KANSAS = DEPARTMENT OF AGRICULTURE LABORATORY

KANSAS DEPARTMENT OF = AGRICULTURE=20 LABORATORY

Preparation of Tank Mix = Samples For=20 Gas Chromatography or High Pressure Liquid=20 Chromatography

1. =20 Purpose

To=20 determine by way of Gas Chromatography or High Pressure Liquid = Chromatography=20 the percentage of an analyte in a tank mix.

2. =20 Scope

This=20 SOP concerns the typical preparation of tank mixes to be run by either = the HPLC=20 or GC.

3. =20 References

=20 3.1 =20 National=20 Enforcement Investigation Center

=20 3.2 =20 Official Methods of Analysis of AOAC International, = 17^th=20 Edition

=20

4. =20 Safety and Hazardous Waste

4.1 = Standard=20 personal protective equipment for the laboratory shall be used. MSDS = information=20 may be accessed in the laboratory file. Any hazardous waste shall be = disposed of=20 according to the laboratory SOP KDAL-AM-WD-1. Usual good laboratory = practices=20 shall be adhered to during the procedures.

5. =20 Reagents / Supplies / Equipment

=20 Ring Stand

=20 Separatory Funnel 250ml

=20 Beakers

=20 Graduated Cylinders

=20 Round Bottom Flask 250ml

=20 Funnel

=20 Glass Wool

=20 Rotary Evaporator

=20 Rotary Evaporator Stand

=20 Hot Plate / Water pan

=20 Water Bottle

=20 Sodium Sulfate

=20 Methylene Chloride

=20

6 =20 Outline of Procedure

6.1 =20 Pre-preparation

6.2 = Standards=20 Book and Worksheet

6.3 = 50/50=20 dilutions

6.4 = Filtered to=20 Vial

6.5 = Phase=20 Over

6.6 =20 Calculations

7 =20 Specific Procedure

7.1 =20 Pre-=20 Preparation

7.1.1 Obtain the sample = according to SOP=20 KDAL-PL-CC-3.

7.1.2 Sample is kept in locked = cabinet in=20 Formulations room.

7.1.3 Pesticide Formulations = Worksheet is=20 in file on top of counter by the east wall.

7.1.4 Determine appropriate=20 solvent.

7.1.5 Obtain standard in = standards=20 room.

7.2 = Standards=20 Book and Worksheet

7.2.1 Record information = regarding the=20 preparation of the standard.

7.2.2 Record what was done with = the=20 sample on the Pesticide Formulations Worksheet.

7.2.3 Record any dilutions or = any notes=20 about sample or standard in the note section on worksheet.=20

7.3 = 50/50=20 Dilutions

=20

7.3.1 =20 Sample = goes on=20 GC.

7.3.2 Shake sample to make=20 uniform.

7.3.3 Weigh 12.5 grams of = sample into 25=20 ml volumetric flask.

7.3.4 Go to volume with = solvent. Mix well=20 and vial.

7.4 = Filter to=20 Vial

=20

7.4.1 =20 This = sample=20 would typically go on HPLC.

7.4.2 Supplies are in drawer = under=20 balance.

7.4.3 Take a 3cc syringe and = put a 0.2=20 micron filter on the end. Place end of filter in vial, pull plunger from = syringe=20 and pour sample into syringe.

7.4.4 Replace plunger and = inject sample=20 through the filter into the vial and cap.

7.5 = Phase=20 Over

=20

7.5.1 Set up stand for 250ml = separatory=20 funnel in hood.

7.5.2 Weigh up 12.5 grams of = sample in=20 beaker.

7.5.3 =20 Pour = sample into=20 separatory funnel and add small amounts of sodium sulfate, shaking = funnel in=20 between each addition. Keep repeating until sample becomes=20 opaque.

7.5.4 Pour 20 ml methylene = chloride and=20 shake.2 min. Let separate into layers.

7.5.5 Put a powder funnel = packed with=20 glass wool in a 250ml round bottom flask.=20

7.5.6 =20 Pour = sodium=20 sulfate over glass wool until about an inch deep.

7.5.7 Let bottom layer from = separatory=20 funnel run through funnel into round bottom flask. Repeat two more=20 times.

7.5.8 Set up hot plate and = rotary=20 evaporator in hood.

7.5.9 Rotary evaporate to=20 dryness.

7.5.10 Decide on solvent and final volume = and mix=20 well.

7.5.11 Vial sample according to what = instrument you=92re=20 going to use.

8. =20 Calculations

8.1 = Pesticide=20 Formulation Worksheet is in Excel.

External=20 Standard Tank Mix Calculator

Fipronil

Sample=20 Number

06-0078

Std = Resp

1701015

1695748

Standard=20 Average Response

1696796

1687081

Sample=20 Average Response

1821648

1703340

Standard=20 Conc.

0.6

mg/mL

Sample=20 Weight

163.2

Standard=20 Volume

Sam = Resp

1701710

Sample=20 Volume

1941585

Concentration=20 Percent

9.867

%=20

98.675

mg/g

8.2 = Enter the=20 following information into the spreadsheet

=20 =20 8.2.1 = Standard=20 Concentration

=20 =20 8.2.2 Sample=20 Weight

=20 =20 8.2.3 Sample=20 Volume

=20 =20 8.2.4 Std = Resp-Area of=20 Standard

=20 =20 8.2.5 Sam = Resp-Area of=20 sample