EPA-R2-72-079 October 1972 Environmental Protection Technology Series Liquid Chromatography of Carbamate Pesticides National Environmental Research Center Office of Research and Monitoring U.S. Environmental Protection Agency Corvallis, Oregon 97330 ------- RESEARCH REPORTING SERIES Research reports of the Office of Research and Monitoring, Environmental Protection Agency, have been grouped into five series. These five broad categories were'established to facilitate further development and application of environmental technology. Elimination of traditional grouping was consciously planned to foster technology transfer and a maximum interface in related fields. The five series are; 1. Environmental Health Effects Research 2. Environmental Protection Technology 3. Ecological Research H. Environmental Monitoring 5. Socioeconomic Environmental Studies This report has been assigned to the ENVIRONMENTAL PROTECTION TECHNOLOGY series. This series describes research performed to develop and demonstrate instrumentation, equipment and methodology to repair or prevent environmental degradation from point and non-point sources of pollution. This work provides the new or improved technology required for the control and treatment of pollution sources to meet environmental quality standards.. ------- EPA-R2-72-079 October 1972 LIQUID CHROMATOGRAPHY OF CARBAMATE PESTICIDES by Alfred D. Thruston, Jr. Southeast Environmental Research Laboratory College Station Road Athens, Georgia 30601 Project 16020 EWC Program Element 1B1027 NATIONAL ENVIRONMENTAL RESEARCH CENTER OFFICE OF RESEARCH AND MONITORING U.S. ENVIRONMENTAL PROTECTION AGENCY CORVALLIS, OREGON 97330 For sale by the Superintendent of Documents, U.S. Government Printing Office Washington, B.C., 20402 - Price 55 cents ------- ABSTRACT A commercial liquid chromatograph was evaluated and found useful for analysis of carbamate pesticides. Liquid chromatography retention times for 23 carbamate pesticides are given. The UV detector required 20 to 1500 ng for the pesticides studied to give a 25% full- scale recorder response. 111 ------- CONTENTS Section Page I Conclusions 1 II Introduction 3 III Experimental 5 IV Results and Discussion 9 V References 15 v ------- FIGURES PAGE 1 CHROMATOGRAM OF CARBAMATE PESTICIDES ON PERMAPHASE ODS COLUMN 11 2 CHROMATOGRAM OF CARBAMATE PESTICIDES ON PERMAPHASE ETH COLUMN 12 3 CHROMATOGRAM OF SWEP, EXTRACTED FROM WATER SPIKED AT 2 yg/£ LEVEL 14 VI ------- TABLES No. Page 1 Chemical Names and Sources of the Carbamate Pesticides 6 2 Retention Times and Sensitivities of Various Carbamates on Permaphase ODS Column 10 3 Retention Times and Sensitivities of Various Carbamates on Permaphase ETH Column 13 VI1 ------- SECTION I CONCLUSIONS Liquid chromatography shows promise for analysis of carbamate pesticide residues. The technique provides separations with speed and resolution comparable to those of gas chromatography. Ambient or near ambient temperature can be used with heat-labile compounds. Sensitivity is 2 to 3 orders of magnitude less than that of electron capture gas chromatography but is still adequate for many compounds. ------- SECTION II INTRODUCTION Most carbamate pesticides cannot be analyzed directly by gas chromatography unless conversion to more suitable derivatives is carried out (1/2), because they are thermally unstable. Techniques, such as semi- quantitative thin-layer chromatography (3,4) or quan- titative ultra-violet spectrometry (5), vary in selectivity and sensitivity. Liquid chromatography (LC) is a relatively new analytical tool that offers selectivity and moderate sensitivity for analysis of these heat-labile compounds. LC separation and detection of Sevin (carbaryl) and its hydrolysis product, 1-naphthol, from plant extracts have been reported (6), but no comprehensive list of retention times and sensitivities of various carbamate pesticides has been published. A liquid chromatograph is similar to a gas chromatograph; each has an injection port, a packed column, and a detector. A liquid mobile phase under pressure is used in LC instead of a carrier gas as in gas chromatography. Many compounds that are not volatile enough or too heat- labile for analysis by gas chromatography will partition between the LC stationary and mobile phases. Two DuPont stationary phase columns were evaluated for analysis of carbamate pesticides—Permaphase ODS (for non-polar compounds) and Permaphase ETH (for more polar type carbamates). ------- SECTION III EXPERIMENTAL Liquid ahromatograph.--A DuPont Model 820 liquid chromatograph equipped with an ultraviolet photometric detector that measures the absorbance at 254 nm was used. The following columns and conditions were employed: (1) 1m x 2mm id stainless steel column, packed with Permaphase ODS (octadecyl silane). Mobile phases were 6% and 30% methanol in water. A pressure of 1000 psi at 50° C maintained a flow of 1 ml/min. (2) 1m x 2mm id stainless steel column, packed with Permaphase ETH (ether). Mobile phases were hexane, 1% isopropanol/hexane, and 4% isopropanol/hexane. A pressure of 400 psi at 40° C maintained a flow of 1 ml/min. Solvents.--Spectrograde hexane, isopropanol, methanol and methylene chloride. Standard carbamate solutions.--Standards (Table I) were prepared as isopropanol solutions. ------- Table 1. Chemical names and sources of the carbamate pesticides Common or Trade Name Chemical Name Source BaygonR (aprocarb) FuradanR (carbofuran) MatacilR (aminocarb) MobamR SevinR (carbaryl) Landrin UC 10854 UC 8454 Carbanolate RE 5305 Mesurol Zectran1 R 2-isopropoxyphenyl-N-methyl carbamate 7-(2 ,3-dihydro-2,2-dimethyl)- benzofuranyl-N-methyl carbamate 4-dimethylamino-3-methylphenyl- N-me'thyl carbamate 4-benzothienyl-N-methyl carbamate 1-naphthyl-N-methyl carbamate 3,4,5-trimethylphenylmethyl- carbamate 3-isopropylphenyl-N-methyl carbamate 1- (5,6,7,8-tetrahydro)-naphthyl- N-methyl carbamate 3,4-xylyl-6-chloro-N-methyl carbamate 3-sec-butylphenyl-N-methyl carbamate 4-methylthio-3,5-dimethylphenyl- N-methyl carbamate 4-dimethylamino-3,5-dimethy1- phenyl-N-methyl carbamate Chemagro Corp. Niagara Chem. Div., FMC Corp. Chemagro Corp. Mobil Chem. Co. Union Carbide Corp. Shell Development Corp. Union Carbide Corp. Union Carbide Corp. Upjohn Co. Chevron Chem. Co. Chemagro Corp. Dow Chem. Co. ------- Table 1. Chemical names and sources of the carbamate pesticides Common or Trade Name Chemical Name Source SirmateR, 2,3-isomer Sirmate/ 3,4-isomer BuxR AzakR (terbutol) Benomyl (Dupont Fungicide 1991) IPC CIPC DimetilanR TemikR Swep Barban 2/3-Dichlorobenzyl-N-methylcarbamate 3,4-Dichlorobenzy1-N-methylcarbamate 3-(l-Methylbutyl) phenyl methylcarbamate 2,6-Di-tert.-butyl-4-methylphenyl- N-methylcarbamate Methyl l-(butylcarbamoyl)-2- benzimidazole carbamate isopropyl-N-phenyl carbamate isopropyl-N-(3-chlorophenyl) carbamate 3-(l-N,N-dimethylcarbamoyl-5- methyl)-pyrazolyl-N,N-dimethyl carbamate 2-methyl-2-methylthio propion- aldehyde-0-methylcarbamoyl oxime methyl-N-(3,4-dichlorophenyl) carbamate 4-chloro-2-butynyl-N-(3-chloro- phenyl) carbamate FDAa FDAa Chevron Chem. Corp. Hercules Powder Co. E. I. DuPont de Nemours and Co. PPG PPG Geigy Chem. Corp. Union Carbide Corp. Niagara Chem. Div., FMC Corp. Gulf Res. & Dev. Co. ------- Table 1. Chemical names and sources of the carbamate pesticides Common or Trade Name Chemical Name Source Thiram bis(dimethylthiocarbamoyl) E.I. Dupont de Nemours disulfide and Co. MyloneR(dazomet) 3,5-dimethyl-l,3,5- Union Carbide Corp. tetrahydrothiadiazine-2-thione LannateR(methorny1) methyl N-[ (methy lcarbamoyl)oxy]- E.I. Dupont de Nemours thioacetimidate and Co. aFood and Drug Administration, Washington, D.C, 00 ------- SECTION IV RESULTS AND DISCUSSION Table 2 lists retention times and sensitivities for 17 carbamates on Permaphase ODS. The first 12 eluted as sharp symmetrical peaks within 10 minutes with 6% methanol in water as the mobile phase; Figure 1 shows a chromatogram of five of these carbamate pesticides and 1-naphthol. The last five in Table 2 had excessive retention times, which were shortened by changing the mobile phase to 30% methanol in water. Figure 2 shows chromatograms of seven of the more polar carbamates on the Permaphase ETH column. A change in mobile phase polarity from 1% isopropanol in hexane to 4% isopropanol in hexane causes significant changes in retention times. Table 3 summarizes retention times and minimum sensitivities of nine polar carbamates. For these carbamates, the sensitivity of the UV detector ranges from 20-1500 ng, a level suitable for pesticide residue analysis. This work was done with pure solutions to obtain standard retention times. Natural sample extracts were not studied systematically. The LC technique was applied to a study of the kinetics of Sevin degradation by micro-organisms. Aqueous samples of 4-8 yl were injected into the LC without prior clean- up or extraction. Sevin was observed in these samples in the 11-33 mg/A range based on comparison with standard solutions. Considerably lower detection limits can be obtained by concentrating the sample. For example, a liter of tap water, spiked with 2 yg/1 of swep (methyl dichlorophenyl carbamate) was extracted with methylene chloride (3) and concentrated to 200 yl. Figure 3 shows a distinct swep peak in the chromatogram of a 5-yl injection of the concentrate. ' ------- Table 2. Retention times and sensitivities of various carbamates on Permaphase ODS column, 50° C, methanol/ water mobile phase, 1000 psi,lml/min flow rate, UV detector. Chemical Name Baygon Furadan Matacil Mob am Sevin Landrin UC 10854 UC 8454 Carbanolate l-naphthola RE-5305 Mesurol Zectran Sinuate, 2,3- isomer Sirmate, 3,4- isomer Bux Benomyl Azak 6% MeOH/ H20 Rj.Cmin) 1.50 1.60 2.00 2.10 2.60 3.25 3.40 3.50 3.60 4.30 7.20 7.65 8.30 30% MeOH/ H20 Rt(min) 2.30 2.60 3.50 6.10 6.35 Min . Amount to give 25% FSD (ng) 250 300 50 25 100 1000 1000 1000 1000 250 1000 200 200 1500 1500 1500 1500 1500 ahydrolysis product of Sevin Abroad skewed peak ------- 4 Minutes 8 Figure 1. Chromatogram of carbamate pesticides on Perma- phase ODS column. 6% methanol/94% water mobile phase. 1, Furadan; 2, Matacil; 3, Sevin; 4, Landrin; 5, 1-naphthol (hydrolysis product of Sevin); 6, Mesurol. 11 ------- H to (a) (b) 4 Minutes 8 Figure 2 Chromatogram of carbamate pesticides on Permaphase ETH column: (a) 1% isopropanol/99% hexane mobile phase. 1, CIPC; 2, Dimetilan; 3, Temik; 4, barban. (b) 4% isopropanol/96% hexane mobile phase. 1, swep; 2, barban; 3, Mylone; 4, Lannate. ------- Table 3. Retention times and sensitivities of various carbamates on Permaphase ETH column, 40 °C, 0 to 4% isopropanol/hexane mobile phase, 400 psi,lml/min flow rate, UV detector. Chemical Name I PC CIPC Dimetilan Temik Swep Barb an Thiram Mylone Lannate Hexane Rt (min) 1.52 1.92 1% IPA/Hex R-t (min) 1.50 1.50 2.10 2.66 3.00 7.20 4% IPA/Hex R£ (min) 1.68 1.80 2.96 2.90 3.82 4.88 Min. Amount to give 25% FSD (ng) 250 500 1000 100 20 400 50 100 500 13 ------- V 0 J 0 4 Minutes Figure 3. Chromatogram of swep, extracted from water spiked at 2 yg/5. level. 1, solvent peak; 2, swep. 14 ------- SECTION V REFERENCES 1. Holden, E.R., Jones, W.M. , and Beroza, M. , "Determination of Residues of Methyl- and Dimethyl Insecticides by Gas Chromatography of Their 2,4-Dinitroaniline Derivatives," J. Agr. Food Chem., 17, No. 1, pp 56-59 (1969). 2. Butler, L.I. and McDonough, L.M., "Specific GLC Method for Determining Residues of Carbaryl by Electron Capture Detection after Derivative Formation," JAOAC 53, No. 3, pp 495-498 (1970). 3. Abbott, D.C., Blake, K.W., Tarrant, K.R., and Thomson, J. , "Thin-Layer Chromatographic Separa- tion, Identification, and Estimation of Residues of Some Carbamate and Allied Pesticides in Soil and Water," J. Chromatog. 30, pp 136-142 (1967). 4. El-Dib, M.A., "Thin-Layer Chromatographic Detection of Carbamate and Phenylurea Pesticide Residues in Natural Waters," JAOAC 53, No. 4, pp 756-760 (1970). 5. Aly, O.M., "Spectrophotometric Determination of Sevin in Natural Waters," JAWWA 59, No. 7, pp 906- 912 (1967) . 6. "The Analysis of Carbaryl and Its Major Hydrolysis Product 1-Naphthol in Plant Extracts," Chromato- graphic Methods 820 M7, February 13, 1970 (a DuPont Instruments" Information Sheet). 15 ------- 1 5 /4rcc.v.Mori Numbt-r 2 Subject Fu'ld & Group 05A SELECTED WATER RESOURCES ABSTRACTS INPUT TRANSACTION FORM orgamzat.on u>s> Environmental Protection Agency Southeast Environmental Research Laboratory Athens, Georgia Title LIQUID CHROMATOGRAPHY OF CARBAMATE PESTICIDES 10 Authors) Thruston, A.D., Jr. 16 Project Designation PROJECT # 16020 EWC 21 Note 22 Citation Environmental Protection Agency report number EPA-R2-72-OT9, October 1972. 23 Descriptors (Starred First) *Analytical techniques, *Chromatography, *Pesticides, *Carbamate pesticides. 25 Identifiers (Starred First) *Liquid chromatography, retention times. 27 Abstract A commercial liquid chromatograph was evaluated and found useful for analysis of carbamate pesticides. Liquid chromatography retention for 23 carbamate pesticides are given. The UV detector required 20 to 1500 ng for the pesticides studied to give a 25% full-scale recorder response. (Thruston—Southeast Environmental Research Laboratory) Abstractor Tbr-nat-rm -TV. Institution Southeast Environmental Research Laboratory WR;102 (REV. JULY 1969) WRS1 C SEND TO: WATER RESOURCES SC I EN Tl FIC INFORMATION CENTER U.S DEPARTMENT OF THE INTERIOR WASHINGTON. O. C. 20240 -. U. S. GOVERNMENT PRINTING OFFICE : 1972-514-149/74 « Sl>0: I9S9-SS9-339 ------- |