1 15 1 https://www.nal.usda.gov/exhibits/speccoll/files/original/c0208aa220ef749ca2646df751f7681e.pdf c1fd91136b3c2e8c6913738e5cb32d39 PDF Text Text Item ID Number Author Brown, Marry D. Corporate Author Final Report/Article TltlB Report: Herbicide (2,4-D) Applicator Exposure Measurements Journal/Book Title 000 Year ° Month/Day Color n Number of Images 48 DBSCrlptOn Notes Supported by USDA Agreement No. USDA-TPSU-RU-0191 and New Jersey Agricultural Experiment Station Project No. NJ04502. Tuesday, May 15, 2001 Page 1466 of 1514 �FINAL REPORT HERBICIDE (2,4-D) APPLICATOR EXPOSURE MEASUREMENTS Harry D. Brown 106 Administration Bldg. New Jersey Agricultural Experiment Station Cook College, Rutgers University New Brunswick, N.J. 08903 Supported by USDA Agreement No. USDA-TPSU-RU-0-191 and New Jersey Agricul tural Experiment Station Project No. NJ04502. �INTRODUCTION The phenoxy herbicides have been used to control broad-leafed weeds in crops, water sources, forests, pastures, range lands, gardens, lawns, urban and industrial sites. Because of the efficiency, economy and safety, the phenoxy herbicides, especially 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and 2,4-dichlorophenoxyacetic acid (2,4-D), besides other herbicides, remain essential for agricultural and other uses (Diaz-Colon and Bovey, 1976). Use of 2,4,5-T and 2,4-D on a large scale to defoliate vegetation has resulted in controversy about the fate of the chemicals. Many of the hazardous properties ascribed to 2,4,5-T are to a lesser extent shared by 2,4-D. As a result, a number of papers dealing with the fate in the environment, toxicological effects on living organisms (Diaz-Colon and Bovey, 1976; Sauerhoff et al., 1977; Piper et al. , 1973) have been published. Cooper (1974) compared 2,4,5,-T and 2,4-D. He has reported that commercial samples of 2,4-D in mice were teratogenic and embryotoxic. Increasing controversy about their safe use has raised doubt about their utility in achieving vegetation management objectives. A Dow Chemical report on Forestry Applicators exposure to 2,4-D has recently been published (Levy, 1980). Nonetheless, in total, only meager information regarding human (e.g. gardeners, small farmers, etc.) exposure to 2,4-D is available. This study reports an investigation conducted under conditions characteristic of actual use pattern to determine the quantity of body surface contact of applicator with the commercial 2,4-D formulation and the quantity of the chemical measurable in serum and urine. MATERIALS AND METHODS Applicator Eleven healthy young male (ages 19 to 31 years) human volunteers were selected and were examined by the University physician prior to the work. Each subject was examined for blood pressure, pulse rate and body temperature on the day of work, both before and after the work period, and similar examinations were made every morning for the next three days (Table 1). Sterile gauze pads (size 0.67 sq. ft.) were attached on the back and chest of each volunteer. A paper cap (area 0.44 sq. ft.) was used to cover the head. Location The study was carried out either on the campus of Rutgers University or near the campus in New Brunswick, New Jersey during the period from May, 1981 to August, 1981. The location was covered with weeds and grass. Each area was roughly between one-half to one acre. Spray Material and Equipment The spraying material consisted of DMA-4 concentrate (obtained as a gift from Dow Chemical, U.S.A.) which contained 3.8 Ib acid equivalent/gal 2,4-D. The herbicide was mixed with tap water prior to use and the resulting mixture was applied at a rate of approximately 1 gal/30 min. by a hand sprayer held at hip level. A DuPont air sampler was attached to the applicator and was kept running during the spraying operation. The areas were covered by spraying six gallons in about three hours as per label instruction. �Weather Conditions Weather conditions during this study were moderate, temperature ranging between 65° to 78° F with humidity from 50 to 89%. Wind was fairly calm throughout this study except on one occasion, when it gusted to 10 miles per hour. Sample Collection for Analysis • To ascertain body surface exposure, gauze pads and caps were collected immediately after the work and washed in methanol. Air filter absorbents were also washed in methanol. Residue separated by methanol was evaporated to dryness and then treated with 15% BF, in methanol. Finally, it was extracted with n-hexane (Yip, 1975). Blood samples were drawn before the work and at the end of the day's work. Three more blood samples were drawn on the following three mornings. Overnight pooled urine samples were collected before the work and the day's collections were made during and/or after the treatment. Then the pooled urine samples were collected twice a day for the next 4 to 7 days. Blood sera and urine samples were treated, extracted and analyzed by the method of Sauerhoff et al.5^ (1977). Vegetation samples and soil samples from the treated area were collected before the treatment and then once every 24 hours for the next 4 days after the treatment. Samples were treated and analyzed by the methods of Davidonis et al. (1980) and Olson et al. (1978). Analytical Procedures 2,4-D residue extracted in n-hexane was analyzed on a pas chromatograph (Tracer Model No. 560, equipped with electron capture detector and a six foot glass column packed with 3% OV-101 on 80/100 supelcoport). Clinical determinations on blood serum and urine were made through the automated procedures of a commercial diagnostic laboratory. RESULTS AND DISCUSSION The various data are summarized in tables 1-13 and figures 1-22. At the end of a day's work, an applicator had an average 2,4-D residue of 49.39 yg/sq. ft. on the back, 52.41 yg on the head. The average value of 2,4-D residue on vegetation in the exposed areas was 27.93 yg/g of tissue at the end of the working day (i.e. first day). The residue fell to 8.57 yg/g tissue after four days. On the other hand, the average soil residue value was 2.68 yg/g soil at the end of the first working day, which Increased to 5v12*yg/soil (87.6%) on the fourth day. The average base level for phenolic compounds in serum was 70.7 ng/ml at zero level exposure. Immediately after the day's work, the value went up to 165.3 ng/ml and again went up to 267.7 ng/ml on the second. The difference between the base level and the measured level we take to be a specific measurement of the test material, 2,4-D. The value came down to 123.7 ng/ml on the third day. The average amount of phenolics ranged from 3.45 yg (zero level exposure) to 7.85 yg (fifth day) for the 12 hours of pooled urine. *Appendix A is a more extensive discussion of methodology. �Some changes were noticed in clinical analysis made on blood aerum and urine. Analysis of 2,4-D retained by the air monitor filter revealed detectable residue. The amount of air drawn through the filter was calculated to be 56.6 to 84.96 liters, and the calculated 2,4~D residue was 43.1 to 60.1 parts per trillion. Results of this study indicate that there was a considerable amount of 2,4-D in the air, on vegetation and soil surface of the treated areas. An appreciable quantity of 2,4-D also landed on the body surface, All serum and urine samples showed measurable 2,4-D residue. Statistical Analysis * The clinical residue and weather data were critically scrutinized by both linear models and regression models. The primary emphasis of the analysis was to identify the parameters that recorded any significant change. This was accomplished by subtracting the baseling value from the observed value and then dividing it by the baseline. Analysis of the resulting values revealed highly significant (a=0.01) changes in the serum creatinine and total protein levels, and significant ( H . 5 change in the urine specific gravity. It was also seen o)0) that the total protein and BUN/creatinine were positively correlated with the residue on the chest of the volunteers and with their height. When the peak residue in the body fluids was related with the external factors, it was seen that the serum residue was positively correlated with the 2,4-D concentration in the air and the amount of chemical that had landed on the head and back. Likewise, the urine residue was positively correlated with the air volume and the height of the volunteers. Precipitation (rainfall-high humidity) was negatively correlated with the serum residue, presumably due to decreased chances of vaporization and subsequent inhalation. *Analysis done by Dr. Robert Trout, Statistician for the NJAES. CONCLUSION Our observations, therefore, indicate that during normal working conditions, an appreciable amount of 2,4-D can enter into the applicator's body through inhalation and/or through body surface (dermal) absorption. Lengthy retention times have been observed in some subjects (Table 6). These findings are consonant with the report of Levy et al. (1980) for foresters. Clinical physiology data obtained here are consistent with long retention times for this chemical. We surmise, but have no specific evidence, that the 2,4-D molecule or some portion (phenolic), binds to serum protein and is eliminated as a function of the turnover of the protein molecules. Other than the clinical (chemical) changes noted above, we have seen no pronounced adverse effect of 2,4-D exposure. REFERENCES 1. Sauerhoff, M.W., Braun, W.H., Blau, G.E. and Gehring, P.J. (1977). The Fate of 2,4-Dichlorophenoxyacetic Acid (2,4-D) Following Oral Administration to Man. Toxicology 8:3-11. 2. Piper, W.N., Rose, J.Q., Lang, M.L. and Gehring, P.J. (1973). The Fate of 2,4,5-Trichlorophenoxyacetic Acid (2,4,5-T) Following Oral Administration to Rats and Dogs. Toxicol. Appl. Pharmacol. 26:339-351. �3. Levy, T.L. (1980). Determination of 2,4-D Exposure Received by Forestry Applicators. Dow Chemical, U.S.A. Report. October, 1980. 4. Diaz-Colon, J.D. and Bovey, R.W. (1976). Selected bibliography of the phenoxy herbicides. I. Fate in the Environment. MP 1303. Texas Agric. Exp. Stn., College Station. 5. Cooper, P. (1974). Food Cosmet. Toxicol. 12:418-421. 6. Davidonis, G.H., Hamilton, R.H. and Mumma, R.O. ( 9 0 . Comparative Meta18) bolism of 2,4-Dichlorophenoxyacetic Acid in Cotyledon and Leaf Callus from Two Varieties of Soybean. Plant Physiol. 65:94-97. 7. Olson, B.A., Sneath, T.C. and Jain, N.C. ( 9 8 . Rapid Simple Procedure 17) for the Simultaneous Gas Chromatographic Analysis of Four Chlorophenoxy Herbicides in Water and Soil Samples. J. Agric. Fd. Chem. 26:640. 8. Yip, G. (1975). Analysis for Herbicides and Metabolites. J. Chromatographic Sciences 13:225-230. �CLINICAL XXX DATA �Table 1 PHYSICAL EXAMINATION Temp ( F °) Blood Pressure Pulse Rate Applicators 0 X 1 2 3 0 X 1 2 3 0 X 1 2 3 No. 1 98.0 98.4 97.8 98.4 98.4 128/74 114/70 120/78 120/60 120/78 76 60 80 78 72 No. 2 97.6 98.0 98.0 98.0 97.8 110/78 118/78 110/78 110/72 120/80 60 72 80 68 64 No. 3 97.4 97.6 97.8 97.0 97.8 112/82 120/70 110/74 118/6?. 120/76 60 72 60 61 64 No. 4 98.2 99.4 97.4 98.0 98.0 90/60 100/68 100/64 96/68 92/60 80 100 76 76 84 No. 5 97.6 98.8 98.0 98.2 98.2 112/74 124/80 120/82 120/80 110/80 72 84 84 80 84 No. 6 97.0 98.6 97.2 97.0 97.4 120/80 110/74 108/78 112/78 112/74 84 88 96 92 100 No. 7 97.4 98.4 97.0 97.0 98.2 104/64 100/60 110/64 120/64 110/70 78 78 80 78 80 No. 8 98.4 98.4 98.6 97.8 98.0 140/76 120/80 112/70 130/68 130/76 80 64 96 80 68 No. 9 98.6 98.4 97.0 98.0 97.2 110/70 110/70 110/80 120/80 120/74 60 60 68 80 64 No. 10 97.8 98.0 97.4 98.2 97.8 104/60 100/70 108/60 112/70 102/60 84 68 84 88 88 No. 11 97.4 9 . 97.2 97.2 97.0 86 120/68 110/70 110/80 112/70 110/76 80 80 80 68 80 Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment �Table 2 Detection of 2,4-D Residue from Gauge Patch (= Body Surface) yg/sq. ft. 1 Applicator No. Head Chest Back 1 8.55 7.81 5.10 2 14.06 11.68 10.60 3 7.06 3.37 3.77 4 222.20 171.50 99.63 5 34.22 94.53 41.16 6 141.80 111.50 74.23 7 76.16 68.39 46.64 8 16.99 13.56 11.86 9 19.12 14.58 10.29 10 18.55 31.46 4.46 11 17.80 14.95 18.63 �Table 3 2,*i-D Residue on Vegetation Treated Area 2,4-D yg/g Vegetation 0 X X+1 X+2 X+3 1 0.59 28.85 16.15 16.67 20.16 2 14.74 29.60 22. A3 4.32 2.13 3 00.00 25.35 7.70 1.92 3.44 2.47 X+4 Amount of 2,^-D on Soil Surface Treated Area 2,4-D -fjg/g Soil X+1 X+2 X+3 «* 3.70 1.90 2.80 7.30 - 3.58 4.21 0.72 1.59 7.25 - 0.00 0.13 0.15 0.27 0.80 0.93 0 1 1.24 2 3 X X X X X 0 X 1 2 3 1» = = = = = *s Pretreatment Posttreatment 1 day after treatment 2 days after treatment 3 days after treatment ^ days after treatment �Table 4 Amount of 2,4-D Residue and Endogenous Phenolic Compounds in Serum Applicator No. 1 2 3 k 5 6 7 8 9 10 11 0 2,4-D Residue (ng/ml ) X+1 X 75.1 826.6 114.4 170.1 131.5 112.8 112.3 54.0 93.5 157.8 56.3 221.9 208.3 123.5 44.5 3.5 29.0 19.0 224.4 00.0 00.0 443.2 538.3 19.1 50.2 7.8 496.9 862.9 119.3 55.7 41.8 93.3 55.6 X+2 311.0 250.1 74.6 92.4 169.9 94.3 194.7 43.6 43.0 58.8 27.9 X+3 . 112.3 102.3 479.6 84.2 245.1 674.6 178.6 83.4 25.7 36.3 114.8 • 0 = Pretreatment X - Posttreatment X+1 «= 1 day after treatment X+2 = 2 days after treatment X+3 = 3 days after treatment Note: Because phenoxy and other phenolic compounds have liquid chromatographic migrations similar to that of 2,4-D, there is a variation in the base for specific measurement of the test compound. �\0 Table 5 Amount of 2,4-D in Serum Applicator No. X i X+l 2,4-D (ng/rol) X+2 714.3 198.7 0 196.1 48.3 2 60.4 116.1 3 128.4 38.0 4 285.4 5 X+3 482.0 — 440.6 654.6 6 — — 113.6 386.1 — 188.8 466.3 7 179.9 74.8 — 150.2 8 15.6 52.2 40.1 9 21.2 12.8 14.0 10 74.3 39.8 — 17.3 11 55.6 27.9 114.8 Legend X = X+l X+2 X+3 Posttreatment = 1 day after treatment = 2 days after treatment = 3 days after treatment 134.1 79.9 �Table 6 Total Amount of 2,4-D and Endogenous ^henolic Comnounds Excreted in the Urine yg 2,4-D Excreted per 12 hours Applicator No. 0 1 12 2k 36 48 60 1 3.73 7.12 2.38 2.58 6.49 5.70 2.93 21.20 28.96 - - - - 2 3.22 7.17 1.22 2.85 6.00 5.75 4.46 6.44 - - - 3 2.85 4.25 10.00 8.87 1.75 0.60 1.33 10.79 1.09 4.07 - - - - 4 8.11 15.44 21.38 7.66 9.56 5 0.32 1.31 1.79 1.60 1.31 6 9.95 7 9.91 8 0.00 0.11 9 10 72 84 96 108 120 132 144 156 - - - - - - 30.55 24.87 13.09 6.23 - - - - 2.62 3.32 2.27 7.39 - - - - - I*. 56 15.62 5.04 23.39 13.87 6.28 8.46 13.16 10.81 15.25 2.73 3.12 2.89 0.91 5.00 8.84 - - - - - 0.57 2.77 1.34 3.46 2.84 3.08 5.30 1.97 1.49 2.68 0.00 0.00 0.00 0.00 0.75 0,00 0.85 2.46 8.28 6.57 1.46 4.09 1.42 2.10 3.73 2.21 4,31 4.47 0.00 1.9* 1.73 1.63 2.83 2.21 2.73 8.48 4.35 - 0 • Pretreatment 1 • Posttreatment - - - - - - - - - �Table 7 HEMATOLOGY WBC x 103 liters 0 X 1 RBC x 106 RGB (g/dl) 2 3 0 X 1 2 3 HCT (%) 0 X 1 2 3 0 X I 2 3 3. 1 6.0 7.1 6.3 7.0 5.5 4.78 j, 4.68 j, 4761 j, 4*50 j. 4.43 15.5 15.1 14.8 14.6 14.6 44.4 43.7 44.8 441.6 441.5 3. 2 5.7 6.8 5.9 5.2 5.2 4.99 5.06 5.15 4.85 4.89 15.9 15.6 16.2 15.0 15.3 43.2 43.8 44.7 42.0 42.4 3. 3 6.7 6.1 5.2 5.6 5.0 4*69 4*64 4*63 4.71 4*65 14.9 14.8 14.5 14.6 14.9 41*2 41*2 41*2 40*9 40*6 ! 3. 4 6.5 7.6 6.6 6.7 6.5 5.72 5.76 5.75 5.85 5.50 15.5 15.6 15.3 16.8 14.8 43.9 44.0 43.5 44.6 41*8 ! 3. 5 6.3 6.5 5.9 7.0 6.2 5.34 5.22 5.23 5.56 5.27 15.9 15.7 15.7 16.8 15.6 44.6 44.1 44.7 46.5 43.6 D. 6 6.7 7.8 6.7 8.4 6.9 5.39 5.23 5.46 5.43 5.03 16.8 16.9 17.1 17.8 16.2 47.6 46.3 48.5 47.8 44.9 3. 7 6.6 8.1 9.8 8.2 7.9 5.45 5.20 5.42 5.33 5.29 16.5 15.9 16.3 17.0 16.3 48.0 47.3 46.9 46.0 o. 8 8.3 8.3 6.2 6.3 6.1 4.97 4.75 4.99 4.99 5.04 8.1 7.8 5.21 4.76 5.07 5.42 5.14 15.8 413.2 44.1 6.0 15.7 413.9 42.6 6.2 15.7 413.2 42.8 6.1 15.2 412.4 42.9 o. 9 15.8 413.3 44.9 441.0 37*4 40*3 38.1 o. 10 7.8 7.5 8.1 7.6 7.7 4.97 5.06 5.33 5.14 16.4 17.1 16.3 4.8 6.0 6.0 5.9 4.99 4.81 4.95 5.11 14.8 14.4 14.9 14.9 45.0 43.0 45.0 4 - 4 - 4 41.2 37.8 39.3 47.1 4 40.7 45.9 5.0 15.8 •l 13*5 16.5 0. 4.80 44.54 11 Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment J. 3877 J. 35*3 X 4, 4- 42.0 J �Table 7 (cont.) HEMATOLOGY MCV ( 3 y) MCH (yyg) MCHC (%) Platelet Estimate Applicators 0 X 1 2 3 No. 1 93 93 97 92 94 32J4 32^0 No. 2 87 87 87 87 87 3lt7 30.8 3lts No. 3 88 89 89 87 87 31J8 3ll8 3ll3 No. 4 7*4 7*4 7*3 7*4 7*6 27.0 26t9 26*6 No. 5 81 81 82 81 83 29.6 No. 6 85 85 86 85 89 3l7l 32.2 31.1 32.6 No. 7 85 83 84 85 87 30.2 30.4 40.0 3lt? No. 8 87 87 87 86 87 30.9 No. 9 7*5 7*5 7*5 7*5 7*5 24*8 No. 10 91 90 90 89 90 No. 11 83 84 83 83 83 A 0 A X = Posttreatment 1-3 » Days after treatment A. 2 3 A. 0 X 1 2 3 0 X 1 2 3 A. 34.7 34.3 32.9 34.9 35.1 N N N N N 31.0 31^2 36.6 35.5 36?1 N N N N N 30.8 32lO 36.0 35.9 35.1 35.4 36;6 N N N N N 28.5 27.6 35.2 35.2 34.9 37t4 35.8 N N N N N 30.0 29.7 30.1 30.3 35.4 35.5 35.0 35.8 36.2 N N N N N 35.2 36J 3 Dec N N N N 31^4 34.2 35.2 34.3 35.9 35.7 N N N N 3ltl 30.4 30.6 30.1 36.3 36t4 Dec N N N N 25*3 25t2 33.9 34.6 34.6 33.9 34.0 N N N N N 3itg sitg site 3it2 so. 8 35.7 36.1 35.8 35.6 34.9 N N N N N 28.8 35.2 35.1 36.0 3etl 35.0 N N N N N A 0 = Pretreatment A, 1 32J9 A Legend X A 32?0 32J3 A A 35.6 36.0 A A 24*9 A A A 33.0 24*9 28.9 29.1 29.2 28.3 •f A 34.9 36l9 A 36^4 35.9 3ets 35.2 N �Table 8 BLOOD CHEMISTRY Glucose (mg/dl) BUN (mg/dl) Creatinine (mg/dl) Uric Acid (mg/dl) Applicators 0 X 1 2 3 0 X 1 2 3 No. 1 93 88 79 81 76 12 12 13 11 No. 2 78 83 63* 80 75 18 17 17 19 No. 3 93 94 92 95 87 21 21 22 No. 4 87 112+ 86 92 86 12 14 No. 5 197+ 149+ 166+ 163+ 149+ 13 No. 6 111+ No. 7 81 0 X 1 2 3 0 X 1 2 3 11 0.9 1.0 0.9 0.8 0.8 6.4 6.5 6.1 4.9 5.2 16 1.2 1.3 1.2 1.2 1.0 6.1 6.3 6.6 6.3 5.5 27+ 26+ 1.0 1.0 1.0 1.1 0.9 3.8 4.5 3.9 4.8 4.1 12 13 15 1.1 1.2 1.2 1.1 1.1 7.6 7.9 7.5 8.4+ 8.1+ 14 12 13 15 1.0 1.0 1.0 0.9 0.9 6.6 7.7 6.6 7.0 6.9 95 91 92 18 19 18 16 18 1.2 1.3 1.2 1.1 1.2 6.3 6.6 6.4 5.8 6.8 137+ 76 88 91 14 15 16 18 15 1.2 1.5+ 1.2 1.1 1.2 7.0 7.1 7.1 7.2 7.3 107 15 15 17 13 13 1.1 1.0 1.0 1.0 0.9 5.6 5.3 5.1 5.4 4.4 81 No. 8 122+ 102 127+ 81 No. 9 118+ 90 107 79 122+ 17 16 15 17 17 1.1 1.0 1.2 1.0 0.8 5.7 5.8 5.4 6.0 4.9 No. 10 101 84 134+ 99 140+ 15 15 16 15 14 1.0 1.1 1.2 1.1 1.0 6.3 7.0 7.3 6.9 5.9 No. 11 148+ 124+ 112+ 98 111+ 20 19 17 14 12 1.5+ 1.0 1.0 1.0 0.9 7.3 5.1 4.8 5.0 4.7 Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment �Table 8 (cont.) BLOOD CHEMISTRY A/G Ratio Globulin (g/dl) Calcium4"1" (mg/dl) Bun/Great Applicators 1 0 1 0 0 No. 1 2.7 2.6 2.6 2.3 2.3 1.7 1.8 1.8 1.5 2.0 13.3 12.0 14.6 14.5 14.2 4.3 4.3 4.3 4.5 4.2 No. 2 2.5 2.5 2.4 2.5 2.2 1.9 1.9 1.9 1.8 2.1 15.0 13.1 14.2 15.6 16.2 4.3 4.2 4.1 4.1 4.2 No. 3 2.4 2.4 2.3 2.4 2.2 2.1 2.0 2.2 2.0 2.2 21.0 21.0 22.0 24.5 28.5 4.4 4.5 4.5 4.3 4.5 No. 4 2.5 2.5 2.6 2.3 2.2 1.6 1.7 1.6 1.8 1.9 10.5 11.2 9.7 12.0 14.3 4.4 4.4 4.3 4.4 4.3 No. 5 3.2 3.5 3.4 3.2 2.8 1.4 1.3 1.3 1.4 1.7 12.4 13.5 12.8 13.6 16.2 3.8 3.8 3.9 4.0 4.0 No. 6 3.0 2.8 2.9 2.6 2.2 1.6 1.7 1.6 1.8 2.1 14.7 14.1 14.2 14.2 14.5 4.2 4.5 4.3 4.4 4.4 No. 7 3.1 3.1 2.9 2.7 2.5 1.6 1.4 1.5 1.6 1.8 11.7 10.1 13.6 15.7 12.7 4.2 4.2 4.3 4.4 4.3 No. 8 2.5 2.6 2.4 2.4 2.6 1.9 1.4 1.9 2.0 1.7 14.7 15.2 16.9 13.9 15.4 4.3 4.3 4.2 4.4 4.3 No. 9 2.8 2.7 2.5 2.7 2.5 1.6 1.7 1.8 1.7 1.7 15.9 15.6 12.6 17.1 "20.4 4.2 4.1 4.3 4.3 4.2 No. 10 2.8 2.7 2.8 2.5 2.6 1.6 1.8 1.8 2.0 1.8 16.0 13.3 13.4 13.3 14.1 4.2 4.2 4.2 4.4 4.2 No. 11 2.5 2.7 2.8 2.7 3.0 1.9 1.7 1.6 1.7 1.5 13.3 18.3 17.3 14.2 13.7 4.3 4.2 4.3 4.3 Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment 4.1 �Table 8 (cont.) BLOOD CHEMISTRY Applicators LDH (IU/1) SCOT (IU/1) ' Alk. Phos. ( U 1 I/) GGT (IU/1) T. Bill, (mg/dl) 0 X 1 2 3 0 X 1 2 3 21 160 175 201 140 164 23 23 22 20 22 23 209 224 2$7 2$1 235 16 16 15 25 24 26 151 161 157 177 172 10 10 11 16 16 15 136 152 157 144 164 24 20 23 20 29 28 154 167 153 150 148 70 26 23 23 36 23 166 153 176 146 114 117 28 33 23 26 24 160 148 152 4*4 46 45 31 32 31 20 20 152 154 6*4 80 67 68 32 40 34 33 40 183 96 111 103 99 41 32 34 23 26 58 58 31 32 32 22 24 0 X 1 2 3 0 X 1 2 3 No. 1 42 40 43 45 39 22 22 20 22 No. 2 79 79 80 82 79 25 26 24 No. 3 69 61 71 79 79 22 24 No. 4 64 62 63 63 63 11 No. 5 90 81 88 87 87 No. 6 75 68 76 75 No. 7 116 111 115 No. 8 4*6 5*1 No. 9 5*2 No. 10 6*9 No. 11 116 5*8 6*1 Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment ! 0 X 1 2 3 20 0.6 0.7 0.4 0.3 0.8 15 15 0.7 0.7 . . 0.7 08 0.6 11 12 12 0.4 0.6 0.5 0.7 0.4 24 24 23 21 0.5 0.5 0.7 0.6 0.4 36 36 36 39 38 0.2 0.2 0.5 0.5 0.5 145 16 15 16 14 14 0.9 0.8 1.0 its itl 136 129 15 17 14 14 14 0.6 0.9 0.3 0.2 0.2 157 151 154 13 14 13 17 15 0.9 0.7 0.9 0.8 0.5 201 191 2$8 197 10 14 21 18 17 itl 0.8 0.9 0.7 0.9 195 179 187 169 165 16 10 11 14 12 itl 0.8 0.9 0.9 0.8 213 189 202 191 159 10 10 10 13 13 0.5 0.6 0.6 itl 1?3 �Table 8 (cont.) BLOOD CHEMISTRY Cholesterol (mg/dl) T. Protein (g/dl) Calcium (mg/dl) Albumin (g/dl) Applicators 0 X 1 2 3 0 X 1 2 3 0 X No. 1 179 173 177 1(1)8 169 9.9 9.8 9.7 9.1 9.4 7.3 7.2 No. 2 223 221 215 219 175 9.8 9.6 9.3 9.5 9.3 7.2 7.2 7.0 7.2 6.7 4.7 No. 3 its its 151 lt7 10.2 10.4 10.3 10.0 10.2 7.4 7.3 7.3 5.0 4.9 5.0 4.9 4.9 No:. 4 170 180 170 172 172 .9.6 9.7 9.4 9.7 9.2 6.7 6.8 6.7 6.6 6.4 4.1 No. 5 202 222 218 218 208 9.0 9.4 9.5 9.5 9.4 7.7 8^2 7.9 4.4 4.7 No. 6 210 208 209 208 192 10.0 9.7 7.7 7.6 7.5 7.3 6.9 4.7 4.8 4.6 4.7 4.7 No. 7 154 152 its it? 150 10.3 9.9 9.8 9.6 stl 7.4 7.2 7.0 6.8 5to 4.4 4.3 4.3 4.4 No. 8 165 169 170 180 172 9.7 9.9 9.7 10.1 9.7 7.1 7.3 7.2 7.1 4.6 No. 9 153 160 163 163 itg 9.7 9.5 9.6 10.0 9.4 7.3 7.3 7.0 7.5 6.4 4.5 4.6 4.4 4.8 4.4 No. 10 158 168 170 168 155 9.6 10.0 10.0 10.5 9.7 7.3 7.6 7.6 7.5 7.3 4.5 No. 11 161 lt9 160 160 157 9.9 9.9 7.3 7.1 7.3 7.3 7.4 4.8 4.5 4.5 4.6 4.4 its Legend 0 = Pretreatment X = Posttreatment 1-3 = Days after treatment 10.6 10.2 10.1 9.6 9.5 9.9 9.9 1 2 7.1 ste 7.1 3 6.7 7.4 7.1 7.8 7.6 0 X 1 2 4.6 4.6 4.6 3t3 3 4.5 4.7 4.6 4.7 4.6 4.3 4.1 4.3 4.2 4.5 4.6 4.7 4.8 4.7 4.8 4.5 4.9 4.9 4.9 4.7 �Table 9 ACID PHOSPHATASE AND HAPTOGLOBIN HAPTOGLOBIN (mg/dl) ACID PHOSPHATASE (IU/1) Applicators 0 X 1 2 3 1 2 3 4 5 No. 1 0.2 0.3 0.0 0.0 0.0 22 17 20 21 25 No. 2 0.3 0.3 0.4 0.0 0.0 24 26 24 25 18 No. 3 0.2 0.1 0.1 0.0 0.0 71 89 87 82 85 No. 4 0.0 0.0 0.0 0.0 0.04 92 113 111 76 108 No. 5 0.0 0.0 0.0 0.0 Missing 109 105 108 99 122 No. 6 0.0 0.0 0.0 0.02 0.12 189 197 204 174 238 No. 7 0.0 0.0 0.0 0.0 0.16 87 95 85 118 134 No. 8 0.2 0.0 0.0 Missing Missing 79 157 81 91 72 No. 9 0.0 0.0 0.0 Missing 0.2 90 90 91 165 80 No. 10 0.0 0.0 0.0 Missing 0.1 95 154 173 86 161 No. 11 0.0 0.0 0.0 Missing 0.6 159 114 118 118 128 Legend 0 «= Pretreatment X = Posttreatment 1-3 «= Days after treatment �\<\ URINALYSIS Table 10 pH Applicators 0 X la Ib 2a 2b 3a 3b No. 1 5.0 6.0 5.0 8.0 6.0 8.0 5.0 5.0 6.0 No. 2 6.0 6.0 5.0 6.0 6.0 6.0 6.0 6.0 5.0 No. 3 5.0 8.0 6.0 6.0 8.0 5.0 7.0 5.0 6.0 No. 4 5.0 5.0 5.0 5.0 6.0 5.0 7.0 5.0 6.0 No. 5 7.0 6.0 6.0 6.0 7.0 6.0 6.0 6.0 5.0 No. 6 5.0 6.0 6.0 6.0 6.0 8.0 6.0 6.0 6.0 No. 7 7.0 6.0 6.0 5.0 7.0 5.0 5.0 6.0 7.0 No. 8 6.0 6.0 6.0 5.0 5.0 6.0 6.0 7.0 5.0 7.0 5.0 8.0 No. 9 6.0 6.0 6.0 5.0 5.0 5.0 6.0 6.0 6.0 6.0 6.0 6.0 No. 10 6.0 6.0 6.0 5.0 6.0 5.0 5.0 5.0 6.0 No. 11 6.0 6.0 6.0 6.0 5.0 6.0 6.0 6.0 5.0 Legend 0 = Pretreatment X « Posttreatment 1-7 = Days after treatment a « 7:00 a.m. collection b « 7:00 p.m. collection 4a 4b 5a 5b 6a 6b 7a 5.0 6.0 6.0 6.0 5.0 6.0 — �Table 11 \ppll:ators So. 1 Sto. 2 So. 3 No. 4 No. 5 No. 6 No. 7 No. 8 No. 9 No. 10 No. 11 0 Amber Clear Amber Clear Amber Clear Straw Cloudy Straw Clear Straw Cloudy Straw Clear Yellow SI. cloudy Yellow Clear Yellow Clear Yellow Cloudy URINALYSIS COLOR X la Ib Yellow Clear Amber Turbid Yellow Turbid Straw Clear Straw Cloudy Straw Cloudy Straw Clear Yellow Clear Amber Clear Yellow Clear Yellow Clear Yellow Cloudy Amber Turbid Amber Clear Straw Clear Amber Clear Yellow Clear Yellow Cloudy Yellow Clear Straw Cloudy Straw Cloudy Straw Cloudy Straw Clear Straw Hazy Straw Hazy Straw Clear Straw Clear Straw Clear Yellow Hazy Yellow Clear Yellow Clear Yellow Cloudy Yellow Clear Legend 0 = Pretreatment X = Posttreatment 2a 2b Yellow Yellow Clear Clear Yellow Yellow Cloudy Cloudy Yellow Yellow Clear Clear Straw Yellow Clear Cloudy Yellow Yellow Cloudy Hazy Straw Yellow Cloudy Hazy Straw Yellow Clear Clear Straw Yellow Cloudy Hazy Straw Yellow Cloudy Hazy Straw Straw Clear Clear Straw Yellow Clear Clear 1-7 = Days after treatment a = 7:00 a.m. collection 3a 3b Yellow Clear Yellow Clear Yellow Clear Yellow Clear Yellow Clear Yellow Cloudy Yellow Clear Yellow Clear Yellow Clear Yellow Clear Yellow Clear Yellow Clear Yellow Turbid Yellow Clear Yellow Cloudy Yellow Cloudy Yellow Cloudy Yellow Cloudy Yellow Clear Yellow Cloudy Yellow Clear Yellow Clear 4a 4b 5a 5b 6a . 6b 7a Yellow i Clear Yellow ;— Turbid Yellow —_ Turbid Yellow —;_• Cloudy Yellow Cl Dudy Yellow Cloudy Yellow Cloudy Yellow Yellow Yellow Yellow Yellow Yellow Yello Clear Hazy Cloudy Hazy Clear Clear Hazy Yellow Yellow Yellow Yellow Yellow Yellow Yello Clear Clear Clear Clear Clear Clear Hazy Yellow Clear Yellow Turbid 7:00 p.m. collection �Table 12 URINALYSIS SPECIFIC GRAVITY Applicators 4a 4b 5a 5b X la Ib 2a 2b No. 1 1.022 1.010 1.026 1.007 1.015 1.010 1.023 1.010 1.015 1.028 1.027 1.028 1.028 1.024 1.024 1.026 1.033 1.028 1.033 1.023 1.030 1.026 1.033 1.020 1.024 1.015 1.022 1.008 1.020 1.020 1.020 1.018 1.024 1.019 1.022 1.018 1.020 1.022 1.022 1. 016 No. 6 1.029 1.028 1.025 1.024 1.026 1.017 1.029 1.024 1.012 1.014 1.016 1.016 1.017 1.014 1.020 1.014 1.026 1.026 1.020 1.022 1.026 1.027 1.027 1.012 1. 015 1.017 1.023 1.024 No. 9 1.013 1.027 1.025 1.025 1.031 1.031 1.030 1.032 1. 015 1.030 1.025 1.020 No. 10 1.017 1.018 1.026 1.007 1.008 1.006 1.021 1.011 1. 018 No. 11 1.025 1.016 1.017 1.020 1.022 1.012 1.016 1.012 1. 025 1.026 1.015 1.023 1.023 1. 012 No. 8 1.026 1. 028 'No. 7 1.030 1. 016 No. 5 _— 1. 030 No. 4 7a 1. 029 No. 3 6a 1. 006 No. 2 6b — _. 0 Legend 0 = Pretreatment X = Posttreatment 1-7 = Days after treatment a = 7:00 a.m. collection b = 7:00 p.m. collection 3a 3b . — �Table 13 U « I * AL » I I S Appllcatort 0 1 l a II BACTERIA ?ew CRYSTALS Calcium Oxalete tPITH-. Occet. CRYSTALS vValet I t la CRYSTALS IPITHi Uric Rare acid. Urates Jb 3a CPITH: fPtTH' wnc-0-l 1-1 b-r Jo tfitiji. Occet. IPIIH: CRYSTALS Occas. k-» Cal- It BACTERIA BACTERIA CRYSTALS CPITNi CRYSTALS XBCiO-l WBC Few Few Calcium Few Urates IVTTHi Rare . Oxalate CRYSTALS TI ' CPITHj Urates CRYSTALS TT"^ 8-1 Cal- CRYSTALS clue 0-1 Calctum OxaUta 1 1 BACTERIA BACTERIA UBC:J-J Few Few rRYS~TAL$ CRYSTALS PROTEIN: clum Calcium ~\ Oulata Oxalata PROTEIN; (PITHs UBC:0-I Few RaTe Ful 1 0-1 VBC; 0-1 UBC:0-I £r*- WBC: 0-1 WBC:0-t tpTTHi V3C:0-I W)C:l-2 laVTTALS IFfTH: VBC:0-2 trTTrli WBC:0-1 BTTH: Few ~* Few ?S few CRYSTALS Celclio Few 11 IJ Oxalates. many amor- It EPITH, WSC:0-I few— CTiYTTALS CRYSTALS Hi>|c CRYSTALS CRYSTALS CRYSTALS Calcium acid. Calcium AmorFew Cal- OxaCalcium Ouclum phous late, Oxalate late. •rates Oxalate, Uretas CLUCOSE Few CLUCOSE •wny Trace amorTrace phous 0-1 WBC:0-I tf»C:0-l UBC:0-I UBC:0-t CRYSTALS EPITH: CTiV5TAts Calcium Few cat - few Oxalase, CRYSTALS Clum AmorCalcium Oxalate phous Oxa late- CASTS: 0-fc urates Few. byline Amorphous casts •rates tPITH; Occas. CRYSTALS CRYSTALS Phos• Few cel- phetes clum BLOOO: Oxalate. TI Few mucus threads WDC:0-I EPITH: Few UBCiO-t WBC:0-I UBC:0-I ViCiO-l UDC:0-t WBC:0-I tpTTH: IfTTH: IFTTB: Few Few FI BACTERIA Few CRYSTALS Urates tPTTn: msa in— w«C:0-t tTTTH: IF7TH: Rare CRYSTALS CRYSTALS Occas. liare Calcium Calcium Oxalate 2* Amorphous Urates. 2» Hucus VtC:l-l n w«c!o-i l^rt W!C:0-I t?ll«: Rare YSTALS re Calcium Oxalate, tPITHl . Occas. CRYSTALS WBC:0-I |ljre ' W8C:0-I IPlTH; Rare EPITH: CRYSTALS - . Occas. CRYSTALS Urates {PITH: WBC-.I-J CRYSTALS . Occas . CRYSTALS Urates acid. Urates Urates nvTTALs urn CRYSTALS WBC:I-J tfTTHs phOUS Occas. SediCRYSTALS ments Urates JLOOD-.I* *zr-— WBC:0-I CRYSTALS CRYSTALS EPITH: Calcium Occas. Full Oxalate BACTERIA Field itoJ: Amor•mount, phous SediFew ment mucus threads CRYSTALS - ' . . . . . . . . . . . . . . . . . . . . . . RBC:Rara ITtTH: Occas. Few awicus threads rcq< threads yjCup-l JSCj.O-1 tflTH: CPITHi Rare Rare BACTERIA. Full field MliOO tPITHL Rare W1C;0-I VlCiO-t ITTTH; Rare iTTfHi m phous Urates MICWft VtC:0-l VBCJM I?TlHj. PTfrl: lire Rare BACTERIA :TERIA . Im- S V»C:0-1 CRYSTAL!I f requenlt Calcium Oxalate,i I* IhlCUS 1* rtocuiI Threads Threads fit . BACTERIA WK:0-t IACTERIA PROTEIN BACTERIA V>C:0-I 1+ Hoderate P.BC:Rare Many TRYTTAIS Many HlYS~TALS CRYSTALS Calciun CRYSTALS BACTERIA Few • Celclum Calcium Oulate AmorCRYSTALS Oxalate. phous Oxalate Amor1* Mucus Phosthreads . phates phous Urates, Calcium Oxalate CRYSTALS YSTALS tPITH: CPITH^ tPITH; CRYSTALS tPITH: tPITH; OccaOcca- ' Moderate Calcium ! T P Calcium Iclim Few Oulate CRYSTALS slonal Oxalate, Oxalate BACTtRIA slonal Calcium CASTS; AmorOxalate Calcium phous Field CRYSTALS Oxalate Sediment Moderate PROTEIN; «t amount Calcium Oxalate. Few wfit1 f frequent S IPITH: CfltH: Rare Rare fRYSTALlHYSTAlS frequent Rare Calcium Calcium Oxalate Oxalate - t.| phous •rates amorphous 7a Occat. BACTERIA UBCil-J W»C:»-1 . ITTTH; Occas. CRYSTALS ilralts CRYSTALS Few BLOOD I* Urates Urates CRYSTALS PROTEIN Orates ITTTH: JP1 CRYSTALS r^ field amorphous •rates K »b «o» • VBC:0-I WBClO-l CTlTTALS (a amorewcus • threads. phous sperurates ewto- CRYSTALS CRYSTALS Urates >hos- Ik Sb WIT EPITH; CftWAlS • Occas. Lg. «Mt. CRYSTALS I CJTV amorphous few uric «rates, < few calcium mulata tPTTH; Occas. vn* mucus- . threads ten ia <*- VBC 'o-i +1 WBC: 1-2 Occas. BACTERIA IFfTHl <* VBC- clum Oulata i Ve VBCiO-l tMTH! Rare tPITH; tPITH; Rare Rare BACTtRIA Faw - tPITM: Rare UO&i. Bcce llonal tPITH: tPITH; Occa- CRYSTALS Calcium slonal Oxalate �Figure 1. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 1.0 _ 0.6 _ 0.4 0 X 1 2 3 ,- 0.8 Subject No. 1 _ <u CO C£ 0.2 . Day = Pretreatment • Posttreatment = 1 day after treatment = 2 days after treatment = 3 days after treatment �Figure 2. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 0 X 1 2 3 1.0 0.8 M <U Subject No. 2 0.6 cn Q 0.4 00 0.2 \_ 1 Day « » = = = Pretreatment Posttreatment 1 day after treatment 2 days after treatment 3 days after treatment �Figure 3. 1.0 2,4-D and Endogenous Phenolic Compounds Measured in Serum Subject No. 3 0 X 1 2 3 i- 0.8 0) CO 0.4 60 0.2 1 Day = Pretreatment = Posttreatment = 1 day after treatment = 2 days after treatment = 3 days after treatment �Figure 4. 2,4-D and Endogenous Phenoli: Compounds Measured In Serum 1.0 r. 0.8 0.6 CO a 0.4 00 0.2 I 1 Day Subject No. 4 0 = Pretreatment X « Posttreatment 1 = 1 day after treatment 2 = 2 days after treatment 3 = 3 days after treatment �Figure 5. 2,4-D rahd Endogenous Phenolic Compounds Measured in Serum 1.0 _ 0.8 0.6 0.4 0.2 1 Day Subject No. 5 0 = Pretreatment X = Posttreatment 1 = 1 day after treatment 2 = 2 days after treatment 3 = 3 days after treatment �Figure 6. 0.8 0.6 p 0.4 0.2 Subject No. 6 0 X 1 2 3 1.0 <u 2,4-D and Endogenous Phenolic Compounds Measured in Serum - = = = = = Pretreatment Posttreatment 1 day after treatment 2 days after treatment 3 days after treatment �Figure 7. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 0 = Pretreatment X = Posttreatment 1 « a day after treatment 2 * 2 days after treatment 3 = 3 days after treatment 1.0 0.8 to 0.6 en H 0 00 Subject No. 7 0.4 3. 0.2 1 Day �Figure 8. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 1.0 0.8 0.6 <u en 0.4 OC 0.2 1 Day Subject No. 8 0 » Pretreatment X = Posttreatment 1 = 1 day after treatment 2 = 2 days after treatment 3 = 3 days after treatment �Figure 9. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 0 « Pretreatment X = Posttreatment 1 = 1 day after treatment 2 = 2 days after treatment 3 = 3 days after treatment 1.0 0.8 g to Subject No. 9 0.6 0.2 1 Day �Figure 10. 1.0 0.8 S> 0.6 CO o sf «M tn 0.4 0.2 2,4-D and Endogenous Phenolic Compounds Measured in Serum Subject No. 10 0 X 1 2 3 » Pretreatment » Posttreatment = 1 day after treatment = 2 days after treatment = 3 days after treatment �Figure 11. 2,4-D and Endogenous Phenolic Compounds Measured in Serum 1.0 0.8 $ 0.6 V) C5 CM be 04 . 0.2 Subject No 0 X 1 2 3 11 Pretreattnent Posttreatment 1 day after treatment 2 days after treatment 3 days after treatment �Applicator No. 1 0 » Pretreatment X « Posttreatment Figure 12. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine 30 20 12 24 36 48 60 Hour 72 84 96 108 120 132 144 �30 Figure 13. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 2 0 «• Pretreatment X = Posttreatment 20 o •ato * 10 12 24 36 48 60 Hour 72 84 96 108 120 132 144 �30 Figure 14. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine 20 p L 10 Hour Applicator No. 3 0 - Pretreatment X = Posttreatment �Figure 15. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 4 0 = Pretreatment X = Posttreatment 30 20 . cd 4J o CM t>0 10 12 24 36 48 60 Hour 72 84 96 108 120 132 144 �30 Figure 16. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 5 0 - Pretreatment X = Posttreatment 20 JC 10 120 Hour 132 144 �Figure 17. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine 12 24 36 48 72 60 Hour 84 Applicator No. 6 0 Pretreatment X Posttreatment 96 108 120 132 144 �30 Figure 18. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 7 0 = Pretreatment X = Posttreatment 20 o H CM 00 X 12 24 36 48 60 Hour 72 84 96 108 120 132 i 144 �30 Figure 19. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine 20 rt jj o 6C 10 Hour Applicator No. 8 0 * Pretreatment X = Posttreatment �30r Figure 20. Amount of Total 2,4-B and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 9 0 * Pretreatment X » Posttreatment 20- «8 •U Q CM 00 10- X 12 24 36 48 60 72 Hour 84 96 108 120 132 144 �30 r Figure 21. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine Applicator No. 10 0 = Pretreatment X - Posttreatment 20 O H (50 3- 10 I 12 24 36 48 60 72 Hour 84 96 108 120 132 144 �30 r Applicator No. 11 0 » Pretreatment X = Posttreatment Figure 22. Amount of Total 2,4-D and Endogenous Phenolic Compounds Excreted in Urine 20 10 12 24 36 48 60 72 Hour 84 96 108 120 132 144 �MS* Appendix Methodology for 2,4-D Analysis �Methods for Analysis of 2,4-D In order to select the most recent analytic techniques for 2,4-D assay in urine, serum, gauze patches, vegetation and soil samples, a literature survey was done. The following methods were used in our investigation: a. Urine: Out of the total volume of urine samples, 10 ml was taken and 2 ml of IN HC1 was added to it. The acidified urine was then mixed with 5 ml of diethyl ether and was shaken for 10 minutes on a Buchler-Vortex -evaporator (Vortex setting was kept at 5). This extraction method was followed three times. The ether extracts were pooled together and evaporated to complete dryness at a temperature not higher than 45°C. The dry residue was dissolved in 1 ml of methanol first and then mixed thoroughly with 1 ml of 15% solution of BF3 in methanol. The resultant mixture was heated in a water bath at 70°C for 10 minutes for complete methylation. The methylated solution was cooled and again extracted thrice with 1 ml of n-hexane. N-hexane extracts were either injected (1 yl) immediately in a G.C. or stored at -20°C. b. Blood Serum: One ml of isolated serum was diluted with 4 ml of distilled water and then acidified with 1 ml of IN HC1. The acidified serum was then treated (i.e. extracted and methylated) in exactly the same manner as was the urine. One yl of n-hexane extract was injected in a G.C. c. Gauze Patches: Immediately after the day's work, the patches (from the front, back and head cover) were soaked separately in 100 ml of methanol and kept overnight at 4°C. After washing the patches thoroughly with methanol, 50 ml of the wash (from each group) was evaporated to dryness at less than 45°C temperature. The dry residue was dissolved in 1 ml of methanol and then methylated and finally extracted as above. The methylated compound, however, was attracted three times with 4 ml, 3 ml and finally 3 ml of n-hexane. The pooled extractant was diluted when needed before G.C. analysis or stored as before at -20°C. d. Air Filter: Each air filter was washed with 50 ml of methanol immediately after the day's work and was kept at 5°C overnight. Clear methanol was removed and the filter was further rinsed twice with 10 ml of methanol. The pooled methanol was treated in the same way as the gauze patches. e. Vegetation: Ten grams of freshly collected leaves were chopped and homogenized in 50 ml of 95% hot ethanol. The homogenate was centrifuged at 5,000 rpm in a fixed angle rotar for 30 minutes. Supernatant was carefully removed and saved. The pellet was thoroughly washed with 50 ml of 80% hot ethanol and recentrifuged three times as before. Each supernatant fraction was pooled together in a round bottom flask and was evaporated to bring the final volume to 20 ml. The concentrated supernatant was adjusted to pH 3 by adding exactly 10 drops of H3PO^. This acidified solution was extracted first with 50 ml of diethyl ether and then with 25 ml and finally with 25 ml of diethyl ether. The total extractant was evaporated to dryness under a current of nitrogen at 45°C. The dry residue was again extracted three times with 3ml, 1 ml and 1 ml of methanol. The combined methanol extract was treated with 1 ml of 15% BF2 in methanol and kept under a nitrogen atmosphere in a water bath at 70°C until concentrated to 2 ml. This methylated solution was further extracted with nhexane as described above. N-hexane was diluted when needed for G.C. analysis. �f. Soil: Fifty prams of soil sample was mixed with 40 ml of IN H^SO, to prepare a slurr. To the acidified soil, 45 ml of diethyl ether was added and was taken in a separating funnel for extraction. The separating funnel containing the soil sample was shaken for a total of fifteen minutes, but for not more than one minute at a time. The solvent layer was decanted and passed through glass wool and the aqueous layer containing soil particles was rejected. The clear solvent layer was then mixed with 25 ml of IN NaOH and shaken vigorously in a separating funnel for 1 minute. The organic layer was rejected and the alkaline aqueous layer was cleaned by partition method with 25.ml chloroform. The clean aqueous layer was further acidified with 1 ml of cone. l^SO^ and extracted with 25 ml of ether. The ether extract was dried completely and redissolved in 5 ml of methanol. Methylation and final n-hexane extraction were made as in the case of the vegetation samples. The following gas chromatographic conditions were used: Detector, Electron capture Ni 63 temperature: 350°C Gas: Methane: Argon (5:95) Flow rate through the column 25 ml/min Sample volume injected 1 yl Column material 3% 0V 101 on 80/100 supelcoport Length of the column 6 ft I.D. of the column 2 mm Column temperature 187°C Injection port temperature 200°C � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 055 Folder The folder containing the original item. 1466 Series The series number of the original item. Series III Subseries II Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Brown, Harry D. Title A name given to the resource Final Report: Herbicide (2,4-D) Applicator Exposure Measurements Subject The topic of the resource human testing agricultural exposure industrial exposure ao_seriesIII