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SOP for the sampling and analysis of Benzo (a) Pyrene in Ambient air

(Solvent Extraction and Analysis on GC)


1. Principle Benzo (a) Pyrene sampled in ambient air in Whatman glass Fiber using
Respirable dust Sampler and Extracted in Organic Solvent and Analyzed on GC-FID.
2. Apparatus/ Instrument Required:
a)
b)
c)
d)
e)
f)

Respirable Dust Sampler.


What man glass fiber (EPM 2000 Filter paper).
Rotary Evaporator.
Ultrasonicator
Silica-Gel Column 200mmL,5mm Internal diameter
GC-FID With capillary Column(Length25m,0.320mm Internal diameter & 0.17 m Film
Thickness)
g) Syringes,1-10l
h) Micro-Pipettes,500-1000 l
i) Flask & Beakers, 5,10,25,50 &250ml.

3. Reagents and Chemicals:


a. Standard Benzo (a) Pyrene (PAHs high purity)
b. Toluene (Ultra residue grade)
c. Cyclohexane (Ultra residue grade)
d. Activated Silica Gel (60-100 meshes)
e. Glass Column for Clean up
4. Procedure: Ambient Air Sampling is performs for PM10 using Respirable Dust sampler for 8
hrs. Using Pre Weighted filter paper Whatman glass fiber EPM-2000 and dust cup.
After determined of PM10 and total suspended Particulate Matter by post weighing the
Entire dust is added to 250 ml Beaker along with filter paper and is extracted with 50ml of
Toluene 3 times after 30 minutes ultra sonication each time.
Then the extract is collected in a conical flask (250ml) for filtration.

Toluene extract is filtered through Whatman No. 41containing 2gm of anhydrous sodium sulphate
For removal of moisture and Particles & it is evaporated to 2ml in rotary evaporator.
To clean up the impurities, pass 2 ml sample through silica gel glass column (60-80mesh, and 200250mm10mm with Teflon stopcock).

The conditioned silica column filled with silica with toluene is used for further clean up.
After loading 2ml of Toluene reconstituted extract it is eluted & using 5 ml cyclohexane three
times (17 ml total).

The elute is evaporated to nearly dryness in rotary vacuum evaporator and finally reconstituted
to 1.0 ml in Toluene and transfer into 2ml vial for Analysis.
GC condition:Column- capillary Column (Length25m, 0.320mm Int. dia & 0.17m Thickness)
Temperature programming
Injection part: 320C
Detector: 320C
Column/oven: 140C (Hold 3 min)
Ramp 1 @6C /Min 250C (Hold 6 min)
Ramp 2 @10C /min 300C (Hold 5 min)
Total Run Time: 36 Min Approx.
Injection: Volume 1 l, 5l
Calculation:
Conc. of Banzo(a)Pyrene,in ng/m3,of each analyte in the air sampled C=
Conc. in (ng/l) of each analyte, Cs x 1000l (Final Vol of extract,Ve, l)
Volume of Air sampled, Vs in (m3)
Calculation of,
Conc. in (ng/l) of each analyte, Cs= (As Cis)/ (AisRF)
Where,
As= area count of analyte sample/peak
Ais=area count of internal standard/peak
Cis=conc. of internal standard.
Calculation of,
RF= (As Cis)/ (Ais Cs)
Where,
As= area of analyte (sample/standard) to be measured
Ais= area of internal standard
Cis= conc. of internal standard, (ng/l)
Cs=conc. of analyte (sample/standard) to be measured (ng/l)

Volume of Air sampled, Vs in (m3) =V (Pa (ambient pressure)/101kPa) [298/ (273+Ta


(Ambient Temperature)]

Reference :( 1) Guideline for sampling and analysis of Benzo (a) Pyrene & other PAH in Ambient
Air (Solvent Extraction & GC- FID or GC- MS or HPLC/UV- Fluorescence Detector)
(2) Polynucler Aromatic hydrocarbon (PAHs) in Air Particulate as per IS: 5182
(Part- 12):2004

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