Cover Sheet for ENVIRONMENTAL CHEMISTRY METHOD Pestcide Name: Metaldehyde MRID #: 422870-01 Matrix: Si Analysis: GC/NPD ‘This method is provided to you by the Environmental Protection Agency's (EPA) Environmental Chemistry Laboratory (ECL). This method is not an EPA method but one, which was submitted to EPA by the pesticide manufacturer to support product registration, EPA recognizes that the methods may be of some utility to state, tribal, and local authorities, bbut makes no claim of validity by posting these methods. Although the Agency reviews all * Environmental Chemistry Methods submitted in support of pesticide registration, the ECL evaluates only about 30% of the currently available methods. Most methods perform satisfactorily but some, particularly the older methods, have deficiencies. Moreover, the print quality of the methods varies considerably because the methods originate from different sources. Therefore, the methods offered represent the best available copies. If you have difficulties in downloading the method, or further questions concerning, the methods, you may contact Elizabeth Flynt at 228-688-2410 or via e-mail at ov.i 4, 90-0035 Lo Page 118 S$ ANALTTICAL 2389 feringion Point Ox. © Weston Salem, PC. 27107 te Phone (919) 785-3252.3 EN-CAS Method No. ENC-2/91. Phone: ‘ (919) 785-3252 ‘Total Pages = 43EN-CAS Project # 90-0035 Lo EN-CAS Method No. ENC-2/91 WOME PAGE © ee ee ee 2.0 3.0 4.0 s.0 T0 TEST SUBSTANCES... =... 2... e PREPARATION OF ANALYTICAL STANDARDS. . 5:2 Gas 5.1 Fortification Standards... . ‘Chromatographic Standards condit: Sie Calibration © 22 DD Gc Analyses of Samples. | Representative Curomatograns. Description and Typical operating Page 119 Page 2 ee eS ew ee ee 20 20 an 1 12 a2 3 Fry ueEN-CAS Project # 90-0035 Lo Page 120 { e ireas rated Ho. BI6-2/81 rage 3 TABLE oF ComTErrs (continued) 8.0 CALCULATIONS © 2. ee ee ee 2 calculation of ng Found + + - \calation of ag-Bquivaiests i Canculation of Soll Moisture 7.02. 2 Las Calculation of Soll Residues of dey Heigit a Calculation of Percent Recovery > 222222 las : Beample Calculation... --- 2-22-22 219 I 9.0 VALIDATION RESULTS... 1... eee ee 20 | 9.1 Statistical Method. ©. ee ee 20 | 9:2 Discussion of Validation Results. > ° 2: 2! 20 r ) 10.0 REFERENCES © 2 2 ee ee et 22.0 TABLE Table I Metaldehyde in San Joaguin | California Soll... se... 1. 2 22 12.0 FIGURES i Figure 2. Fiowchart of Method ENC-2/91. . . . 25 Figures 2-14 Typical chromatography... .. 1 26 Figures 15 - 16 Typical calibration curves. |: ! ! 39 13.0 APPENDIX A. EN-CAS SOP ITI-5.9 entitled Processing soil Sora Samples then Application Ia By Keans of dasued 11/02/91. - 0 ee ee et ee ADEN-CAS Project # 90-0035 Lo Page 121 e ics aecsed Ho, m1e-2/21 rere 6 ORE Analytical method for the Deteraination of ataldebode asd 2.0 ermonocrion 2.1 Scope ‘This nethod is used for the determination of Bigot Material (Reference 3) was used in this uethod. ‘The limit of quantitation (109) is 0.02 ppm (g/g) matsidehyde. Method validation Fesalts from EN-CAS report 90-0035 Lo Daterainaticn jtre included in ehis report (see Table I). See Figure 1 for a flowchart of the mathed. 1.2 Principle Metaldehyde is extracted from soil by shaking with dichloromethane (DCM) that has been pre-vashed with 2% aqueous sodium bisulfite to remove traces of the contaminant, acetaldehyde. An aliquot of the DON oxcract is washed vith 2% aquecus sodium bisulfite to Fenove extracted aldehydes which may cause CC interferences after derivatization. After i the DCM phase is cerivatized by shaking with a 0.5¢ ution of 2,4~dinitrophenylhydrazine (2,4-DaP) in r'to use, the derivatizazion reagent is pre-vashed with DCM to remove traces of acetaldehyde BEST AVAILABLE COPYEN-CAS Project # 90-0035 Lo Page 122 @ BI-CAS Mathod No. ENC~2/92 rage 3 1.2 Principle (continued) 2,4-OMPH which may be present as a contaminant). Under the acidic derivatization conditions, tha metaldehyde is converted to acetal: which then dissolves ee eee with the 2, acataldebyde 2,4-DXP=. The dehyde 2, soluble and is a therefore retained £2 the Dot phase as it is formed : during tha darivacizacion-shaking Process. Tue Feaction aixeure is transferred o's separatory Funnel and the organic phase contaizing the derivative is drained beck into the sesction vessel (the aqueous phase ts discarded). “The organic passe ia returned $5 the separatory funnel, washed with ¢¥ Seaple {a then filtered throoye a ped of Sanparca’™ senple Ziltered a pad of sonyarous Sodium sulface and retuced to'=5 ML oy vacuus roCary evaporation at a bath taaparature cf 30°C 35. TS Fanove the last traces of Dom, etby: acetaze is added to the sample and the sizple 23 evaporated to dryness using vacuum Totary evaporaccon wich «bath feuperature cf 30°C ~ 35%. The sanrie extract i= r } adjusted to an appropriate final vormse. cas chromatographic jon is accomplished with a Eapillary DB-17 column. analyte dececcion ia obtained using an alkai{ flame nitrogen/phesphorus (1/2) detector. All equipment and apparatus aay be replaced by equivalent items from alternate sources. WOTE: Pricr to use, all labware should be rinsed with ich has been’ pre-washed with sodi== bisulfite). icetone ner methancl should be used to rinse Glassware. Acetone reacts with the 2,4-2¥P derivatizing agent giving rise to a potential interfe=erce. Methanol, contains small amounts of acetaldehyde weich can also interfere in the metaldehyde analyses. 2.1 French square bottles, 4-0z and 32-<= 2.2 Caps, polyethylene-lized, for French square bottles 2.3 Separatery tunnels, 7S2-aLEN-CAS Project # 90-0035 Lo Page 123 EB-CAS Method No. ENC-2/92 Page 6 2-0 ABPARATUS (continues) 2.4 Erlenmeyer flasks, 250-mL, with 24/40 ground glass fittings 2.24 Mettler analytical 2.5 Stoppers, ground glass, 24/40 2.6 Volumetric flasks, 100 mL, for preparing analytical standards 2.7 Power funnels, glass, 4-inch diaseter 2.3 Disposable Pasteur pipettes, 23-ca 2.9 Seintillation vials, 20-ar 2.20 Ge vials, 2-ab 2.21 Mechanical Shaker (G29 cyrotory) 2-22 Rotary Zvaporator, ccld fingers (Buchi Rotowapor, model #RE1i1) 2.23 Top loading balance, (Tisher Scientific, Model XT=3KD) balance capable of 5 decinel accuracy, for weighing analytical standards + Dichloromethane (Dan, pesticide grade. NOTE: Just prior to use (on the same day), wash the Dom with 2% (w+) sociun Bisulfite ealution. mis is accomplishe¢ by partitioning the DOM wits 5 parts 2% aqueous sodizm bisalfite to'1 part DCM. 3.2 Ethyl acetate, pesticide grade 3.2 Methanol, Fisher scientific, optina grade NOTE: All graces of nethanol are contaminated with spall amounts cz a The purest grade of zethanol available should be used. Hydrochloric acid, ACS Reagent grade Derivatizing reagent, 0.5% solution of 2,4— dinitzcphenylrerazize in 6N HCl (prepared daily)EN-CAS Project # 90-0035 Lo Page 124 @ ecas wethod No. E¥e-2/21 Page 7 Prepare derivatizing reagent by veighing 0.5 g of 2,4-dinit Anydrazine per 100 aL of GN HC1 (diiuted from'12N BCl). Warm, if necessary, to dissolve. Wash the reagent with 24 times its volume of pre—washed DoH on the day of use- 3.6 Sodium sulfate (Ne,S0.), anhydrous, ACS certified, heated in a muffle fursace at 6007 for 2 bours 3.7 Sodium bisulfite, ACS certified 3.8 B& J bottled water, HPLC grade 4.0 mest smmsrances: Cy OF @ Ped (Clig-C-0-CH-CBy i I o 0 r Pate RH (CHy-CE-0-CH-CBg Non 102 Metaldehyde Acetaldehyde 2,4- (CHa) ini: MW = 176.2 (2,4-DNPH) MW = 224.2EN-CAS Project # 90-0035 Lo Paga 125 e@ BH-CAS Hathod Ho. ENCH2/92 Page 8 4.0 ZEST SUBSTANCES (continued) Cg g0,-H*s 4 caycHo metaldendye > acetaldehyde CtigcH0 = = => cezgct#=N-NE-Celg (NOzto e scatalastyde —————»> acetaidenyde 2,4-na70 . 5.0 PREPARATION OF ANALYTICAL STANDARDS 5.1 Fortification standards Wala 100 mg (octive ingredient) oS, substance, metaldehyde, using an anal; balance into 8 106-aL volusetric flask. Dissolve and dilute to volume with methancl to prepare a 1000 pg/mL stock solution. (NOTR: All grades cf methapal are contaminated with svall amounts of acetaldehyde. i Use a grade vnich minimizes the acetaldehyde concentration. Fisher Optima grade specifies a maximum of 0.001% acctaldehyde, although in car qperience the amount present in typical lots appears to be significantly less than this.} Serially dilute the 1000 ug/ml stock solution in ethanol to prepare 100 ug/l and 10 po/al standard solutions. Use these solutions to fortity sail control samples in order to moniter procedural recovery. The stock standard solution is stable for approxinately 6 months. [ROTR: Store all standard stock solutions in a freezer at a temperature of wto’c to -17°C.}EN-CAS Project # 90-0035 LO Page 126 EN-CAS Method No. ENC-2/92 Page 9 5.2 Gas Chromatographic Standards derivatizing procedure. In this way, reagent in the standard is sufficiently similar to that in the samples to t direct 6c comparison of standards and samples. Note that experiments in our laboratory indicate that the tory following volumes of the 1 pg/aL metal solution: 0.1 mL, 0.2 aL, 0.5 mL andi mL. Hicher concentrations of standards are prepared directly from the 1000 pg/al metaldehyde stock solution in methanol and the following volumes added: 0.01 mL, 9,025 (aby 0-08 aly 0-1 at end 9-2 mb. mhan carried through the derivatizing procedure and recovered 10 mL of ethyl acetate, these standards will contain icetaldehyde 2,4-DNPH equivalent to the following metaldenyde concentrations: 0.01 pg/mL, 0-02 pg/mL, 0.05 ug/E, 0.10 pg/ml, 1.0 ug/aL, 2.5 pga, 5.0 wg/al,'10 ug/ml and 20 pg/aL.” Fresh standards are made at least weekly. (NOTE: Store all standart solutions in a refrigerator at a tesperatare of 1°c to 3°C.]EN-CAS Project # 90-0035 LO Page 127 e -CAS Hethod Ho. ENC=2/21 rage 10 6.0 ANALYTICAL PROCEDURES 6.1 Extraction 6.2.2 6.2.3 6.2.4 6.2.5 Pre-vash the dichloromethane by partii tioning with 5 2% aqueous sodium bisulfite to 1 part parts Discard the 2% aqueous sodiua Weigh 100 ¢ of soil (processed according to N-CAS sop’ ITT: -9, one Appendix a) into & bivoz French aquare Dotter” Make appropriate laboratary fortifications with the metaldenyde Stock solution prepared in Section S.1. Mininus volumes (usually less than or equal 1 aL) of the methanol fortification soluticn are used in order to minimize potential interferences from the traces of acetaldehyde in the methanol. Allow solvent from the fortification to evapcrate for approximately 20-30 minutes. Add 250 mL 'of DCM (pre-vashed with 2% aquecus sodium bisulfite; see MOTE above) to the French square bottle. Add 500 g of anhydrous sodiua salfate to the DoM/soil mixture. the bottle using a cap with rlene and place Le SS Lieu fee Place the bottle on its side in the mechanical shaker and shake vigorously (epprurimately 200 rpa) for 25 ninutes on one s! Duttle to'the opposite side and continue’ shaking for an additional 15 minutes. Remove the bottle from the shaker, place it upright and let the sample settle for at least 15 minutes. Partition and Derivatization 6.262 6.2.2 Transfer a 25~aL sample aliquot to a 250-L separatory funnel. Wash the DCM aliquot by shaking with 10 at cf 2% aqueous sodium bisulfite solution. allow the phases to separate. (NOTE: The aetaldehyde residues will remain in the erganic phase. ]EN-CAS' Project # 90-0035 LO Page 128 e EN-CAS Method No. ENC-2/91 Page 11 6.3 6.2.3 6.2.5 6.2.6 6.2.7 2.8 2 Partition and Derivatization (continued) Drain the organic phase into a 4-027 French square bottle. Discard the resaining aqueous wash. Add 10 mL of derivatizing reagent (0.5% solution of 2,4-DEP in 6N HCl - see Section 3.5). MOTE: The derivatizing reagent wust be washed th DCM (pre-washed with 2% aquecus sodium bisulfite) prior to use in order to remove be present. This is by Partitioning the derivecising agent with 2.5 tines its volume of pre-vashed OcH (see Section 6.1), then discarding the DCM. The derivatizing reagent is prepared daily. Cap the bottle securely using a cap with a polyathylene liner and mechanically shake (-200 rpm) the sample for 20 minutes. ‘Transfer the reaction aixture back to the 250-mL separatory funnel. (NOTE: Phase will contain the dinitropbenylnydrazone derivative.) Drain the organic phase back into the reaction vessel (the 4-oz French square bottle) and Giscara tha aquecus phase- Return the organic phase to the separatory fennel. wash the organic Sien'2'x bo'ax of aucl. "biseard both acid Wash the organic phase again by shaking with 2'x 20 aL of HPLC grade water.” Discard Doch water washes. Filtration and Evaporation 6.3.8 Filter the sample through a pad (approxizatel; 50 g) of anhydrous sodium suifate into a 250— lenmeyer flask with a ground glass neck. Einse the sodiun sulfate pad with 15 ai of Dot (pre-washed vith 2% aqueous sodiua bisulfite), combining the rinse with the filtrate.EN-CAS Project # 90-0035 Lo Page 129 @ BI-OAS mated Ho. 1IC-2/91 rage 12 6.3 6.4 viltration and Evaporation (continued) 6.3.2 Evaporate the sample to approximately 5 aL under vacuum ‘evaporation using a bath rotary, temperature of 30°C = 35°C. 6.3.3 add 25 mL of ethyl acetate and evaporate to 6.3.4 Dissolve the sample extract in an final volume (usually 10 aL) with ethyl acetate. Proceed to gas chromatographic analysis. Gas Chromatographic Determinaticn Use a 30 mx 0.32 mm, 0.25-um film thickness, capillary DB-17 column to achieve gas chromatographic separation. Use a Hevlett-Packart Model 5890-A-Cas Chromatograph (or equivalent) with an alkali flame, ni phorus (¥/P) detector For the soil types validated herein, this satnod is proven affective to a Log of 0.02 ppm metaldehyde. Adjust the instrument sensitivity, GC calibration standards and final sample volumes to allow detection of metaldehyda at 50% of the Log.EM-CAS Project # 90-0035 10 Page 130 e ees a 6.7 ‘Time Required for Analysis . An experienced technician can process a sat of approximately 12 samples (including controls and recoveries), and prepare then for injection on the ges chromatograph in approximately one 8-hour day. An additional day is required for reinjections and for annotating and calculating the data. Interference and Potential Probless 6.8 he derivatizing reagent (0.5% solution of 2,4~dinitrophenylhydrazine) is pre-washed with Shromatoarashic Interferences After the analyte bas eluted from the Gc, the column temperature is raised to 275% to burn off any long retention time interferences before the next injection.EN-CAS Project # 90-0035 Lo Page 132 @ BES wathod Ho. HIC-2/21 rage 14 The t which remains in the derivatized sample at the tine of a Chromatogr phy analysis has an enhanc! effect on the acetaldehyde 2,4-DNPH signal during capillary GC-"/P analysis. or accurate Deterioration of chromatography and baseline instability say be seen after as fev as 5° injections. When this is observed, the following maintenance functions are perforacd: ‘the inlet liner is replaced, the entirn injection port is cleaned (and the seals Feplaced), and approximately 0.5 to 1 mater of the front portion of the colum is removed. If these naintenance activities do net improve ‘the chrosatography, then the N/P detector bead should be replaced. 7.0 GAS CHROMATOGRAPHIC ANALYSIS. 7-1 Description and Typical operating Conditions Instrument: Hewlett-Packard Model 5890-A Gas Chromatograph with an alkali flame, nitrogen/phesphorus’ (N/P) detector equipped with a 7673A Automatic Sampler. Data is collected and Processed with a Hewlet=-Packard 3396A Integrator. column: DB-17, 30 mx 0.32 mm (3 6H Scientific), 0.25-ym film thickness Carrier: ge 4.80 at/min. Detector: 3, ale ‘Aux He 25.5 al/ain.EN-CAS Method No. EN-CAS Project # 90-0035 Lo ENC=2/92, 7.1 Description and Typical operating Conditions (continued) ; 1 aL, splitiess Program Initial oven Temperature = sovc initial Tine 1.5 min. Ramp A 4cyain. oven Teaperature 225°¢ tie = 7.9 nin. Bamp B = do*ain- Oven Temperature = 275°C Final Tine = 10 ain. ‘Typical Retantion ‘Tine: Ysomer #1 =~ 11.0 min. ‘ Tsomer #2 = ~ 11:3 min. 3 . 6. SURE AnD Ones TRESHELD ‘maa ors 7. SRN OFF REEORTIN OE LABELTG ‘TNR 0.000 TRG] + 8 8. TORS O8 STAPT/STOP MES : Mm oom DI + 29. Tmo Oma OE 0.000 DG} = 9 10, DKK oes OE ws ase +20 Ovo mam Yom io.so DA} «4 12> cu muse pms rm +2 1b. gH, ‘TMG 10.500 ary = 1. ST Pk SN PO ‘TE 1.70 Se Page 132 Page 15EN-CAS Project # 90-0035 LO Page 133 e ‘REEAS Hathod Ho. ENC-2/51 rege 16 7.2 calibration In order to bracket a wide range of etaldehyde” concentrations, two standard curves are used for quantitation. ‘Assuming a 10 mL final extract volume, the lower range curve (0.01 to 0.10 pq/ak) covers metals ‘soil concentrations from 0.02 to 0.1 ppm. The higher range curve (0.1 pg/ml to 20 B3/aL) Covers netaldehyde soil concentrations between 0.1 and approxizately 20 ppa. The demonstrated linear response range for is from 0.01 pg/ml to 20 pg/sl on the BP /P detector. le residue values and fortification values ‘the 0.2 pg/ml standard must be injected vithin the higher range standard curve (0-1 g/al to 20 yg/al). Tala approach allows reagent ‘samples and standards to remain constant. Only in rare cases would netaldenyde residues in the soil exceed 20 ppm after normal application. Use the metaldehyde Gc calibration standards that were prepared in the method (Section $.2) in concentrati ions ranging frem 0.01 yg/al to 0.10 pg/al or from 0.10 pg/aL to 20 pgyal to calibrate the instrument. Inject appropriate standards at the ‘of the run, after approxinately every 2 or 3 samples throughout the run, and at the end of the Fun. A linear regression function is generated using the resulting sumation peak height of the two isomers, see Section 8.0, (obtained from the integrator after subtraction of the reagent blank contribution) vs. nanograms injected. The correlation coefficient for the line should generally be equal to or greater than 0.990. The Sample nanograms found are detarained by inserting ‘the summation peak height value (corrected for the reagent blank) of the tvo isomers into the standard curve Linear regression equation. Gc Analyses of Samples The peak heights of the calibration standards are corrected for the concurrent reagent blank. Additionally, the peak heights of the control and treated samples are each corrected for the concurrent reagent blank. Any remaining control contribution is subtracted from the fortified recovery sample results. No correction for control is nade for treated residue samples.EN-CAS Project # 90-0035 Lo Page 134 e seas Mathod Ho. mHC=2/9 page i8 8.3 Calculation of ppm Found 8.4 Calculation of Soil Moisture Deteraine the percent of moisture in the soil by weighing 10 grass of soil in duplicate, drying gvernight at an oven temperature of 110, ‘and reveighing the soil. average ‘Guplica soll moistures is to be used in the residue calculation. Percent moisture in a soil sample: 8.5. Calculation of Soil Residues on Dry Weight Basis 8.6 Calculation of Percent Recovery When calculating percent recovery of a laboratory fortification, use the following ppm subtraction calculation if there is control contribution present.EN-CAS Project # 90-0035 Lo Page 135 WI-CAS Mathod Bo. ENC-2/91 Page 19 6 Calculation of Percent cxcery (emtiomad) yu fom (caractad) fectitieatice Led (rpm) 8 tecowery = 8.7 Example Calculation Temple: See B12, ren STE DOS Sela De EOS 1 same comes + (60 cmp comms 106 bart Aan ts = 483 cats 248 coms = (40.8 ext) ‘ tot = + 08 12.9 ots myemir, mgr Brod: Omg ipod = Burwerierim iat 210 my epi. pe (oorrecta) = 0.0262 ppm (sample) 9.0255 ppm (cocrnctd) 2 tacowery = rmeun 0.02 pp aedEN-CAS Project # 30-0035 Lo Page 136 e Ex-CAS Method No. BIC-2/92 page 20 9.0 VALIDATION RESULTS See Table I in this report. 9.1 Statistical method : ‘The mean recoveries and standard deviations are calculated from the validation data and appear in ‘the validation tables included in this method. ‘The standard deviation is calculated using the following equation: where the sum of the squares of the individual Suedteiata oe Set es e ips’ than the total mabe, "be weastrenuee trae Baty neil (oben Soa total nusbes oT neeeeeaenes ba its Bee 9.2 Discussion of Validation Results Recovery means and standard deviations calculated for the method validation in 0-6", 6-12" and 12-18" California soil indicace a reliable method for the determination of metaldenyde as 2,4-DNPH by gas chromatography. In spite of the extensive provisions in the xethod for nininizing backgromad from ambient acetaldehyde, a small but measurable acetaldehyde background appears in the reagent Slank. Small variations in this blank sonetines result in the appearance of somewhat elevated recoveries (~150%) at the sethod ra.EN-CAS Project # 90-0035 Lo Page 237 ER-CAS Method Wo. ENC-2/91 Page 21 10.0 REFERENCES eee eee eee eee 1) (4). 2. Environmental Protection Agency, August 17, uv. 1989, Pesticide 7 Good Laboratory Practice Federal Register Vol. 54: 34052-34074. 3. Research and Consulting Company AG-Mattod entitled Determination of Retaldehvde Residues in Plant Material.WI-CAS Project # 90~0035 LO Page 138 © Dreis met Ho. mH6-2/0 men 22 Pe (o/s) Fond mata Racoroed oe gi 8 €E SESEEE sed 3 i 393322 : ify a3 aj i Dt ae 53 = Bo afl ote rretee i i ! : z | 3 3 Z ” saaple splted during processing. aun eso gaaaaa .Page 139 WI-CAS Method No. miC-2/91 EN-CAS Project # 90-0035 LO “ayenieaynte oplgepreyan et peysodes pov evcnesplationgtaayyeyy-4't ee perseiea telecon 4 wheel ay eno on wheeler my eam welt a9 mame tweet 2 eet wheel td ete OM wiselan aly e006 ™. set erent sfselon tlt = 1 Awfselon a 2 Ce sHeuTeL a on 18 (utt-9) * Banged -uoTaEPT TEA pouIOH THB BTUI0ZF,TeD UTNbEOE Ue UT epAYSPTEIOEN (penuyquon) 7 givePage 140 EN-CAS Project # 90-0035 LO Page 24 Ewc-2/92 ‘EE-CAS Method Ho. “rua tern a pote pe secrete ye me a mM a wm oH ca a cm ts-tcaten tm ose tm. amen om oo te to Giat-2t) Bainseu UOTIEPT TEA PouIEH TYos eyuz0ZF TED UTNBeoe UNE UT EPAYOPTEION, (penuyqueo) x gTavsEN-CAS Project # 90-0035 Lo HE-CAS Mathod No. ENC-2/91 Page 241 Page 25 Extract setal by DCM (pre-vashed vith 2t equecus ‘sodium bisulfite) and solid anhydrous sodium sulfate to ‘the soil sample and mechanically shaking. 1 Derivatize an aliquot of the DCM extract by sechanically shaking with 2,4-dinitrophenylhydrazine in 6m BC] (pre—wasbed with DOM) 1 ‘Transfer the reaction mixture to «separatory funnal and (etter discarding the aqueous phase) wash the organic phase with ecid (discard acid washes). { Wash the organic phase with water (discard water washes). 1 Filter the sample through anhydrous sodius sulfate. 1 Evaporate the sample to a small volume using rotary evaporation. 7 1 Add ethyl acetate and evaporate to dryness. ' Reconstituta the residue with ethyl acetate to an appropriate ‘final volume. Analyze on a gas chromatograph using 4/P detection and a capillary DB-17 column. _EN-CAS Project # 90-0035 LO Page 142 e e-cas athod Ho. 2/91 age 26 Reagent Blank attenuation = 2 2 mew ure need tetas, Mataldehyde Bquivalents Found: <0.02 ppm (0.0061 ppm) GC run # 37243, sat MV-1, dated 11/17/91 @ * Expressed as setaldehyde equivalentsEN-CAS Project # 90-0035 LO Page 143 EN-CAS Method Ho. ENC-2/91 Page 27 FISuRE 3 ‘Typical _Corosatoccan Gc standard 01 pg/ak Acetaldehyde 2,4~Dinitropheny} ‘ (2,4-DNPE) as Metaldehyde Equivalents Attemation = 2* mum te-ma7@— loomer 1 scetaldehyte 24-20PE 2.120—@—toomer 2 acetaldehyde 24-200 0.01 ng injected Gc ran # 37243, set MV-1, dated 11/17/91 @ * 2xpressed as xecaldehyde equivalentsEN-CAS Project # 90-0035 Lo Page 144 e DecAS athe Ho. ZC+2/91 rage 24 ric « ‘Typical _chrumatooraa GC Standard 0.05 ug/mt. acetaldehyde 2,4-Dinitrophenylbydrazone (2,4-DNPH) as Metaidenyde Equivalents Attenuation = 2 t99e anes aq 1. 8zee—isomer 1 ecetakdehyde 24-00 anata omer 2 acetaldehyde 2¢-DRPE 9.05 ng injected Gc run § 37263, set MV=1, dated 11/18/91 e * Expressed ac metaldehyde equivalentsEN-CAS Project # 90-0035 LO Page 145 e@ T-CAS Method if. m16-2/91 Pepe 29 ‘FICURE 5 ‘Typical chresatocran Gc Standara, 1.0 ugyal Acataldehyde 2,4-Dinit (2,4-DNPH) as Metaidehyde Equivalents Attemmation = 2° suns 17 mow an ayn eaeazeae 1.0 ng injected Gc run # 27243, set MV-1, dated 12/18/92 e * 2xpressed as netalderyde equivalentsEN-CAS Project # 90-0035 Lo Page 146 e EN-CAS Method Ho. ENC-2/91 Page 30 Soil Control (o-s") Attenuation = 2° mune 7 mov a7. t99 ate G3321¢— tomer 1 acetaMakple 24-DmPE @ 12 -41¢¢—Ieomer 2 ecetaHehyde 24-DNPE* eco ZN-CAS Sample ID #: ET1a89-c2 Metaldehyde Zquivalents Found: <0.02 ppm (0.0041 ppm) GC run # 37243, set Mv=1, dated 11/17/91 e * Expressed as zetaldehyde equivalentsBN-CAS Project # 90-0035 LO. Page, 147 MN-CAS Method No. ENC-2/91 Page 32 FrcuRE 7 ‘Typical chromatooran ‘Soil Control + 0.02 ppm Mataldehyde 0-6") Low Fortification attenuation = 2° rye eae wow a7. ieee aarzanes steer EN-CAS Sample ID # Metaldehyde Equivalents Recovered: 131% zr1ss9-s2 Ge run # 37243, set MV-1, dated 11/17/51 a * Bipressed as netaldehyde equivalentsEN-CAS Project # 90-0035 Lo Page 148 ® feos mtd me. E91 tae FIGURE 8 ‘Typical chromatogras Soil Control + 1.0 ppm Mataldebyde 0-6") igh Fortification Attemation = 2° sue ae may te. L998 osteeras steer Ate113e—leomer 2 seats dehyde tere EN-CAS Sample ID #: E71889-s5 Metaldehyde Equivalents Recovered: 95% Gc run #37243, set MV-1, dated 11/18/91 e * Expressed as metaldebyde equivalentsEN-CAS Project # 90-0035 Lo Page 149 @ BV-CAS uathod Ho. m-2/04 © page 23 Soll control (e12") Attenuation = 2 EN-CAS Sazple ID #:_ B71890~c2 Metaidehyde Zquivalents Found: <0.02 ppm GC run # 37237, set uV2, dated 11/16/91 ® + Eepressed as’ netaldebyde equivalentsEN-CAS Project # 90-0025 Lo Page 150 e reid send 0, B27 me Ficuer 10 ‘Typical corematocran S041 Control ,+ 0.02 ppm Mataldehyde (6-12") Low Fortification Attenuation = 2 mune ke wey a6. 981 earazeas, oas8 9.72q@— Inomer 1 acetaldehyde sere ve.ags ins Toomer 2 acataldekyis 2-H EN-CAS Sample DD #: Bs1890-s2 Metaldehyde Equivalents Recovered: 109% Gc run § 37237, set v2, dazed 11/16/91 @ * Expressed as netaldehyde equivalentsEN-CAS Project # 90-0035 Lo Page 151 e BOs method Ho. mIC-2/91 rage 33 FIGURE 12 ‘Typical chromatesras Soil Control + 1.0 ppm Metaldehyde (e-a2") High Fortification Attenuation = 27 meee 3 omy iss issn raiatees EN-CAS Sample ID f: 271990-s5 Metaldehyde Equivalents Recovered: 117% Go run #37237, set MV2, dated 11/16/91 e * Expressed as aetaldehyde equivalentsEE-CAS Project # 90-0035 LO Ba Page 152 e HCAS Method No. 21C-2/91 rere 36 ‘FIGURE 12 ‘Typical chromatoome Soil control (a2-28") Attenuation = 2 fue enw a2, agen enesze 9.300 3-970 omer 2 meataldabyia ZN-CAS Sample ID f: Zyi891-c1 Metaldenyde Bquivalents Pound: <2-02 ppa (0.0072 pm) Fun # 37239, set MV3, dated 11/17/91, e + Bepreased as uetaldehyde equivalentsEN-CAS Project # 90-0035 Lo Page 153 N-CAS Method Mo. ENC-2/91 Page 37 ‘Typical _chrosatecras Soil Control + 0.02 ppm Netaldehyde tows a2 ev 17s 9k eaeizese stat eee EN-CAS Sample ID #:_ E71891-52 Metaldenyde Equivalents Recovered: 91% Gc run # 37239, set mV3, dated 11/17/91 * Expressed as netaldebyde equivalentsEN-CAS Project # 90-0035 LO Page 154 YI-CAS Method Ho. mNC+2/91 Page 38 Ficus 16 qTynical chromatocres Soil Control + 1.0 ppm Metaldebyde (ua-28") High Fortification Attenuation © 2” fun ern wee are igen tasenez treet Fas tae 272d bemer 1 senate ZH-CAS Sample ID #:_ 21891-s6 Metaldebyde Equivalents Recovered: 94 Gc run #37239, set MV3, dated 11/17/92 + Expressed as netaldenyde equivalentsEN-CAS Project # 90-0035 Lo Page 155 + EMeCAS Method Ko. ENC-2/91 Page 39 vous 15 ‘Typical cc Calibration curve for 0.01 ~ 0.10 ng METALDEM.JE SOIL 12-18" PEAK HEIGHT (Thousands) oO. O 0.01 0.02 0.03 0.04 0.05 0,06 0.07 0.08 0.09 0.1 ‘NG INECTED Gc run #37239, sat MV3, dated 11/18/91EN-CAS Project # 90-0035 Lo Page 156 EN-CAS Method No. ENC-2/91 Page 40 ‘FIGURE 16 ‘Typical ¢c Calibration Curve for 0.10 - 5 ng METALDEHYDE SOIL 12-18" PEAK HEIGHT (Thousands) 300+ 200+ 400 ou Gc run # 37239, set MV3, dated 11/18/91