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Qualification and Validation

Validation: ( related to Process/ Product/Equipment )

Action of proving, in accordance with the principles of GMP, that any procedure, process, equipment, material, activity or system actually leads to the expected results. or

It is a process using documented evidence that provides a high degree of assurance that a specific process will consistently produce the predetermined outcome.

Qualification and Validation

Qualification: (Validation usually incorporates the concept of qualification which is related to the equipment) Action of proving that any premises, systems and items of equipment work correctly and actually lead to the expected results (means, it is used to determine whether the equipment operates as it was designed to in a reproducible manner). or Process of establishing confidence that the equipment is capable of consistently operating within established limits and tolerances. Studies therefore done prior to use

DEFINITIONS-Validation
Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications and quality attributes.

VALIDATION VERSUS VERIFICATION, TESTING, CALIBRATION, AND QUALIFICATION

Qualification
The term qualification has been defined for the installation, operation and running of a system under workload for a specific application. Like verification, qualification is also part of validation and is product-specific.

Verification
Confirmation by examination and provision of evidence that specified requirements have been met. Performance verification of analytical instrumentation is the process of comparing the test results with the specification. It includes testing and requires the availability of clear specifications and acceptance criteria. The verification process ends with the generation and sign-off of a Declaration of Conformity of the instrument to specifications.

Equipment Qualification
Equipment qualification has been broken down into four parts:

Equipment Qualification
DQ, for setting functional and performance specifications (operational specifications); IQ, for performing and documenting the installation in the selected user environment; OQ, for testing the equipment in the selected user environment to ensure that it meets the previously defined functional and performance specifications; and PQ, for testing that the system performs as intended for the selected application.

Qualification timeline

Validation Components
Validation Master Plan (VMP) Project Validation Plan (PVP) User Requirements Specification (URS) Functional Specifications (FS) Design Specification (DS) Design Qualification (DQ) Test Protocols Installation Qualification (IQ) Operation Qualification (OQ) Performance Qualification (PQ) Summary Report(s)

VMP in the V-Model

Validation Master Plan (VMP) Defines

Overarching procedural document Illustrates Company commitment to quality Comprehensive overview of Facilities, Equipment, Processes, Automation, Procedures, & Methods Guide to how validation is performed one approach Defines project responsibilities Defines Validation goals Description of all equipment, systems, procedures, etc. that fall within scope of validation Describes approach company will take to identify all matters that require validation oversight

VMP Components
Introduction

Purpose what is this document? Scope what is included and excluded from governance by the content this document?

Definitions & Abbreviations Approach

Rationale is there standard or model upon which this plan is based (e.g. V-Model, ISO-9001, or other internal policies) Overview a good place to have flowcharts of the validation process For each major process element, describe:

Input(s) into the process Output(s) from the process (e.g. documents) Decision point(s) and possible outcomes Iterative cycle(s) (e.g. document review/approval)

VMP Components
Validation Documentation Lifecycle

Address the way in which documentation generated under this VMP is to be generated and handled

Document Numbering Convention(s) Document Contents & Formats (Templates?) Document Review, Approval, & Tracking Document Archival & Disbursement VMP Maintenance/Review/Archival

Related Guidelines, Policies, & Procedures

What other documents fall under the umbrella of this master plan?

General Topics Existing SOPs Planned / In-Progress SOPs Work Instructions Training Programs Equipment / System / Product Lists

VMP Components
Related Roles and Responsibilities
Try to assign by role, not by name (personnel and job titles change). Common roles include:

Author System Owner Representative Technical Reviewer Subject Matter Expert (SME) Quality Assurance

References

What documents were used to develop this VMP Who needs to review and/or approve this VMP

Distribution

This format can be used for all validation documents

PVP in the V-Model

Validation/Qualification Study Phases

Qualification should be done in accordance with predetermined and approved qualification protocols/plan. The results of the qualification should be recorded and reflected in qualification reports. The extent of the qualification should be based on the criticality of a system or equipment, e.g. Blenders, autoclaves or computerized systems.

Validation Study Phases

Phase 1 : Pre-Validation Phase (Qualification Phase)(DQ&IQ) ; Phase 2: Process Validation Phase (Process Qualification phase)(OQ&PQ); Phase 3: Validation Maintenance Phase. Phase 1 : This phase covers all activities relating to product research and development, formulation, pilot batch studies, scale-up studies, transfer of technology to commercial scale batches, establishing stability conditions, storage and handling of in-process and finished dosage forms, master Equipmentproduction

Qualification(Design&

Installation),

documents, Process Capability.

Phase-1 study (Pre-validation)

In this, Design& Installation of equipment is validated. DESIGN QUALIFICATION: DQ is a documented review of the design, at an appropriate stage or stages in the project, for conformance to operational and regulatory expectations. Specification of requirements for facilities, plants and equipment, mainly in connection with purchase/modification of equipment. INSTALLATION QUALIFICATION( IQ): It verifies that the equipment has been installed in accordance with the manufacturers recommendation in a proper manner and that all of the devices are placed in an environment suitable for their intended purpose. COMPUTER AND SOFTWARE IQ: Software Requirements and Specifications should be clear, written down, and approved by the appropriate personnel prior to creating the software package. Software Installation should be recorded, when made, so that the proper model numbers, serial numbers ,installation date etc.

Design Qualification(DQ)/Review
Design qualification (DQ) defines the functional and operational specifications of the instrument and details for the conscious decisions in the selection of the supplier". Means, it is a documented review of design. Performing a DQ is not a legal requirement, but it should be a requirement of a proper engineering process. Objective: To check the design documents of a technical system to ensure that they fulfill the user requirements.

Points to be considered for inclusion in a DQ

Description of the analysis problem. Description of the intended use of the equipment Description of the intended environment. Preliminary selection of the functional and performance specifications (technical, environmental, safety). User requirements should be considered when deciding on the specific design of a system or equipment. A suitable supplier should be selected for the appropriate system or equipment (approved vendor).

Design qualification-Why ?
Equipment layout and design must aim: to minimize risks of error. to permit effective cleaning and maintenance. To avoid: cross-contamination, dust and dirt build-up any adverse effect on the quality of products Appropriate design easily and thoroughly cleaned on a scheduled basis procedures and records No hazard to the products contact parts of suitable non-reactive materials non additive and not absorptive
Defective equipment removed, or labelled to prevent use

Documentation of DQ
DQ Review Documents: -User Requirement Specifications(URS)
-Functional Specifications(FS) -Tender Specifications& Drawings -Vender Quality Procedures -Purchase specifications The good documentation is the basis for defining tests in the IQ, OQ and PQ Phases.

User Requirement Specification

Objective: To document the needs of the mfg. department & to generate a document detailing all the GMP requirements the technical system has to fulfill. It should written for a technical system that should be implemented in the production process of a pharm. product. A well prepared URS is the key to a projects success. It also provides the basis for an offer to the supplier. Without a comprehensive URS, a pharm. Company cannot get a clear understanding of the supplier and may be led to make a wrong decision. Past experiences such as project faults, inefficient technical systems, and bad commissioning can be included in a URS.

Content of URS
Cover sheet: Reference with numbering system, date and signature of author, approvals with sign and date of user, engineering and QA. Scope: General intention of URS and introduction of focused system. System description: includes brief description of system; description of system boundaries; naming the product produced; outline of technical aspects. Positioning requirements: what requirements the system has concerning location and interfaces to other systems Definition of system specification: description of all important and critical specifications of the system, like process parameters, technical parameters, safety requirements, environmental requirements and GMP relevant specifications Regulatory documents: Reference all relevant regulatory documents that are important for implementing the system.

Installation Qualification(IQ)
Installation qualification establishes that the instrument is received as designed and specified, that it is properly installed in the selected environment, and that this environment is suitable for the operation and use of the instrument. IQ is the method of establishing with confidence that all major processing, packaging equipment and ancillary systems are in conformance with installation specifications, equipment manuals, schematics and engineering drawings. It verifies that the correct equipment has been received and installed as per plan and protocol. Also that it is complete and undamaged (parts, services, controls, gauges and other components.

IQ continue
IQ aims to check documentation against reality and to ensure that the GMP requirements are fulfilled. The generally accepted way to perform an IQ is to, -Develop an IQ protocol -Approve the IQ protocols (by the quality assurance, production and technical dept.) -Perform the IQ -Work out the IQ report -Approve the IQ report (by the quality assurance, production and technical dept.)

IQ includes
Installation qualification requires ,
- A formal and systematic check of all installed equipment against the equipment suppliers specifications and additional criteria identified by the user as part of the purchase specifications. These checks, tests and challenges should be repeated a significant number of times to assure reliable and meaningful results..

IQ includes..
Examination of ,
-equipment design, -determination of calibration- Calibrate the measuring, control and
indicating devices against appropriate, traceable national or international standards

- maintenance and adjustment requirements.

For complicated or large pieces of equipment,

- a pharmaceutical manufacturer may elect to undertake a pre delivery check of the equipment at the suppliers assembly facility. All equipments, gauges and services should be adequately identified and should be given a serial number or other reference number. This number should appear in the reports for the equipment validation studies .

Installation Checklist
Prior to installation: verify physical requirements have been met,

Safety checks, electrical, space, ventilation, water supply, ambient temperature, etc.
confirm responsibility for installation. Upon receipt: verify package contents. do not attempt to use prior to proper installation. If required, ensure the equipment is installed by the manufacturer.

 After installation, Establish inventory record. Define conditions. Develop and implement protocols for calibration, performance verification, and operating procedures. Establish maintenance program. Provide training for all operators.

Documentation of IQ
Indicate satisfactory installation. Documented records for the installation, Installation qualification report Calibration reports Training reports Include details, e.g. The supplier and manufacturer System or equipment name, model and serial number Date of installation Spare parts, relevant procedures and certificate

Phase-2 (process qualification)

In this, operation& performance of equipment is validated It is designed to verify that all established limits of the Critical Process Parameters are valid and that satisfactory products can be produced even under the worst case conditions. OPERATIONAL QUALIFICATION(OQ): It is a documented action of demonstrating process that equipment and ancillary systems work correctly and operate consistently in accordance with established specifications. PERFOMANCE QUALIFICATION (PQ): It Assure that the process continues to comply with established requirements. Performance Qualification (PQ) for testing that the system consistently performs as intended for the selected application.

Operational Qualification (OQ)

This step proceeds after the IQ has been performed. In the OQ, tests are performed on the critical parameters of the system/process. These are usually the independent and/or manipulated variables associated with the system/equipment. All tests data and measurements must be documented in order to set a baseline for the system/equipment.

 Operational qualification (OQ) is the process of demonstrating that an instrument will function according to its operational specification in the selected environment ."

 Operational Qualification (OQ ): The conduct of an Operational Qualification should follow an authorized protocol. The critical operating parameters for the equipment and systems should be identified at the O.Q. stage. The plans for the O.Q should identify the studies to be undertaken on the critical variables, the sequence of those studies and the measuring equipment to be used and the acceptance criteria to be met. Studies on the critical variables should include a condition or a set of conditions encompassing upper and lower processing and operating limits referred to as worst-case conditions. The completion of a successful O.Q. should allow the finalization of operating procedures and operator instructions documentation for the equipment. This information should be used as the basis for training of operators in the requirements for satisfactory operation of the equipment. The completion of satisfactory I.Q. and O.Q. exercises should permit a formal release of the equipment .

Validation
Operational qualification
Systems and equipment should operate correctly operation verified as in the qualification protocol Verifies that the equipment operates consistently within established limits and tolerances over the defined operating ranges. Studies on critical variable to include conditions encompassing upper and lower operating limits and circumstances (i.e. worst case conditions) Challenges equipment functionally to verify compliance with manufacturers specifications and end user defined requirements. To include verification of operation of all system elements, parts, services, controls, gauges and other components

Validation
Operational qualification (2)
Documented records (Operational qualification report) Finalize and approve SOP (operation) Training of operators provided training records Systems and equipment released for routine use after completion of operational qualification, provided that:

All calibration, cleaning, maintenance, training and related tests and results were found to be acceptable

How may runs are to be performed during Operational Qualification (OQ) testing?

Guidelines stress the importance of equipment qualification simulating actual production conditions, including 'worst case' situations and that "tests and challenges should be repeated a sufficient number of times to assure reliable and meaningful results." "three consecutive batches" is recommended for process validation rather than for equipment qualification. No specific number of "runs" for equipment qualification, but multiple tests to simulate actual operating ranges and to establish consistency are expected

Performance Qualification
Performance Qualification (PQ) is the process of demonstrating that an instrument consistently performs according to a specification appropriate for its routine use ".

Performance Validation

Validate the performance of new equipment prior to use, Test known samples, analyze data. Establish stability/uniformity temperature controlled equipment. Check accuracy/precision for pipettors. Check centrifuge rpm.

Validation
Performance qualification
Verifies that the equipment performs according to design specifications and user defined requirements in a reliable and reproducible manner under normal production conditions. Verified in accordance with a performance qualification protocol Documented records performance qualification report Show satisfactory performance over a period of time i,e. carried out long enough to prove that the equipment is under control and turns out product of specified quality consistently. Manufacturers to justify the selected period

Phase-3 (Maintenance Phase)

It is the maintenance of the validation which has been already established. It requiring frequent review of all process related documents, including validation audit reports to assure that there have been no changes, deviations, failures, modification to the production process, and that all SOPs have been followed, including Change Control procedures.

Benefits of a Maintenance Program

Safety Fewer interruptions of work Lower repair costs Elimination of premature replacement Less standby equipment Identification of high maintenance cost Reduction of variation in test results Greater confidence in the reliability of results

Maintenance Program
Systematic and routine cleaning, adjustment, or replacement of instrument and equipment parts, Performed periodically, daily, weekly, monthly. Examples: Cleaning optical lenses Changing motor brushes Thermostat adjustments

Function Checks
Monitoring of instrument parameters to verify that that your equipment is working according to the manufacturers specification. Performed periodically- daily, weekly, monthly. Performed after major instrument repair. Example:

Daily monitoring of temperatures Checking wavelength calibration Checking autoclave indicator paper

Implementing a Maintenance Program

Assign responsibility, Oversight of all laboratory equipment. Individual responsibilities. Develop written policies and procedures. Train staff. Keep records.

Implementing a Maintenance Program

Create a record for equipment inventory, Name, Model #, Serial # Location in lab Date purchased Manufacturer and vendor contact information Warranty, expiration date Spare parts For each piece of equipment,

Establish routine maintenance plan Establish required function checks Develop a list of spare parts

Implementing a Maintenance Program: Spare Parts

Establish and maintain an inventory of most frequently used spare parts. Include in record of inventory:

Spare parts per equipment Part number Average use Minimal # of items to be stored Cost and date of ordering Dates of entry and issuance of part from inventory stock Balance of items remaining in inventory

Implementing a Maintenance Program: Documents

Develop written procedures for all equipments, Concise step-by-step instructions for performing maintenance and function checks Include guide for troubleshooting Establish maintenance records to track: function checks and routine maintenance calibration manufacturers service

Re-Validation and Retro-Validation

Re-Validation: It is required when the operating equipment or system has been changed in some way. It is carried out when major mechanical equipment has been replaced/ Computer systems have been replaced/ New products have to be run on the system/ Critical items have been replaced or repaired. Retro-Validation( Retroactive validation): It must be done on equipment that records have either been lost or validation was not provided for in the initial life of the machine. Retroactive validations get more difficult if the older the machine.

Equipment
Possible Issues
Poor design Lack of safety Poor quality finishes Lack of cleaning Lack of maintenance No usage log or record Use of inappropriate weighing equipment Open-plan location of compressing machines

Project Validation Plan - PVP

PVP is a:
Structured way to evaluate and communicate the necessary validation elements of a complex project or system Useful means for driving the project schedule Method for obtaining pre-approval for a lot of work through stakeholders before investing a lot of time. Meaningful tool that should be used on a value-added basis

VMP vs. PVP

VMP applies to all validation projects PVP applies to a specific project

calibration of various analytical instruments

Calibration
Calibration is one of the primary processes used to maintain instrument accuracy. Calibration is the process of configuring an instrument to provide a result for a sample within an acceptable range.

Calibration is the comparison of a measurement device against a standard.

Calibration can be done:with a new instrument when a specified time period is elapsed when an instrument has had a shock or vibration sudden changes in weather when observations appear questionable

Introduction
A pH meter requires calibrating to give accurate pH readings. A pH meter calculates a sample's pH, based on the Nernst equation A 2 or 3 point calibration, using 2 to 3 different buffer solutions is usually sufficient for initial calibration as the meters electronic logic will calculate the pH values in between. A 1 point calibration, with a buffer of a pH closest to the expected sample pH should be adequate once per day, but a 2 point calibration is recommended.

Procedure
One point
Using pH 4.01

Two point
Using pH 7.0 and pH 4.01

Three point
Using pH 4.01, 7.0 and solution having pH close to required solution

pH chart
DEGREES-Co 0 5 10 15 20 25 30 35 40 45 50 55 60 70 80 90 DEGREES-Fo 32 41 50 59 68 77 86 95 104 113 122 131 140 158 176 194 pH 7.12 7.09 7.06 7.04 7.02 7.00 6.99 6.98 6.97 6.97 6.98 6.98 6.98 6.99 7.00 7.02

Calibration of HPLC:The calibration of HPLC system shall be performed in the first week of every three months.

Calibration of the pump Column compartment thermostat / oven calibration Detector lamp intensity Pump Precision Check Calibration od detector wave length accuracy Check Detector Linearity Calibration of Refractive Index Detector Auto injector linearity

Pump calibration
Check the flow rate of the filtered and degassed Purified water as follows:
Disconnect any column if connected to the system and connect restriction capillary

1.3 Set the flow of the pump at 1.0 mL per minute. Keep the system at this flow rate for about 5 minutes to equilibrate the system. Collect the volume of water delivered for 5 minutes and determine the weight of the water . Convert the weight in to volume by the following table:

Calibration of the pump:-

Temperature, C 20 21 22 23 24 25

Volume, in mL, of 1g of water 1.0028 1.003 1.0032 1.00345 1.0037 1.0039

Temperature, C 26 27 28 29 30

Volume, in mL, of 1g of water 1.0042 1.0045 1.0047 1.005 1.0053

Flow =

Volume Obtained Time measured

Acceptance criteria for Flow rate: + 0.1mL

Column compartment thermostat / oven calibration :Disconnect the column and connect with restriction capillary to the system with flow rate 1.0 ml/min. Keep the sensor wire of the traceable digital thermometer in the right side of the column thermostat/ oven. Set the temperature at 10.0C. Wait for 5 minutes before readings are taken so that the temperature on the Instrument and that displayed on the thermometer are stable. Note down the temperature displayed by the instrument and the thermometer. Repeat the steps for temperature settings at 20C,30.0C, 40.0C,50.0C 60.0C& 70.0C. Shift the thermometer sensor wire to the left side thermostat and repeat the exercise Acceptance Criteria: The difference between the set temperature and the displayed temperature on the thermometer should not be more than 2.0C

Calibration of the Detector Lamp Intensity

Pump Precision Check :-

Chromatographic Conditions:
Column Wave length Injection volume Flow Rate Run time Temperature Mobile Phase : Hypersil C18 100mm X 4.6mm ID : 273 nm : 20 l : 1 ml/min : 5 min : Ambient : Methanol HPLC Grade

Stock Solution Preparation Standard Preparation. Inject 20 uL blank as a Methanol in to the system and record the chromatogram. Program the data processor to inhibit the integration of peaks due to blank Inject 20 l of standard solution (0.1mg/ml) for six times in to the system and record the chromatogram. Calculate the %RSD for Retention Times and Areas observed from six replicate injections. Acceptance Criteria: % RSD calculated for Retention times and areas should not be more than 1.0%

CALIBRATION OF DETECTOR
Wave length accuracy Check :-

Set up the chromatographic conditions apart from wavelength parameter as mentioned in the pump precision check. Inject 20 l of methanol as a blank into the system and record the chromatogram. Inject 20 l of standard solution at 268 nm and record the chromatogram. Similarly inject 20 l of standard solution by increasing 1 nm wavelength upto 278 nm and record the chromatograms. Determine the absorbance level at every nanometer.

Acceptance Criteria: The maximum absorbance level should be at 273 + 1 nm

only.

Detector Linearity:Prepare the conc.0.02mg/ml, 0.04mg/ml, 0.06mg/ml, 0.08mg/ml, 0.1mg/ml. solutions from the stock solution. Inject 20 ml methanol as a blank and record the chromatogram. Inject 0.02mg/ml, 0.04mg/ml, 0.06mg/ml 0.08mg/ml, 0.1mg/ml concentration solutions into the system and record the chromatograms. Take the areas for the concentrations of 0.02mg/ml, 0.04mg/ml, 0.06mg/ml 0.08mg/ml, 0.1mg/ml and plot a graph for concentration (mg/ml) Vs area counts.

Acceptance Criteria: Correlation coefficient should be 0.99 to 1.00

Calibration of Refractive Index Detector :Chromatographic Conditions: Column : Hypersil 100mm X 4.6 mm ID Flow Rate : 1.5 ml/min Runtime : 8 min Oven temperature : 40oC Mobile phase : A filtered and degassed mixture of acetonirile and water in the ratio of 8:2 Preparation of Standard (200mg anthracene in 100 ml water) Equilibrate the column for 30 minutes with mobile phase Inject 10 l of acetonitrile. Verify that no peak is detected in the chromatogram. Inject 10 l of standard solution for six times and record the chromatograms. Calculate the % RSD for retention times and peak areas for the six replicate injections.

Acceptance Criteria: % RSD calculated for Retention times and areas should
not be more than 1.0%

AUTO INJECTOR LINEARITY:Inject 20 ml methanol as a blank and record the chromatogram. Inject The standard solution (0.01mg/ml) for , 10 l, 20 l, 30 l, 40l.and 50l into the system and record the chromatograms. Take the areas for the concentrations of 10 l, 20 l, 30 l, 40l.and 50l and plot a graph for concentration (ml) Vs area counts.

Acceptance Criteria: Correlation coefficient should be 0.99 to 1.00

Calibration of GC:Calibration Frequency: Once in three months

Calibration procedure:Calibration of Flow rate accuracy Linearity of detector response & Injector Temperature calibration (column oven)

Calibration of Flow rate accuracy :Connect column one end at the injector port and the other end at the detector. Put on the carrier gas flow, and check for any leakages at both ends of the column. Connect the tubing of the digital flow meter. Set the flow rate at 3, 4 & 5ml/min respectively and measure the flow from the detector at each flow .

Acceptance criterion:
The accuracy of flow should not be more than 15%

Linearity of detector response & Injector:Chromatographic Conditions: Column : Detector : Column oven : Injector temperature : Detector temperature : Carrier gas : Flow rate : Injection Volume : DB-1 (15 m x 0.53 mm I.D) FID 130C 250C 250C Nitrogen / Helium 25 Kpa (58.3 cm /sec Linear velocity 1 l

Preparation of standard solution (10 m g of n-Dodecane , n-Tetradecane .and nHexa decane in 100 ml of n- Heptane) Preparation of level-1,2 & 3 solutions from stock solution Injection of level- 1,2 &3 solutions. When the chromatograms are recorded in the prescribed conditions, the order of elution is n-Dodecane , n-Tetradecane .and n-Hexa decane Calculate the % RSD of retention times & peak area responses of n-Dodecane , n-Tetradecane .and n-Hexa decane at each level

Acceptance criterion :
The % RSD of retention times of n-Dodecane , n-Tetradecane .and n-Hexa decane should be less than 2.0 at each level The % RSD of peak area responses of n-Dodecane , n-Tetradecane .and n-Hexa decane should be less than 5.0 at each level Calculate the mean peak area of responses of n-Dodecane , n-Tetradecane .and n-Hexa decane at each level Plot a curve of mean peak areas of each component at each level (y axis) against concentration (l ) of each level (x axis). Calculate the correlation coefficient

Acceptance criterion: The correlation coefficient should be more than 0.998

Temperature calibration (column oven) :

Set the column oven temperature at 40C,100C,150C,200C and 250C respectively and measure the temperature attained at each setting by placing the temperature probe inside the column compartment

Acceptance criterion:
The temperature attained inside the column compartment should be with in 2C for 40C and 50C to the set temperature and 5C for 100C,150C,200C and 250C to the set temperature

 Oven temperature control test Programmable Temperature Control test FID Sensitivity test

Oven temperature control test:

Keep the pre-calibrated sensors in oven Temperature is set at 500C, 1500C, 2500C. Waited till temperature is stabilized and recorded the minimum and maximum temperature in 10 min. Acceptance criteria 4.20C

Programmable Temperature Control test

Inserted the calibrated thermometer at the center position of the oven from the left side cover. Set temperatures: Column temperature, Injection temperature, Detectortemperature at 500C. Oven temperature: Initial temp. at 500C. Hold time of initial temperature about 1min. Temperature is programmed at rate of 130C/min. Final temperature of 620C is attained. Hold time of final temperature for 1min. Final temperature displayed. Acceptance limits 50C

Calibration of UV spectrophotometer:once in a month

Calibration procedure:Control of Absorbance Resolution power Limit of Stray Light

Control of Absorbance :Preparation of Potassium dichromate Solution Measure the absorbance of potassium dichromate solution at the wavelengths given below Calculate value of A (1% 1cm) for each wavelength A (1% 1cm) = Absorbance X 10000 / Weight of Potassium dichromate in mg. Acceptance criteria Wavelengths (nm) 235nm 257nm 313nm 350nm 124.5 144 48.6 106.6 122.9-126.2 142.4-145.7 47-50.3 104.9-108.2 A(1%1cm) Limit

Resolution power:Preparation of 0.02% v/v solution of toluene in hexane Record the spectrum of a 0.02% v/v solution of toluene in hexane in the range of 260 nm to 420 nm

Acceptance criteria:
The ratio of the absorbance at the maximum at about 269nm to that at the minimum at about 266nm is not less than 1.5.

Limit of Stray Light:Prepare a 1.2 % w/v solution of Potassium chloride in water Measure absorbance of the above solution at 198.0, 199.0, 200.0, 201.0, 202.0 nm using water as blank

Acceptance criteria:
Absorbance is greater than 2