SPE Method Development for Pesticides

CASE STUDY
SPE Method Development for Pesticides

OBJECTIVE
The objective of this work was to develop a suitable SPE method for pesticide standards, to include
Aldicarb Pestanal Standard, Aldicarb-Sulfone Pestanal Standard and Oxamyl Pestanal Standard.

CONCLUSIONS
The resulting LCMS chromatograms showed good retention of the compounds and revealed that
methanol with 100 mM ammonium acetate is an effective elution solvent. 94% or greater recovery was
observed for all three pesticides using methanol with 100 mM ammonium acetate. Acetonitrile with 10
mM ammonium acetate also exhibited good results, with 90% or greater recovery in the first elute step.

Read the following report to see the full analysis.

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Final Report
Company Name

Date:

Released by:
Dr. Mark Jordi
President
Jordi Labs LLC

Report Number: J####

Company Name Confidential

Date

Client Name
Company Name
Address

Dear Valued Client:

Please find enclosed the test results for your samples described as:

1. Aldicarb Pestanal Standard

2. Aldicarb-Sulfone Pestanal Standard
3. Oxamyl Pestanal Standard

The following tests were performed:

1. Solid Phase Extraction (SPE) Method Development

2. Gas Chromatography Mass Spectroscopy (GCMS)
3. Liquid Chromatography Mass Spectroscopy (LCMS)

Objective
The objective of this work is to develop a suitable SPE method for the pesticide standards
provided.

Aldicarb-Sulfone Aldicarb Oxamyl

Summary of Results
SPE was performed using a Jordi Labs Hydroclean RP-200mg cartridge, part number
530SPE-006-00200. The following conditions were applied:

Step Conditions Conditions

Starting Soln. 0.5 mg/ml of each standard in water 0.5 mg/ml of each standard in
water
Conditioning 2ml Methanol ► 2ml DI Water 2ml Methanol ► 2ml DI Water
Load 2ml Standard Soln. in Water 2ml Standard Soln. in Water
Wash 2ml DI water 2ml DI water
Elute 1 2ml Methanol with 100 mM 2ml Acetonitrile with 10 mM
ammonium acetate ammonium acetate
Elute 2 2ml Methanol with 100 mM 2ml Acetonitrile with 10 mM
ammonium acetate ammonium acetate
Elute 3 2ml Methanol with 100 mM 2ml Acetonitrile with 10 mM
ammonium acetate ammonium acetate

Screening runs of the samples by GCMS showed significant degradation of the target
compounds preventing accurate quantitation. We therefore used LCMS to analyze the
starting, load, wash and elute solutions prepared by SPE.

The resulting LCMS chromatograms showed good retention of the compounds and revealed
that methanol with 100 mM ammonium acetate is an effective elution solvent. 94% or greater
recovery was observed for all three pesticides using methanol with 100 mM ammonium
acetate. Acetonitrile with 10 mM ammonium acetate also exhibited good results, with 90% or
greater recovery in the first elute step.

Individual Test Results

A summary of the individual test results is provided below. All accompanying data,
including spectra, has been included in the data section of this report.

SPE Sample Preparation

The three standards were added to 20 mL of water at a concentration of 0.5 mg/mL each.
The solution was thoroughly mixed on an orbital shaker. An Hydroclean RP-200mg
cartridge was prewashed with 2 mL of methanol and then 2 mL of water. 2 mL of sample
was then loaded onto the cartridge. The cartridge was then washed with 2 mL of water. The
first elution was performed using 3 successive aliquots of 2 mL of methanol with 100 mM
ammonium acetate. A second experiment was performed using the same load conditions
with 3 successive elute steps using 2 mL of acetonitrile with 10 mM ammonium acetate.
GCMS
Initial GCMS screening runs of the starting solution were analyzed at injection port
temperatures of 310 and 200°C. The resulting chromatograms showed significant
degradation of the target compounds as indicated by the number of peaks present. The
degree of degradation interfered with quantitation of these analytes using GCMS.

For analytes that are easily ionized, LCMS can provide improved detection because the
sample is not vaporized at high temperature prior to chromatographic separation. We
therefore continued the analysis using LCMS.

LCMS
The SPE starting, load, wash, and elute solutions were analyzed by LCMS in order to
determine their SPE retention behavior. The chromatogram of the SPE starting solution can
be found in Figure 1. The three standards were found to be adequately separated using a
gradient from 5% organic to 100% organic.

Aldicarb-
Sulfone
o-aminoazotoluene
Aldicarb
Oxamyl

Figure 1: LCMS chromatogram of the SPE starting solution identifying the three
standard peaks

The relative peak areas for each standard are provided in Table 1. Recovery values are
provided as well, and are based on the peak area relative to the starting solution.
 Table 1: Standard Peak Areas and % Recovery Values – Methanol SPE

Aldicarb Aldicarb Aldi-Sulf Aldi- Oxamyl Oxamyl

SPE Solution
Area % Area Sulf % Area %
Starting Solution 19820683 -- 27925760 -- 30229531 --
Load not detected 0.0 4088 0.0 861 0.0
Wash not detected 0.0 547619 2.0 556026 1.8
Methanol Elute 1 19394269 97.8 26322752 94.3 31301336 103.5
Methanol Elute 2 711011 3.6 784413 2.8 596237 2.0
Methanol Elute 3 164245 0.8 185503 0.7 201732 0.7

Table 2: Standard Peak Areas and % Recovery Values – ACN SPE

Aldicarb Aldicarb Aldi-Sulf Aldi- Oxamyl Oxamyl

SPE Solution
Area % Area Sulf % Area %
Starting Solution 19820683 -- 27925760 -- 30229531 --
Load not detected 0.0 97116 0.3 2455 0.0
Wash 1194 0.0 1762658 6.3 1107255 3.7
ACN Elute 1 18015467 90.9 25509864 91.3 29534958 97.7
ACN Elute 2 609237 3.1 714981 2.6 663020 2.2
ACN Elute 3 122213 0.6 243297 0.9 180476 0.6

Nearly 100% of the standards were retained on the loading step, and only a small percentage
was observed to elute with additional washing. A majority of the sample (≥ 90%) was
recovered during the first elute step.

For this analysis, approximately 40 ppm o-aminoazotoluene was added to each solution as an
internal standard. The peak areas in the above table are scaled based on the o-
aminoazotoluene response.

Analysis Conditions
This section of a Jordi report provides information on the methods used including
instrument type, temperatures, solvents, sample preparation, etc. The specific conditions
have been removed for this case study.

Closing Comments
Deformulation of an unknown material is intended to provide a best estimate of the chemical
nature of the sample. All chemical structures are supported by the evidence presented but are
subject to revision upon receipt of additional evidence. Additional factors such as material
processing conditions may also affect final material properties.

Jordi Labs’ reports are issued solely for the use of the clients to whom they are addressed. No
quotations from reports or use of the Jordi name is permitted except as authorized in writing.
The liability of Jordi Labs with respect to the services rendered shall be limited to the amount
of consideration paid for such services and do not include any consequential damages.

Jordi Labs specializes in polymer testing and has 30 years experience doing complete
polymer deformulations. We are one of the few labs in the country specialized in this type of
testing. We will work closely with you to help explain your test results and solve your
problem. We appreciate your business and are looking forward to speaking with you
concerning these results.

Sincerely,

Mark Jordi
Mark Jordi, Ph. D.
President
Jordi Labs LLC