Table of Contents
1.0 OBJECTIVE
To lay down the procedure for operation and calibration of UV visible spectrophotometer (Make: Shimadzu, Model No.: UV-1900 I).
2.0 SCOPE
This SOP is applicable for operation and calibration of UV visible spectrophotometer in quality control department.
3.0 RESPONSIBILITY
QC – Officer /Sr. Officer/ Executive and above.
4.0 ACCOUNTABILITY
Head-QC
5.0 PROCEDURE
5.1 Operating Procedure for photometric mode:
5.1.1 Ensure the instrument is clean & free from dust
5.1.2 Switch “ON” the mains & switch “ON” the computer and UV-visible spectrophotometer
5.1.3 Press on “P.C CONTROL” after initialization is completed on Instrument.
5.1.4 Click on lab solution software installed on desktop of your P.C.
5.1.5 Select the method you want to use.
5.1.6 If you want to use “PHOTOMETRIC MODE “, click on it.
5.1.7 Click on “CONNECT” key and instrument control will display on screen.
5.1.8 Click on “READ” to select the method and now go to file name.
5.1.9 File name for products shall be named as follows
“Product Name/Calibration_TestName_DDMMYY_Version No.”
For Example,
“CEFT_ID_250124_A”, “Calibration Control of wavelength_250124_A”.
Where,
CEFT is first four digits of product of Ceftriaxone.
ID is test name of product for Identification
250124 is date month and last two digit of year.
5.1.10 If in the first sequence any incident occurs, then new sequence shall be named as
“CEFT_ID_250124_A1”.
5.1.11 The sequence for same Product/calibration, same test but having different batch no./wavelength then the sequence will be “CEFT_ID_250124_B”, “Calibration Control of wavelength_250124_B”.
5.1.12 Put blank on both the side of reference cell holder and sample cell holder click on “AUTO ZERO” in case of Single Wavelength and in case of more than one wavelength click on “BASELINE CORRECTION” displayed on “INSTRUMENT CONTROL”.
5.1.13 After completion of “AUTOZERO” or “BASELINE CORRECTION”.
5.1.14 Fill photometric table.
5.1.15 Place the cuvette fill with sample on sample side holder Close the sample compartment in uv and take the readings by clicking on unknown sample.
5.1.16 After completion of analysis data file is automatically saved.
5.1.17 Now go to edit print form and open to select to template (Photometric Template Sigma) and take print.
5.1.18 Note: The photometric uses of take a one and multipoint reading of known wavelength of unknown absorbance.
5.2 Operating Procedure for spectrum mode:
5.2.1 Now select on “SPECTRUM” menu.
5.2.2 Click on “CONNECT.”
5.2.3 Click on “INSTRUMENT CONNECT”.
5.2.4 Click on “READ” to select the method and now go to file name.
5.2.5 File name for products shall be named as follows point no. 5.1.9 to 5.1.11.
5.2.6 Now go to sample name, write sample/product name in sample name column and in sample ID column write to batch no. of product/ material.
5.2.7 Now Place both cuvettes filled with solvent or blank in sample compartment and click on baseline correction.
5.2.8 After doing baseline correction if Absorbance readings will show “0.000” then it is ok.
5.2.9 Now place the cuvette on sample side filled with sample and close sample compartment in uv.
5.2.10 Click on start.
5.2.11 Let the graph be completed
5.2.12 After that click on active shown above the graph plotted then right side of the corner there is threshold put the value 0.00 in it tick on both “PEAK and VALLEY” and click on “EXECUTE.”
5.2.13 For the saving the data for future use click on file click on “SAVE AS DATA” and data save as Win(C:)►UV Vis-Data►Data and now Open file the go to edit print form and open to select to template (Spectrum Template Sigma) and take print.
To Read this sop click here SOP for Cleaning Of Microbiology Lab.
5.3 Precautions:
5.3.1 Do not open sample compartment during initialization, baseline correction, Auto zero and sampling Measuring Modes.
5.3.2 Before placing, the Cuvettes in the sample holder clean the outer surface of cuvettes with soft tissue papers or lint free clothes and do not hold the cuvettes from its transparent optical surface.
5.3.3 Operate the instrument after proper initialization.
5.3.4 Do not operate the instrument without instrument connect with system.
5.4 Method Creation in Photometric mode:
5.4.1 Switch “ON” the mains & switch “ON” the computer and UV-visible spectrophotometer.
5.4.2 Press on “P.C CONTROL” after initialization is completed on Instrument.
5.4.3 Click on lab solution software installed on desktop of your P.C.
5.4.4 Click on Photometric mode.
5.4.5 Click on “CONNECT” key and instrument control will display on screen.
5.4.6 Click on “PARAMETER EDIT”
5.4.7 Click on “REGISTRATION OF WAVELENGTH”
5.4.8 Put parameter, put wavelength, click on “ADD” and then click on O.K
5.4.9 Close after creating new parameter file, put method name as follow:
“Product Name /Calibration _Test Name at wavelength”.
Example:
“Calibration Control of absorbance at 430nm”.
5.5 Method Creation in Spectrum mode:
5.5.1 Now select on “SPECTRUM” menu.
5.5.2 Click on “CONNECT.”
5.5.3 Click on “INSTRUMENT CONNECT”.
5.5.4 Click on “PARAMETER EDIT.”
5.5.5 Select the scan range as per your requirement, scan speed medium, data interval 0.5 nm
5.5.6 Click on “close after creating new parameter file.”
5.5.7 Enter the file name as follow point no.5.4.9.
5.6 Calibration:
5.6.1 Operate the instrument as per the procedure given below:
• Control of Wavelength
• Control of Absorbance
• Limit of Stray light
• Resolution power
• Second Order Derivative
• Solvent