10000_   

                                                                                RPM

 

*Note: – For value of Torque Constant (TK) and Spindle Multiplier Constant (SMC) refer SOP  

 

1. Calculation of Viscosity Acceptable Range :-

 

Low Limit	High Limit
A – (B + C)	A + (B + C)

 

1. Viscosity check for Spindle  :-

Spindle Number	RPM	Observed Viscosity (cP)	Observed    Torque (%)	FSR	1% of FSR (C)	Acceptance Criteria		Remark
						Low  Limit	High Limit
61	1
	2
	4

 

2. Viscosity check for Spindle :-

Spindle Number	RPM	Observed Viscosity (cP)	Observed    Torque (%)	FSR	1% of FSR (C)	Acceptance Criteria		Remark
						Low  Limit	High Limit
63	10
	30
	60

 

Remark: The result Complies/Does not complies as per acceptance criteria.

 

Performed By: ________________         Date: ____________         Checked By: ______________                Date: ______________

                                                     

 

SOP Title: Operation, Cleaning and Calibration of Automatic  Polarimeter

 

1. Objective:

To lay down the procedure for Operation, Cleaning and Calibration of Automatic Polarimeter.

 

2. Scope:

This SOP is applicable for Operation, Cleaning and Calibration of Automatic Polarimeter located in Quality Control Laboratory.

 

3. Responsibility:

• Quality Control: To prepare and review the SOP. To follow the procedures laid down for Operation, Cleaning and Calibration of Polarimeter as per this SOP.
• Engineering Department: To carry out preventive maintenance as per schedule and procedure defined.
• Quality Assurance Department: To review and approve the SOP and Annexure.

 

4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

 

5. Procedure :

 

5.1	Safety Precautions/Instructions:
5.1.1	Ensure all the electrical connections are properly earthed. Ensure the power plug of the automatic Polarimeter is fixed in the socket properly.
5.1.2	Wear personal protective equipments such as safety glasses, hand gloves during operation of instrument.
5.1.3	Ensure that the ON/OFF switch is working properly by switching “ON” the Polarimeter.
5.1.4	Clean the Automatic Polarimeter with dry lint free duster.
5.1.5	Do not move the automatic Polarimeter during cleaning
5.1.6	Do not attempt to do any repairs unless trained.
5.1.7	After switch ON instrument wait for 10 minute to allow stable temperature.
5.1.8	Ensure sample cell foot is clean.
5.1.9	Clean the sample cell and temperature transmission surface with isopropyl alcohol
5.1.10	Fill sample cell with sample solution in such a way that, there should not be any gap/empty space at both ends. Then cap the both ends and observe from lens of Polarimeter cell that no air bubble present inside the cell.
5.1.11	Take sample reading within 30 minutes of sample preparation because some compounds are optically unstable and may impact on the results.
5.2	Operational Procedure:
5.2.1	Switch ON the mains and the Automatic Polarimeter switch.
5.2.2	Press “Zero” key to zero the instrument which is present on main screen after the instrument is just ON.
5.2.3	Ensure the instrument is clean and free from dust.
5.2.4	Check due date of instrument calibration before starting measurements.
5.2.5	Wait for 10 minute to allow stable temperature.
	Note: For Sample measurement two paths are given 1. By selecting main window screen parameter like Temperature, Scale, Wavelength, measurement mode in setup menu.
	2. By Selecting Sample Method
	For Sample Measurement
5.2.6	Click on ‘Scale’ key on main screen. It contains measurement  option 1. Optical rotation       2. Specific optical rotation                           3. Concentration 4. Specific rotation plus         5. ‘Z’ Scale (589)
5.2.7	If scale ‘optical rotation’ selected then a display  shows parameters 1. Cell Length   2. Multiplier
5.2.8	Select cell length 100 mm and multiplier as ‘1’& click on ‘Set’ key .
5.2.9	If Scale ‘Specific Optical rotation’ selected then the  display will show Parameter 1. Cell length   2. Concentration
5.2.10	Select cell length 100 mm and put concentration as g/100 ml and click on set key. e.g. 10 % Sucrose solution then concentration value is 10.0
5.2.11	Note: 1.   “Specific Rotation plus” mode is selected only when measuring sample has water content/ Loss on drying. The multiplier used to compensate for changes in a sample due to loss on drying/water content. Put “Multiplier” value by manually calculating by formula as 100/(100-water /LOD) e.g. Paracetamol Water content is 5.5 then multiplier value  is 100/(100-5.5) = 1.06 2.    If scale “Specific Rotation plus” selected then a display will show parameter 1.Cell length 2. Concentration 3. Multiplier 3.    Put  value of parameter and click on “SET” key.
5.2.12	Click on ‘Temperature’ key on main screen display. Then screen  shows parameters as 1. Temperature Control  2.Temperature correction 3.TempTrol                     4.Water bath
5.2.13	Select the ‘Temperature Control’ parameter. Then click on “TempTrol”  option and select input temperature as 20ºC or 25ºC as per sample requirement and click on ‘Set’ key.
	NOTE: Put Temperature control option “ON” only when, in-built temperature probe kept in sample cell with sample solution. Otherwise keep “OFF” when in-built temperature probe is placed in the cap holder.
5.2.14	Select the Measurement mode: Single or Multiple from the Measurement mode menu in the Setup menu present on main screen.
5.2.15	If  single selected- It Measures  sample one time If Multiple mode selected, Enter the no. of measurement selected between 1 and  999 . Several measurement of sample made.  For routine analysis  ‘5’ number of measurement shall be selected.
5.2.16	Press the ‘Wavelength ‘key on main screen and select the desired wavelength.
5.2.17	Fill sample cell with sample blank (Diluent) and keep in sample chamber. Put in-built temperature probe in sample cell   and close the door of sample compartment.
5.2.18	After attaining set temperature click on ‘Zero’ key on main screen to nullify blank interference.
5.2.19	Remove blank solution from sample cell and rinse the cell with sample solution for 2-3 times.
5.2.20	Fill sample cell with measuring sample solution.   Care should be taken that no air bubble will be observed inside wall of the cell.
5.2.21	Keep sample cell with sample solution in sample chamber. Place in-built temperature probe in it.
5.2.22	After attaining required sample temperature on digital display press “Start” key from main screen. The system will prompt for sample ID. Put sample ID and press “Enter”
5.2.23	After completion of measurement, the result will display on main screen.
5.2.24	To save or Print the measurement result click on ‘printer’ key on main screen window. Results will get saved in of ‘C’ drive with auto generated file name.
5.2.25	Remove the sample cell from chamber; flush it with water and isopropyl alcohol. Clean outside surface of sample cell foot and temptrol surface   with isopropyl alcohol.
	Note: 1.   “Specific Rotation plus”  mode is selected only when measuring sample has water content/ Loss on drying. The multiplier used to compensate for changes in a sample due to loss on drying/water content. Put “Multiplier” value  manually calculating by formula as 100/(100-water/LOD)
5.3	Calibration Procedure:
	Frequency: Quarterly or after carrying out any major maintenance work.
5.3.1	Generate the A. R. No. on the “Instrument/Equipment Calibration Register” as per SOP and give requisition to QA department for calibration formats of instruments on the due date of calibration.
5.3.2	For issuance of calibration report refer SOP “Control of documents, Preparation, Approval, Issuance and Maintenance.
5.3.3	Perform the calibration of instrument as per Annexure and procedure mentioned below.
5.3.4	Calibrate the instrument using a solution of previously dried sucrose at 105º C for 1 hour. Measure  the optical rotation in a 100 mm tube at 250 C by using the sucrose concentrations as follow: 1) 10 % sucrose solution: Dissolve 10 gm sucrose to 100 ml with water. 2) 20 % sucrose solution: Dissolve 20 gm sucrose to 100 ml with water. 3) 30 % sucrose solution: Dissolve 30 gm sucrose to 100 ml with water. 4) 40 % sucrose solution: Dissolve 40 gm sucrose to 100 ml with water. 5) 50 % sucrose solution: Dissolve 50 gm sucrose to 100 ml with water.
5.3.5	Follow the same procedure of measurement as above mentioned in point number  5.2.1 to 5.2.25
5.3.6	Enter the details in calibration report. Ensure that the results obtained, comply with the acceptance criteria. If instrument fails to meet acceptance criteria then file a incident and investigate the cause of failure.
	NOTE: 1. Acceptance criterion for calibration of instrument by sucrose is given for 2 dm tube in USP/IP. 2. However, 1 dm tube is used in Polarimeter. Hence, manufacturer recommended the sensitivity of reading in 1 dm tube is exactly half of that 2 dm tube. The acceptance criteria is set according to manufacturer recommendation.
5.3.7	Update the “Instrument/Equipment Usage Log Book”, “Annual Calibration Schedule” and “Quarterly Calibration Schedule” as per SOP “Calibration of Instruments and Equipments”.
5.3.8	Affix the “Calibration Status Label” on the Instrument as per SOP “Calibration of Instruments and Equipments”.
5.3.9	If calibration activity is outsourced, the values are to be transcribed in the Annexure, and external party certificate to be attached.

 

6. Definitions / Abbreviations:

• Definitions:
• Calibration: Tests performed to ensure that the instrument is functioning as per set parameters.    These tests are performed as per predefined frequency and also after any major repair or replacement of parts.
  • Temperature Probe : It measure temperature of sample cell and main body of
  • Optical Rotation : Substance are optically in the sense that they rotate incident plane polarized light so that the transmitted light energes at a measurable angle to the plane of the incident light.
  • Polarimeter Principle :

The optical rotation of a substance is the angle through which the plane of polarization is rotated when polarized light passes through the substance. Substances are described as Dextro-rotatory (+) or Laevo-rotatory (-) according to whether the plane of polarization is rotated clockwise or anticlockwise.

• Abbreviations:

Abbreviation	Expansion
SOP	Standard Operating Procedure
QC	Quality Control
QA	Quality Assurance
0C	Degree Celsius
%	Percentage
dm	decimeter
No.	Number
A. R. No.	Analytical Reference Number

 

Calibration Report

Sr. No.	Parameters	Standards	Acceptance Criteria	Observed Values
1.	Optical Rotation (at 25ºC)	10 % sucrose solution	6.665° (6.590° to 6.700°)
		20 % sucrose solution	13.305° (13.180 °to 13.400°)
		30 % sucrose solution	19.930° (19.770° to 20.100°)
		40 % sucrose solution	26.530° (26.360° to 26.800°)
		50 % sucrose solution	33.115° (32.950° to 33.500°)

 

Remark: The result Complies/Does not complies as per acceptance criteria.

 

Performed By: ________________     Date: ___________                Checked By: ______________          Date: ______________

                                                     

 

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.