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LCMS-8040 Application Pesticides in Cannabis

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LCMS-8040 Application Pesticides in Cannabis LCMS-8040 Application Pesticides in Cannabis Je Dahl,1 Julie Kowalski,2 Jason Zitzer,3 and Gordon Fagras3 1Shimadzu Scientic Instruments; Columbia, Maryland 2Restek Corporation; Bellefonte, Pennsylvania 3Trace Analytics; Spokane, Washington Pesticides in Cannabis Summary: An LC-MS method for detection of fortification standards as appropriate. A multi- pesticides in cannabis with QuEChERS extraction residue pesticide mix was used (Restek part was developed. number 31971). Background: Medicinal and recreational use of After hydration of the samples, 15 mL acetonitrile cannabis has increased rapidly in recent years. with 1% acetic acid was added to each followed Like other crops, cannabis is susceptible to by shaking for 30 minutes. To each sample was insects, mold, and chemical residue contamina- added the contents of an AOAC QuEChERS tion. Pesticides and antifungals have been applied Packet (Restek part 26237) and the samples were to cannabis to increase yields however these vigorously mixed for 2 minutes and centrifuged. substances may cause human harm if they are consumed by users. Sensitive and selective Dispersive SPE was used to clean up the sample detection of these residues is necessary for con- extracts for LCMS analysis. Several formulations sumer protection. QuEChERS extraction and of cleanup reagents were tested for optimum LC-MS analysis offers effective and efficient combination of matrix removal and recovery. The detection of such chemical residues. best formulation was a combination of PSA, C18, Method: Cannabis samples were provided by licensed growers in Spokane, Washington, and samples were prepared and analyzed in a certified lab in that state. Pesticide-free organically-grown cannabis was used for spiking studies and calibra- tion curves. A variety of cannabis samples offered for retail sale were analyzed for pesticides as well. Cannabis dried flower samples were homogenized by an automated grinder and weighed portions of 1.5 grams were used for each analysis. Each ground sample was hydrated using 15 mL of 1% acetic acid in water and agitated for 30 minutes. Figure 1: Moldy cannabis flower. The white As can be seen, only a few compounds have mized and measured for pesticide residues. The Samples were spiked with internal standards and powder is mold growth which can flourish in recovery less than 50% or greater than 120%. results are shown in the table. The most com- high humidity growing operations. One notable exception is bifenazate, which monly detected pesticide was piperonyl butoxide, showed a strong matrix enhancement effect. This and was detected over a wide range of concentra- effect was confirmed by spiking experiments in a tions. Piperonyl butoxide finds wide use in pesti- variety of cannabis specimens and should be cide formulations to enhance activity of the main and carbon in the ratio 50:50:7.5 in addition to pole mass spectrometer. The LC column was a considired whenever a signal for bifenazate is ingredient. Myclobutanil, an antifungal known to MgSO4 (Restek part 26243). The supernatants Restek Ultra-AQ C18 column (3 µm, 2×100 mm) detected. be used in cannabis cultivation, was detected in a were added to the dispersive SPE reagents, and a binary gradient was used of 5 mM ammo- number of samples as well. mixed vigorously, and centrifuged. The superna- nium formate with 0.1% formic acid (pump A) and Limits of quantitation were determined by measur- tants were removed and concentrated approxi- methanol (pump B). The flow rate was 0.5 mL/min, ing samples in triplicate at various levels. Signal- Conclusion: An effective method for detection of mately 5-fold by evaporation prior to analysis. the column temperature was 40 °C, and the total to-noise of at least ten to one and RSD of 20% or pesticides in cannabis is demonstrated using Each sample was centrifuged to remove particu- run time was 15 min. A divert valve was used to better were required at the limit of quantitation. QuEChERS sample extraction with dispersive lates before being placed in an autosampler vial. send the unretained and re-equilibration portion of The limits of quantitation are reported in the table. SPE cleanup and LC-MS analysis with a triple the gradient to waste to reduce intstrument fouling quadrupole mass spectrometer. The method was LCMS analysis was carried out on a Shimadzu and the injection volume was limited to 1 µL. A variety of cannabis samples offered for retail applied to detect a number of pesticides in comer- Prominence HPLC with LCMS-8040 triple quadru- sale and presented to us for analysis were anony- cially available cannabis samples. Figure 2: Representative dried cannabis flower samples (anonymized) tested for pesticides. Page 2 Electrospray ionization in continuous polarity with good peak shape and retention time repro- switching mode was used for detection. Optimized ducibility. Individual chromatograms from selected MRM settings were used for each compound and pesticides are shown in the figure. The calibration at least one quantifier and one qualifier transition curve was prepared in spiked matrix over the was selected. The retention times were found and range of 3.91 to 500 ppb, as dried flower weight. used to program the MRM segments for optimum Individual calibration curves for selected com- duty cycle. pounds are shown as well. Results and Discussion: A representative chro- The total recovery for each compound, as mea- matogram of a prepared, spiked cannabis sample sured by comparing the peak area for a 50 ppb is shown in the figure. The total run time was 15 spiked sample with a spiked process blank, which min. and chromatographic peaks were observed represents 100% recovery and no matrix effects. Summary: An LC-MS method for detection of fortification standards as appropriate. A multi- pesticides in cannabis with QuEChERS extraction residue pesticide mix was used (Restek part was developed. number 31971). Background: Medicinal and recreational use of After hydration of the samples, 15 mL acetonitrile cannabis has increased rapidly in recent years. with 1% acetic acid was added to each followed Like other crops, cannabis is susceptible to by shaking for 30 minutes. To each sample was insects, mold, and chemical residue contamina- added the contents of an AOAC QuEChERS tion. Pesticides and antifungals have been applied Packet (Restek part 26237) and the samples were to cannabis to increase yields however these vigorously mixed for 2 minutes and centrifuged. substances may cause human harm if they are consumed by users. Sensitive and selective Dispersive SPE was used to clean up the sample detection of these residues is necessary for con- extracts for LCMS analysis. Several formulations sumer protection. QuEChERS extraction and of cleanup reagents were tested for optimum LC-MS analysis offers effective and efficient combination of matrix removal and recovery. The detection of such chemical residues. best formulation was a combination of PSA, C18, Method: Cannabis samples were provided by licensed growers in Spokane, Washington, and samples were prepared and analyzed in a certified lab in that state. Pesticide-free organically-grown cannabis was used for spiking studies and calibra- tion curves. A variety of cannabis samples offered for retail sale were analyzed for pesticides as well. Cannabis dried flower samples were homogenized by an automated grinder and weighed portions of 1.5 grams were used for each analysis. Each ground sample was hydrated using 15 mL of 1% acetic acid in water and agitated for 30 minutes. As can be seen, only a few compounds have mized and measured for pesticide residues. The Samples were spiked with internal standards and recovery less than 50% or greater than 120%. results are shown in the table. The most com- One notable exception is bifenazate, which monly detected pesticide was piperonyl butoxide, showed a strong matrix enhancement effect. This and was detected over a wide range of concentra- effect was confirmed by spiking experiments in a tions. Piperonyl butoxide finds wide use in pesti- variety of cannabis specimens and should be cide formulations to enhance activity of the main and carbon in the ratio 50:50:7.5 in addition to pole mass spectrometer. The LC column was a considired whenever a signal for bifenazate is ingredient. Myclobutanil, an antifungal known to MgSO4 (Restek part 26243). The supernatants Restek Ultra-AQ C18 column (3 µm, 2×100 mm) detected. be used in cannabis cultivation, was detected in a were added to the dispersive SPE reagents, and a binary gradient was used of 5 mM ammo- number of samples as well. mixed vigorously, and centrifuged. The superna- nium formate with 0.1% formic acid (pump A) and Limits of quantitation were determined by measur- tants were removed and concentrated approxi- methanol (pump B). The flow rate was 0.5 mL/min, ing samples in triplicate at various levels. Signal- Conclusion: An effective method for detection of mately 5-fold by evaporation prior to analysis. the column temperature was 40 °C, and the total to-noise of at least ten to one and RSD of 20% or pesticides in cannabis is demonstrated using Each sample was centrifuged to remove particu- run time was 15 min. A divert valve was used to better were required at the limit of quantitation. QuEChERS sample extraction with dispersive lates before being placed in an autosampler vial. send the unretained and re-equilibration portion of The limits of quantitation are reported in the table. SPE cleanup and LC-MS analysis with a triple the gradient to waste to reduce intstrument fouling quadrupole mass spectrometer. The method was LCMS analysis was carried out on a Shimadzu and the injection volume was limited to 1 µL. A variety of cannabis samples offered for retail applied to detect a number of pesticides in comer- Prominence HPLC with LCMS-8040 triple quadru- sale and presented to us for analysis were anony- cially available cannabis samples.
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