Monday, 24 September 2018
Friday, 21 September 2018
SOP FOR OPERATING PROCEDURE OF ROTARY VACCUM EVAPORATOR
1.0
Objective:
The purpose of this SOP is to provide written procedure for
operation of Rotary Vacuum evaporator.
2.0
Scope:
This
SOP covers operating procedure of Rotary vacuum evaporator
3.0
Responsibility:
Junior Research officer, Research Officer:
Responsible for operation of the apparatus as
per procedure.
QA Officer/QA Manager: Review the
records and governing the document.
4.0 Procedure:
4.1
Connect the assembly as given in manual.
4.2 Fill water in water bath and start heating
so that temp of water should be 40 ± 2°C
4.3 Take extract in round bottom flask.
4.4 Connect to assembly so that part of the
flask is dip in water bath.
4.5 Start vacuum pump set the vacuum at 470
mm of Hg.
4.6 Set the revolutions at 100 rpm.
4.7
Dry the sample till evaporates to dryness and collects the residue with
suitable solvent in vial.
SOP FOR NITROGEN OXIDES (NOx)
1.0 Scope:
1.1 This standard prescribes a method
for measurement of nitrogen oxides (as NOx) in air.
2.0 Principle
and Applicability:
2.1 Nitrogen oxides as nitrogen
dioxide are collected by bubbling air through a sodium hydroxide solution to
form a stable solution of sodium solution of sodium nitrite. The nitrite ion
produced during sampling is determined colorimetrically by reacting the exposed
absorbing reagent with phosphoric acid, sulphanilamide, and N (I-naphthyl)
ethylenediamine dihydrochloride.
2.2 The method is applicable to
collection of 24 hour sample in the field and subsequent analysis in the
laboratory.
3.0 Range and
Sensitivity:
3.1 The range of the analysis is 0.01
to 1.5 µg NO2/ml. With 50 ml absorbing reagent and a sampling rate
of 200 ml/min for 24 hours, the range of the method is 20 to 740 µg/m3 (0.01 to
0.4 ppm) nitrogen dioxide.
3.2 A concentration of 0.01 µg oxides
of nitrogen ( as NO2/ml) will produce an absorbance of 0.005 using
1-cm cells.
4.0 Interferences:
4.1 The interference of sulphur
dioxide is eliminated by converting it to sulphuric acid with hydrogen peroxide
before analysis.
5.0 Apparatus:
5.1 Absorber – Polypropylene tubes 164
x 30 mm, equipped with polypropylene two port closures. Rubber stopper cause
high and varying black valves and should
not be used. A gas dispersion tube with a fritted end of porosity B (70 to
100 µm maximum pore diameter) is used.
5.2 Spectrophotometer – capable of
measuring absorbance at 540 nm. Band width is not critical.
6.0 Reagents:
6.1 Absorbing Reagents – Disolve 4.0 g
of sodium hydroxide in distilled water and dilute to 1000 ml.
6.2 Sulphanilamide – Dissolve 20 g of
sulphanilamide in 700 ml of distilled water. Add, with mixing, 50 ml of
concentrated phosphoric acid (85 %) amnd dilute to 1000 ml. this solution is
stable for a month if refrigerated.
6.3 NEDA Solution – Dissolve 0.5 g of
N (l-naphthyl) ethylenediamine dihydrochloride in 500 ml distilled water. This
solution is stable for a month if refrigerated and protected from light.
6.4 Hydrogen Peroxide – Dilute 0.2 ml
of 30% hydrogen peroxide to 250 ml with distilled water. The solution may be
used for a month if protected from light.
6.5 Standard Nitrite Solution – Dissolve
sufficient desiccated sodium nitrite [ NaNO2 (assay of 97 % or greater)]
and dilute with distilled water to 1000 ml so that a solution containing 1000
µg NO2/ml is obtained. The amount of sodium nitrite to use is
calculated as follows:
1.500
G = --------------- x 100
A
Where
G = amount in g of sodium nitrite;
1.500 = Gravimetric
factor in converting NO2 into sodium nitrites; and
A = assay, percent.
7.0 Procedure:
7.1 Sampling – Assemble; the sampling
train including the absorber, critical flow control device and pump. Add 50 ml absorbing reagent to the absorber.
Disconnect funnel, insert calibrated flow meter, and measure flow before
sampling. If glow rate before sampling is less than 85 % of needle calibration,
check for leak or change filter as necessary. Remove flow meter and replace
funnal. Sample for 24 hours from midnight to midnight and measure flow at the
end of sampling period.
7.2 Analysis – Replace any water lost
by evaporation during sampling. Pipette 10 ml of the collected sample into a
test-tube. Add 1.0 ml of hydrogen peroxide solution, 10.0 ml of sulphanilamide
solution, and 1.4 ml of NEDA solution with through mixing through mixing after
the addition of each reagent. Prepare a blank in the same manner using
interval, measure the absorbing reagent. After a 10 min colour-development
interval, measure the absorbance at 540 nm against the blank. Read µg NO2/ml
from the standard curve (see 8.2).
8.0 Calibration:
8.1 Sampling
8.1.1 Caligration of Flow
meter – Using a wet test meter and a stopwatch, determine the rates of airflow
(ml/min) through the flow meter at several ball positions. Plot ball positions
versus flow rates.
8.2.2 Calibration of Hypodermic needle – Connect the calibration flow meter, the needle to be calibrated,
and the source of vacuum in such a way that the direction of airflow through the needle is the same as in
the sampling train. Read the position of the ball and determine flow rate in
ml/min from the calibration chart prepared in 8.1.1. Reject all needles not having flow rate of 190 to 210 ml/min
before sampling.
8.2 Calibration Curve – Dilute 5.0 ml
of the 1000 µg NO2/ml solution to 200 ml with absorbing reagent.
This solution contains 25 µg NO2/ml. Pipette 1, 2, 5 and 15 ml of
the 25 µg NO2/ml solution into 50, 50, 100 and 250 volumetric flasks
and dilute to the mark with absorbing reagent. The solutions contain 0.50,
1.00, 1.25, and 1.50 µg NO2/ml respectively. Run standards as
instructed in 7.2. Plot absorbance versus µg NO2/ml.
9.0 Calculation:
9.1 Calculation
the volume of air samples as follows:
F1 + F2
=
------------------------- x T x 10-6
2
where
V = volume of air sampled, m3;
F1 = measured flow
rate before sampling, ml/min;
F2 = measured flow
rate after sampling, ml/min; and
T = time of sampling, min.
9.2 Calculate the concentration of
nitrogen dioxide as follows:
(
µg NO2/ml) x 25
Mass
of nitrogen dioxide in µg per m3 = -------------------------------
V x 0.35
Where
25 = volume
of absorbing reagent used in sampling, ml;
V = volume of air sampled, m3;
and
0.35 = overall
average efficiency.
9.2.1 If desired,
concentration of nitrogen dioxide may be calculated as ppm NO2.
NO2,
ppm = ( µg NO2/m3) x 5.32 10-4 SOP FOR OPERATING PROCEDURE OF HIGH PERFORMANCE LIQUID CHROMATORGAPHY SYSTEM SHIMADZU (Model: LC 2010CHT).
1.0
Objective:
To
provide written procedure for operation of High Performance Liquid Chromatography system
EQU-INS-201.
2.0
Scope:
This SOP covers operation of
High Performance Liquid Chromatography system make: Shimadzu (Model: LC 2010CHT).
3.0
Responsibility:
Jr. Research Officer, Research Officer: Responsible for operation,
Calibration and maintenance of the instrument as per procedure.
Head of Department: Responsible for calibration and maintenance,
timely as per schedule.
QA Officer/ QA Manager: Review the record and governing
the document.
4.0 Procedure:
4.1 Ensure
that the instrument is visibly clean and free from dust.
4.2 Starting UP and Connecting
Instruments and Class –VP Software:
4.2.1
Turn on the instruments.
4.2.2 After confirming that the instrument is
started up, turn the PC power ON, and select and click on Start icon, Select
programs and chromatography and click on ‘CLASS VP’. OR double click on ‘CLASS VP’
icon on desktop. Display shows Class VP dialog box.
4.2.3
Select and double click on the instrument
icon in the Class VP dialog box.
4.2.4
Enter a user name ‘System’ and password
‘2001’; to log in to ‘CLASS-VP’, the instrument window opens.
4.3 Click on pump
icon or and fed the require flow rate and concentration of solvent.
4.4 Purging Mobile phase and Rinse Solution:
4.4.1 Click on the ‘PURGE’ icon on the control
toolbar. Select the flow lines to be purged and set purge time for each.
4.4.2
Clicking on the purge button
display a window showing the progress of purge, and the auto purge starts.
4.4.3
If stop the purging click
‘Mobile Phase Stop’ button.
4.4
Click on ‘PUMP’ icon into
instrument window; Select the mode of pump as Isocratic flow or low pressure
Gradient, feed the flow rate and concentration of solvent in %, adds maximum
pressure and minimum pressure value.
4.5
Create a New Method File or
Modify Method:
4.6.1 Choose Open or ‘New’ option from the method
in File menu in the instrument window, Method dialog box display on the screen.
4.6.2 Select the commands in the Method menu from
the Option tab, Select and click on Properties, Method properties dialog box
display, feed the require parameters, then click on ‘OK’ button.
4.6.3 Select and click on Integration Events in
method menu, Integration Events dialog box display, feed the require
parameters, and then click on ‘OK’ button.
4.6.4 Select and click on Peaks/Groups in method
menu, Peaks/Groups dialog box display; add Component name, Retention time etc.
and then click on ‘OK’.
4.6.5 Select and click on Advanced, Advanced
method option dialog box display, Select Component name, Retention time, Area,
Asymmetry, and Resolution etc.
4.6.6
Select and click on Instrument Setup in
method menu, Instrument dialog box display, and click on ‘PUMP’ button.
4.6.7
Select the mode of pump as Isocratic flow or
low pressure Gradient, feed the flow rate and concentration of solvent in %,
adds maximum pressure and minimum pressure value.
4.6.8
Click on ‘Oven’ button, add temperature maximum
up to 60° and add Oven temperature.
4.6.9 Click on ‘Detector’ button, Select D2 lamp,
Polarity, Cell temperature low, add wavelength on Channel 1 and require for
Channel 2, add sensitivity and select Acquisition On, add require Run time.
4.6.10
Click on ‘Controller’ and select Degasser.
4.6.11 Click on ‘Time
Program’ and select module (Pump, Oven, Detector etc.) add require time and
program.
4.6.12
Select and click on System
Suitability in method menu, System Suitability dialog box display, select the
require parameters i.e. Area, Retention time, Asymmetry, Resolution etc. and
then click on ‘OK’.
4.6.13
After completion all the parameters save
method from file menu. Enter or Select path and full file name.
4.7 Create a New Sequence File or
Modify Sequence :
4.7.1
Choose Open or ‘New’ option from the Data in
File menu in the instrument window, Data dialog box display on the screen,
create a new folder and give name of the folder.
4.7.2
Choose Open or ‘New’ option from the
sequence in File menu in the instrument window, sequence dialog box display on
the screen.
4.7.3
In Run information add Sample ID name,
Select method, Select Data path, and give Data file name.
4.7.4
Add Amount values as per requirement.
4.7.5
In Auto sampler feed the parameters, start
vial, end vial, injection volume and Repetitions per run, and then click on
‘OK’, sequence file display on the screen.
4.7.6 In sequence file feed parameter sample
identification, vial number, injection volume respectively.
4.7.7
After completion all the parameters save
sequence from file menu. Enter or Select path and full file name.
4.7.8 After completion all parameters, right click
of the mouse, select and click on start sequence.
4.8 Creating Report Templates:
4.8.1 Click on the Edit Custom Report button or
Select and click on Custom Report, Custom Report dialog box display.
4.8.3 Select print in file menu, and give print
command for printing.
4.9 Opening of Instrument Off Line:
4.9.1 Select and double click on the
instrument off line icon in the Class VP dialog box.
4.9.2
Enter a user name ‘System’ and password
‘2001’; to log in to ‘CLASS-VP’, the instrument off line window opens.
4.10 COLUMN FLUSHING :
4.10.1 After completion of analysis clean the
column by the following solvents as mobile phase.
4.10.2 For reverse phase columns:
4.10.2.1
Flush with the same mobile
phase, which was need for analysis for 15 minutes with 1.0 ml flow rate.
4.10.2.2
Flush with water for 30
minutes.
4.10.2.3 Then flush with methanol for 15 minutes at
a flow rate of 1.0 ml / minute.
4.10.3 For normal phase columns:
4.10.3.1 Flush with the same mobile phase which was
need for the analysis for 15 minutes at a flow rate of 1.0 ml / minute.
4.10.3.2 Then flush with n -Hexane at a flow rate of 1.0 ml / minute.
4.10.4 Change over from Reverse phase to Normal phase :
4.10.4.1 After the analysis is over flush the column
with water for 30 minutes, followed by Methanol for 15 minutes.
4.10.4.2
Remove the reverse phase column
(C18 or C8) and attach a dead volume instead of
4.10.4.3
column.
4.10.4.3 Now flush the system with acetonitrile
followed by chloroform and n -hexane (all 15 minutes each).
4.10.4.4 Remove dead volume and fix a normal phase column (e.g.
silica)
4.10.4.5 Continue flushing for 15 minutes.
4.10.4.6 Maintain the reverse phase column in
methanol.
4.10.5 Change over from normal to reverse
phase:
4.10.5.1 After the analysis is over flush the
columns with n- hexane for 15 minutes.
4.10.5.2 Remove the normal phase column (C18 or C8)
and attach a dead volume instead of
column.
4.10.5.3 Flush the system with chloroform followed
by acetonitrile and methanol for 15
minutes.
4.10.5.4 Now fix the reverse phase column and flush
the column for 15 minutes.
4.10.5.5
Continue the analysis in
reverse phase. Store the normal phase column in n -hexane.
4.11 Enter the details in HPLC usage
logbook as per given in Annexure –1.
5.0 Safety precaution:
5.1
After analysis run the system wash with water for 30 minutes if buffer
solution used in mobile
phase followed by methanol for 15 minutes.
6.0 Routine maintenance:
6.1 Clean the instrument by dry cloth.
6.2 Clean the system with hot water
without connecting the column. Than methanol and
finally with water.
6.3
Sonicate the suction filter with 2 M
nitric acid and than water and than water.
SOP FOR OPERATING PROCEDURE OF GAS CHROMATOGRAPHY AGILENT 6890N
1.0
Objective:
The objective of this Operating
procedure is to lay down the procedure for Operating the Gas Chromatography
Agilent 6890N.
2.0 Scope:
This procedure covers operation
of Gas Chromatography.
3.0
Responsibility:
Jr.
Research Officer, Research Officer: Responsible for operation and maintenance
of
the
instrument as per procedure.
Head of Department: Responsible for maintenance, timely as per schedule.
QA Officer/QA Manager: Review the records and governing the document
4.0 Procedure:
4.1
I.
Ensure that the instrument is visibly clean and free from dust.
II.
The room
temperature should be maintained between 10 to 30°C and relative humidity
Should be maintained between 30 to
80 %
III. Avoid direct exposure to air-blow from air
conditioners.
IV. Avoid exposure to direct sunlight and
vibrations.
V. Avoid from equipment which generates
strong magnetic field.
4.2
Start UP Procedure:
4.2.1
Affix the require column (Capillary or packed) in to the column oven.
4.2.2
Open the valve of gas cylinder of Z - air, Hydrogen and carrier gas
(Nitrogen or Helium) and checked the pressure and leakage with 50 % v/v
Isopropyl Alcohol solution.
4.2.3
Now Switch ON the Mains and Instrument. Switch on the computer.
4.3 Setting Instrument Parameters:
4.3.1 After Switch On instrument will display
Oven Off message. Press oven key and press on key. The instrument started
temperature increase of previous set temperature value.
4.3.2 Press Oven key, Add initial temperature,
initial time and Rate (if required) with numerical key and press Enter key to
confirm the vale.
4.3.3 If adding rate 1 the next oven program
display, feed the required temperature, time and rate and then press Enter key
to confirm the value. If column (Oven) temperature is Isothermal condition the rate is off.
4.3.4 Set the Oven Final Temperature below 20°
C of Maximum Column temperature or as per procedure requirement.
4.3.5 Press ‘Back inlet’ key select mode split or
split-less, add require inlet temperature with numerical key and press ‘Enter’
key to confirm the value.
4.3.6 Press ‘Front Det.’ key, add require detector
temperature with numerical key and press ‘Enter’ key to confirm the value. Now
increase the temperature, if temperature comes out at 150° C message display
‘FLAME OUT’.
4.3.7 Press ‘Front Det.’ Key and select Flame with
▲ and ▼ key select flame on with press ‘On’ key. Igniting message display and
ignite the flame with gas lighter on detector side. The display shows the flame
indicator in a numerical value.
4.3.8 Press ‘Status’ key and check the fed
parameters again with ▲ and ▼ key to confirm the value. Now instrument is
ready, the message will display “Instrument ready for Injection’.
4.3.9 Press “Signal 1” key select Zero with▲ and ▼
key, press ‘On’ key to flame numerical value should be zero and base line
detection started to zero mv.
4.3.10 Now inject the solution with syringe. If
used manual operation press ‘START’ key immediately after inject the solution.
4.4
Software Operation:
4.4.1 Double click on the instrument1 (online) &
instrument1 (offline) icon from the desktop;
create new method
from instrument1 (offline) and load method in instrument1 (online)
from
file menu. Apply the chromatographic condition and enter ok. And view the run
signal
window, After completion of the run, and open the instrument offline select the
file
and]
integrate the peak using integration menu, go to calibration task icon and
enter the
component name
and click on ok, then print the chromatogram by clicking print option
from file menu.
4.4.2 Condition the column at
280°C for 10 min. Enter the programming condition of the sample through micro
syringe, when G.C shows ready signal.
4.4.3 Cool the G.C after
completion of the analysis & switch it off. Shut down the computer,
mains power supply. Close the gas
supply from the distribution panel & from cylinder.
5.0 Routine Maintenance:
5.1 Clean the all out side part of
instrument.
5.2
Clean the injector and injector liner. Change the glass wool of injector liner.
Change the
septa of injector port if required.
5.3
Check
the all connection of gases for leakage.
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