READ AND REVIEW ANY APPLICABLE MANUFACTURER/VENDOR SAFETY INFORMATION BEFORE
DEVELOPING STANDARD OPERATING PROCEDURE AND PERFORMING WORK.
PI Name: Sergey Nizkorodov
Date Modified: February 17, 2016
Name of Work Unit: Aerosol Photochemistry Group
Standard Operating Procedure (SOP) for
Carbonyl Test
#1
Scope of Work/Activity: Describe the experiment purpose/scope.
- Identify all apparatus that will be used, and associated requirements.
- List special equipment (X-ray generators, lasers, furnaces, etc.) that will be used for
the project.
- Identify measurement and test equipment, apparatus operating conditions,
Include process flow diagrams, pressures, and required maintenance procedures as
appropriate.
Purpose/Scope:
 To trace quantities of carbonyl compounds in the range from 0.5 to 50 µg.
Apparatus/Equipment: (All glassware needs to be rinsed with water and finally with
Carbonyl-Free Methanol. Do not use acetone to dry the glassware.)
 Volumetric flasks
 Glass beakers
 Pipette
 Shimadzu UV-Vis Spectrometer to measure the absorbance of the carbonyl
solution. (Look at Shimadzu SOP)
 Absorption Cells, 1-cm
Reagents:
 2,4-dinitrophenylhydrazine (10 mg)
 Potassium Hydroxide (10 g)
 Carbonyl-free Methanol (400 mL)
 2-Butanone (used for calibration curve) (50 µL)
 Hydrochloric Acid (400 µL)
 Need at least 200 µL of unknown compound containing carbonyl
#2
Specific Safety and Environmental Hazards: State the specific hazard and
consequences if procedure not followed to person, environment, or property.
2,4-Dinitrophenylhydrazine: flammable, toxic.
Potassium Hydroxide: Corrosive. Water-Reactive. Harmful if swallowed. Causes
severe eye and skin burns. Causes severe digestive and respiratory tract burns.
Carbonyl-Free Methanol: Flammable liquid and vapor. Harmful if swallowed, inhaled,
or absorbed through the skin. Causes eye, skin, and respiratory tract irritation.
2-Butanone: Highly flammable. Incompatible with oxidizing agents, bases, strong
reducing agents. Protect from moisture.
Hydrochloric Acid: Corrosive. Liquid and mist cause severe burns. May be fatal if
swallowed or inhaled.
#3
Check MSDS prior to working with the chemicals.
Describe in detail how the hazards will be controlled.
a. Identify the Engineering Controls (e.g. interlocks, shielding), Standard Operating Procedures, or
Personal Protective Equipment (e.g. respirators, gloves;) that will be employed to reduce hazards
to acceptable levels.
 Wear gloves and goggles at all times.
 Work in the fume hood to minimize inhalation of hazardous vapors
#4
#5
#6
b. Address emergency shutdown procedures.
In an event of an emergency:
 Report leaks, spills, suspicious odors to supervisor immediately.
 If hazardous vapor is inhaled, move immediately to fresh air
 Remove potential sources of ignition.
 Evacuate area.
 Stay away until given the “all clear.”
 Refer to MSDS for specific spill and leak information.
 Know what chemical or chemicals you were working with.
.
Designated Area: Indicate the designated area for performing this process in the
laboratory.
In the fume hoods. Preferably next to the UV-Vis to have easy access when measuring
the absorbance.
Personal Protective Equipment (PPE): State the personal protective equipment selected and
required. Examples: safety spectacles, work gloves, respiratory protection, steel toe shoes.
Personal protective equipment includes:
 Goggles must be worn by all people in the vicinity of the chemicals.
 Closed toe shoes to prevent any accidents that may occur.
 Rubber gloves when working with chemicals.
Important Steps to Follow: List the specific sequence staff should follow to avoid
hazard.
Refer to the procedures provided at the end of this document.
#7
Emergency First Aid Procedures:
a. Describe immediate medical treatment required in case of personnel exposure.
-Complete online incident report form at www.ehs.uci.edu
Refer to PPE in #3b.
#8
Training & Competency Requirements: Describe necessary training and demonstration
of competency for performing the hazardous operation.
 Pass the Safety Training Assessment courses online. (including Laboratory
Core Safety, Fire Extinguisher Safety, and Hazardous Waste)
 Read this SOP
 Read UV-Vis and Shimadzu SOP
Identify waste stream and disposition of unused stock of chemicals
List concentrations and amounts of hazardous wastes or emissions and control
measures.
Additional guidelines regarding hazardous waste at :
http://www.ehs.uci.edu/programs/enviro/
 Surplus chemicals will be disposed of as hazardous chemical waste in the organic waste
bucket.
Decontamination and spill clean-up procedures
 Remove any contaminated clothing
 Notify other workers in the area of the spill and control traffic through area.
 Put on gloves and cover spill area with paper towels.
 Pour disinfectant over towels from edges of spill to center, be carefully not to splatter.
 Decontaminate all objects in spill area.
 Allow 30 minutes of contact time.
 Pick up any sharps, including broken glass, with forceps and place in sharps container.
 Use dustpan to recover any shards of broken glass in contaminated liquid.
Decontaminate dust pan.
 Wipe area with disinfectant and clean paper towels and put in biohazard bag.
 Mop if spill is on floor.
#9
#10
As the Principal Investigator, it is your responsibility to ensure that all individuals listed in this protocol is
taught correct procedures for the safe handling of hazardous materials involved in this study. It is also your
responsibility to assure that your personnel attend Lab Core Safety Training and other applicable safety training
courses.
Both PI and all persons associated with the protocol must sign the following acknowledgement:
I have read, asked questions, and understand the hazards of and safe working procedures for the
activity/materials described herein.
PI Signature:
DATE
Other Personnel:
Name/ Signature
DATE
Name/Signature
DATE
Name/Signature
DATE
Summary of Operating Procedures for Carbonyl Test
Last updated on February 17, 2016
PPE required
 Goggles
 Gloves
 Close toe shoes
(All glassware needs to be rinsed with water and finally with CarbonylFree Methanol. Do not use acetone to dry the glassware!)
Preparation of Reagents:
Carbonyl-Free Methanol: Have to purchase from Sigma Aldrich.
Potassium Hydroxide Solution:
1. Dissolve 10 g in 20 mL of water in a 100 mL volumetric flask.
2. Cool and dilute to 100 mL with methanol.
2,4-Dinitrophenylhydrazine (Solution is unstable and must be discarded after two weeks)
1. Dissolve 10 mg of 2,4-Dinitrophenylhydrazine in 5 mL of carbonyl-free methanol containing 400 µL of
hydrochloric acid.
2. Dilute to 10 mL with water in a 10 mL volumetric flask.
Calibration:
1. Add 34.7 µL in a 100 mL volumetric flask containing 50 mL of carbonyl-free methanol.
2. Fill the rest of the flask with carbonyl-free methanol.
3. Prepare a series of aliquots with concentrations between 7.74E-5 and 3.87E-4 M.
Procedure:
1. Dilute the unknown carbonyl solution. May require a test trial to obtain a rough estimate of the dilution
factor. Make sure that the absorbance is within the calibration curve. (For CH3OOH, dilute solution to ~
4E-3 M)
2. Transfer 800 µL of this stock solution into a 10 mL glass-stoppered volumetric flask.
3. Transfer 800 µL of each aliquot solution of 2-butanone to respective 10 mL glass-stoppered volumetric
flasks.
4. Transfer 800 µL of carbonyl-free methanol to a 10 mL glass-stoppered volumetric flask for use as a
reagent blank.
5. To each flask, transfer 800 µL of the 2,4-dinitrophenylhydrazine solution. Stopper and let sit for 30 ± 2
minutes.
6. Dilute to the mark with potassium hydroxide solution. Stopper and mix well.
7. At 12 ± 1 minute of adding the potassium hydroxide solution, measure the absorbance at 480 nm in a 1cm glass cuvette. (Read UV-Vis and Shimadzu SOP prior to using the spectrometer)
8. Use the calibration curve to calculate the concentration of the unknown carbonyl solution.