CHLORPYRIFOS JMPR 1972 IDENTITY Chemical name O,O-Diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate Synonyms Dowco(R)179, ENT 27311, OMS 971, DURSBAN(R) Structural formulaMol. Wt. 350.6 Empirical Formula C9H11Cl3NO3PS Other relevant chemical and physical properties State: white, granular, crystalline solid Melting point: 42 - 43.5°C Vapour pressure: 1.87 × 10-5mm Hg (at 25°C) 8.87 × 10-5mm Hg (at 35°C) Solubility: readily soluble in acetone, benzene, chloroform, methanol and iso-octane; slightly soluble (0.4 ppm) in water. When chlorpyrifos was exposed to UV light or to sunlight, it underwent hydrolysis in the presence of water to liberate 3,5,6-trichloro-2-pyridinol, which underwent further decomposition to diols and triols and ultimately cleavage of the ring to fragmentary products (Smith, 1968). Hydrolysis in water occurs least readily at about pH 6 and very readily above pH 8. The technical product is supplied in a variety of concentrations and formulations (emulsifiable concentrates, dusts, wettable powders and granules). Purity: 99.5% EVALUATION FOR ACCEPTABLE DAILY INTAKE BIOCHEMICAL ASPECTS Absorption, distribution and excretion As with other phosphorothioate esters chlorpyrifos is rapidly absorbed, metabolized and excreted by mammals following oral administration. When 36Cl chlorpyrifos was fed by intubation to rats as a single sublethal dose (50 mg/kg), the radioactivity was eliminated rapidly (90.1% in 26 h) via the urine (90%) and faeces (10%). Products excreted were identified as 36Cl 3, 5,6-trichloro-2-pyridyl phosphate (75-80%), 3,5,6-trichloro-2-pyridinol (15-20%) and traces of chlorpyrifos. The pyridyl phosphate was indicated to be a transient intermediary in the hydrolysis of chlorpyrifos to the pyridinol moiety. Analyses showed no appreciable residue in any tissue except fat, which showed temporary retention of radioactivity (halflife of ca 62 h) identified principally as chlorpyrifos (Smith et al., 1967b). This study suggests that desethylation is a significant metabolic pathway in rats. An additional study in the rat involving administration of 14C chlorpyrifos by single dose intubation (19.1 mg/kg) essentially verified the earlier result (Branson and Litchfield, 1971a,b). In this study, as well as in an in vitro study involving breakdown of chlorpyrifos by rat liver microsomes (Branson and Wass, 1970), 3,5,6-trichloro-2-pyridinol was identified as the only major metabolite. A study in cows involving feeding of chlorpyrifos in the diet at 5 ppm for four days indicated that the major pathway for degradation was by hydrolysis to the pyridinol, as no desethyl phosphate esters were found. In addition no chlorpyrifos or metabolites were found in milk (Gutenmann et al., 1968). Studies of 3,5,6-trichloro-2-pyridinol, the major enolic metabolite of chlorpyrifos, have shown absorption, distribution and excretion in the rat following single oral administration to be similar to that reported for the parent compound. The metabolite is rapidly absorbed and excreted primarily in the urine and faeces. Small tissue residues (<0.3 ppm) were present and occurred mainly in the biological systems involved with urinary excretion, i.e. liver, kidney and blood. Unlike chlorpyrifos, residues of the pyridinol in fat were trivial, which is consistent with its much greater polarity (Branson and Litchfield, 1971a,b; Smith et al., 1970). Biotransformation The metabolic fate of chlorpyrifos has been investigated in plants (Smith et al., 1967a, 1967b) and data indicate an oxidative and/or hydrolytic breakdown yielding des-mono-ethylchloropyrifos; desethyl chlorpyrifos; 3,5,6-trichloro-2-pyridinol and further degradation products. This is discussed further in the section on "Fate of residues." Effects on enzymes and other biochemical parameters Chlorpyrifos is a cholinesterase inhibitor affecting this enzyme in brain or blood, although differing in its response with different animal species. In vitro studies have shown the oxygen analogue of chlorpyrifos to be 106 times more active as an inhibitor of cholinesterase than chlorpyrifos (I50 values: chlorpyrifos approx. 2.5 x 10-3M; oxygen analogue approx. 2.5 x 10-9M) (Smith, 1966). Equimolar concentrations of chlorpyrifos administered to dogs and monkeys affected RBC cholinesterase in monkeys while having almost no effect on RBC cholinesterase in dogs. Plasma cholinesterase is the most sensitive indicator of exposure to chlorpyrifos. In dogs and monkeys administered subacute levels of chlorpyrifos, plasma cholinesterase was maximally inhibited within 8 hours following exposure. Recovery in dogs was complete within 24 hours while with monkey, plasma cholinesterase in monkeys remained depressed at 48 hours (Coulston et al., 1971). TOXICOLOGICAL STUDIES Acute toxicity of the metabolite 3,5,6-trichloro-pyridinol has been studied in rats and mice. Results are summarized in Table 1. Signs of poisoning with this metabolite include facial paralysis with dyspnoea and slight hypersalivation and exophthalmia (mice only) (Gerbig and Emerson, 1970 a,b). Special short-term studies on metabolites Rat Groups of rats (10 males and 10 females/group) were fed the metabolite of chlorpyrifos, 3,5,6-trichloro-2-pyridinol, in the diet for 90 days at dosage levels of 0, 100, 300, 1 000, 3 000 and 10 000 ppm. Diuresis was observed at 3 000 ppm. At 10 000 ppm a reduction in body-weight was observed which reflected a general retardation in growth in both sexes. No effects were observed on mortality, terminal haematology or gross and histological examination of tissues (Beatty and McCollister, 1964). Dog Groups of dogs (3 males and 3 females/group, 4 of each sex were controls) were fed the metabolite of chlorpyrifos, 3,5,6-trichloro-2-pyridinol, in the diet for 91 days at dosage levels of 0, 1, 3, 10 and 30 mg/kg/day (Emerson and Gerbig, 1970). Another study utilized groups of dogs (2 males and 2 females at the higher level and 4 male and 4 female at the lower level) which were fed the metabolite at dietary levels of 26 and 80 mg/kg/day for 93 days (Copeland, 1964). At 80 mg/kg the metabolite produced abnormalities including growth depression in females, increased spleen/body-weight ratios and histological changes in the liver. At 26 mg/kg an elevated SAP accompanied by equivocal changes in the liver was observed. At 30 mg/kg increased SAP, SGOT and SGPT values were observed in the second study. The increased biochemical parameters were noted in 1 of 3 dogs examined at 10 mg/kg. Hepatic lesions noted in 1 dog at 10 mg/kg and 1 dog at 30 mg/kg did not appear to be chemically induced. No effects were noted at 3 mg/kg/day on growth, behaviour, haematology, blood biochemistry, urinalysis or gross and microscopic examination of tissues. TABLE 1 Acute toxicity of the metabolite 3,5,6-trichloropyridinol Species Sex Route LD50 (mg/kg) Reference Rat M Oral 794 Gerbig and Emerson, 1970a F 870 Mouse M Oral 380 Gerbig and Emerson, 1970b F 415 Duckling Groups of Peking ducklings (10 per group, 20 used as controls) were fed the metabolite at dosage levels of 0, 1, 3, 10, 30, 60, 100, 300 and 1 000 ppm for 3 weeks to determine cataractogenicity. Growth retardation was evident at 1 000 ppm, and 3 of 10 ducklings had elevated prothrombin values. Levels of 300 ppm and below produced no effect on these parameters and no cataractous changes were observed at any level (Molello and Sharp, 1968). Special studies in neurotoxicity Hens (3 DeKalb leghorns/group) were administered chlorpyrifos orally at dosage levels of 40, 75, 100 and 150 mg/kg and observed for 27 days. Mortality occurred at the higher levels. Surviving birds showed no signs of delayed ataxia or paralysis. Crufomate administered as a positive control (1 000 mg/kg) produced clinical signs of paralysis (Stevenson, 1966). Special studies on potentiation Rats administered subacute oral dosages (1/2 the LD50 value) of chlorpyrifos in combination with malathion or dichlorvos showed slight evidence for potentiation of the acute toxicity. A three-fold increase in the acute toxicity was noted with malathion, but this was judged to be of no practical significance (Dow Chemical Co., 1964; Norris, 1970). Special studies on reproduction and teratogenicity Groups of rats (10 males and 20 females/group with 20 males and 10 females/group as controls) were fed chlorpyrifos in the diet for one generation (two litters) at dosage levels of 0, 0.03, 0.1 and 0.3 mg/kg and for two generations (two litters per generation) at dosage levels of 0, 0.1, 0.3 and 1.0 mg/kg. The F3b litter was used in a teratology study where the parents were fed dietary levels of chlorpyrifos until day 6 - 15 of organogenesis when the dietary supplement was removed and chlorpyrifos administered by gavage once daily. After day 15 chlorpyrifos was again administered by dietary supplement until day 20 when the females were sacrificed, foetuses removed by caesarean section and viscera and skeletal structure examined. Neonatal mortality was somewhat higher in F2a, F2b and F3a litters at 1.0 mg/kg. No other significant effects were observed on fertility, reproduction or lactation indices or on teratological examination of F3b pups as a result of feeding chlorpyrifos at levels up to 1.0 mg/kg/day. Cholinesterase depression was observed at levels of 0.3 mg/kg and above, primarily in plasma (Thompson et al., 1971). Acute toxicity The acute toxicity of chlorpyrifos has been studied in several animal species, and the results are summarized in Table 2. TABLE 2 Acute toxicity of chlorpyrifos in different animal species Species Sex Route LD50 (mg/kg) Reference Rat M Oral 118 - 245 Taylor and Olson, 1963; Gaines, 1969; Tucker and Crabtree, 1970 F Oral 82 - 135 Taylor and Olson, 1963; Gaines, 1969 Mouse M Oral 102 Coulston et al., 1971 - i.p. 31 USAEHA, 1968 White footed - Oral 64 Kenaga, 1967 Mouse Rabbit - Oral 1 000 - 2 000 Taylor and Olson, 1963 Cavy M Oral 504 Ibid. Goat F Oral 500 - 1 000 Tucker and Crabtree, 1970 Chukar F Oral 61.1 Ibid. M Oral 60.7 Ibid. Bullfrog M Oral >400 Ibid. Chick M Oral 25 - 35 Taylor and Olson, 1963; Stevenson, 1966; Sherman et al., 1967; Brust Chicken - Oral 32 - 63 et al., 1971 The LD50 in a number of avian species has been determined by Tucker and Crabtree (1970). The toxicity values range from 8 mg/kg in male ring necked pheasant to 167 mg/kg in mallard ducklings. Signs of poisoning with chlorpyrifos are typical of cholinesterase inhibiting compounds. Mortality following acute oral dosing may be delayed for as long as nine days in some species of animals (Tucker and Crabtree, 1970). Chlorpyrifos administered to the conjunctival sac of rabbits produced transient redness of the conjunctiva. No corneal injury was observed (Taylor and Olson, 1963). When applied to intact or abraded abdominal skin of rabbits daily for up to 2 weeks no significant dermal damage was observed. Prolonged contact produced burning and hardening of the skin, slight swelling and mild hyperemia which returned to normal within three weeks. Chlorpyrifos administered dermally as a technical product to rabbits in an undiluted form produced no untoward physiological effects. When administered as a 50% solution in dipropylene glycol methyl ether, the same dose (2 000 mg/kg) was rapidly absorbed and resulted in death in four out of six animals when applied to either intact or abraded skin (Taylor and Olson, 1963). Chlorpyrifos did not induce sensitization when examined on the guinea pig (USAEHA, 1968). Dogs and sheep were exposed for a 4-hour period to a thermal fog or liquid aerosol at a concentration of 4 or 8 mg/cu ft. Mild plasma cholinesterase depression in dogs at the higher dose was the only effect noted (Dow Chemical Co., 1966a). Five female rats were exposed for 7 hours/day to an atmosphere of chlorpyrifos (calculated at 7 mg/l air) for 16 exposures over a 21-day interval. There were no effects on growth, appearance, behaviour or plasma and RBC cholinesterase activity (Dow Chemical Co., 1966b). Short-term studies Mallard duck Groups of mallard ducklings (6 - 8 birds/group, 5 days old) were fed chlorpyrifos for five days at dosage levels of 0, 100, 200, 300, 500, 700, 1 000 and 2 000 ppm in the diet. Mortality occurred at all dosage levels (100-2/8; 200-4/7; 300-5/8; 500-6/8; 700 - 2000 - all dead) except in controls (0/7 dead) (Stevenson, 1965). Chickens Groups of chicks (20 chicks/group) were fed chlorpyrifos in the diet for two weeks at dosages of 0, 200, 400 and 800 ppm. Mortality was observed at 400 ppm and plasma cholinesterase depression and reduced weight gain was observed at all levels (Sherman et al., 1967). Groups of chickens (30 per group) were administered chlorpyrifos in the drinking water at dosage levels of 0, 1 ppb, 1 ppm and 100 ppm for 84 days. A higher level (1000 ppm) resulted in clinical signs of cholinergic stimulation. Plasma cholinesterase was depressed at 100 ppm but not at 1 ppm. A 10 ppm group was tested for two weeks with no signs of cholinesterase depression. Growth and behaviour were not affected at 100 ppm (Stevenson and Daering, 1969). Chlorpyrifos added to the drinking water of chicks at dosages of 0, 0.08, 0.32, 1.25, 5, 20, 80, 320 and 1 280 ppm for periods varying from 3 - 5 weeks induced mortality and reduced growth at levels of 80 ppm and above. Whole blood cholinesterase depression was evident only at 80 ppm and above (Brust et al., 1971). Chickens fed chlorpyrifos in the diet at levels of 0, 25, 50 and 100 ppm for four weeks exhibited reduced plasma cholinesterase activity and reduced weight gain at all feeding levels. Calculations of food conversion data show a reduced yield in production as a result of 25 ppm in the diet. No clinical signs of toxicity were observed (Schlinke, 1970). Rat Groups of rats (20 males and 20 females/group) were fed chlorpyrifos in the diet for six months at dosage levels of 0, 0.03, 0.15 and 0.75 mg/kg/day. Cholinesterase depression was noted at 0.75 mg/kg/day in RBC and plasma. Brain cholinesterase activity was unaffected. No effects were noted at any feeding level on mortality, behaviour, growth, haematology or gross or histological examination of tissue (Coulston et al., 1971). Groups of rats (10 males and 10 females/group) were fed chlorpyrifos in the diet for 90 days at levels of 0,10, 30, 100 and 300 ppm. Growth reduction was observed in both males and females at 300 ppm. Increased liver and kidney organ to body-weight ratios were also observed (possibly as a result of reduced body-weight as organ weights were comparable to the controls). Cholinesterase depression was observed at 30 ppm in blood and brain; at 10 ppm, brain cholinesterase was normal although enzyme inhibition was observed in RBC and plasm. No adverse effects were noted on growth, appearance, food consumption, mortality, behaviour, haematology or gross and microscopic pathology at 100 ppm (Beatty and McCollister, 1964). Five groups of rats (10 males and 10 females/group) were fed chlorpyrifos at levels varying from 0.03 - 10.0 mg/kg/day for 28 - 90 days to determine a "no-effect" level on cholinesterase depression. Dosage levels of 0 and 0.3 mg/kg/day were fed for 90 days while dosages of 1, 3 and 10 mg/kg/day were fed for 28 days, stopped for three weeks (during which time the rats were fed control diets) and the rats refed diets containing 0, 0.03 and 0.1 mg/kg/day for a full 90-day study. Behaviour and growth were affected at 3 and 10 mg/kg/day. Cholinesterase was depressed at 0.3 mg/kg/day and above in both male and female rats. The rats in the three highest groups were removed for three weeks during which time cholinesterase activity returned to pretest intervals. On resumption of feeding chlorpyrifos at 0.1 mg/kg/day, slight cholinesterase depression was observed in plasma and erythrocytes. No cholinesterase depression was noted on brain or blood enzymes at 0.03 mg/kg/day (Blackmore, 1968b). Dog Groups of dogs (2 males and 2 females/group; 4 of each sex were controls) were fed chlorpyrifos for 93 days. The dosage levels of feeding were initially 200, 600 and 2 000 ppm in the diet, but because of the appearance of signs of cholinergic poisoning the dosages were changed from 2 000 to 60 ppm after 5 days while the 200 ppm (5.8 mg/kg/day) level was maintained for 45 days. The dogs were then placed on control diets for the remainder of the test. Another group of 2 male and 2 female dogs was added to the study at a dietary level of 200 ppm (3.4 mg/kg/day) for 27 days, removed from the test for 5 days and returned to this dietary level for the final 2 weeks. A decrease in food consumption at the 200 ppm level was accompanied by reduced growth, especially in females. Growth of all dogs was slightly lower than control levels. Increased SGPT values were seen in several dogs at all feeding levels. RBC, plasma and brain cholinesterase values were reduced in all animals fed chlorpyrifos in the diet at a level of 20 ppm (0.8 mg/kg/day) and above. No adverse effects were observed on haematology, BUN, SAP, organ weights and gross or microscopic examination of tissues (Copeland, 1964). Groups of dogs were orally administered chlorpyrifos daily in a gelatin capsule for periods of 32 to 90 days at doses of 0 (2 males and 2 females - 90 days), 0.01 mg/kg/day (2 males and 2 females - 32 days), 0.03, 0.10, 0.30 and 1.00 mg/kg/day (2 males and 2 females/group - 90 days), and a final group (2 males and 2 females) was given 1.00 mg/kg/day from day 0 to 18, control diet from days 19 to 42, 0.03 mg/kg/day from days 43 to 58, control diet from days 59 to 77, 0.01 mg/kg/day from days 78 to 94 and control diet until day 124. Data obtained on appearance, body-weight and food consumption were normal. A dose-dependent cholinesterase depression in plasma was observed to begin at 0.03 mg/kg/day. At this level inhibition was moderate and was more marked as the dietary level increased. Brain cholinesterase was not affected at any level, and RBC cholinesterase was affected at levels of 0.1 mg/kg/day and above. No effects were noted at 0.01 mg/kg/day on any parameters tested over the 32-day administration interval (Blackmore, 1968a). Groups of dogs (7 males and 7 females/group) were fed chlorpyrifos in the diet for up to two years at dose levels of 0, 0.01, 0.03, 0.1, 1.0 and 3.0 mg/kg/day. Inhibition of cholinesterase activity was the only abnormality detected. Inhibition of RBC cholinesterase in males and females was evident at 1.0 and 3.0 mg/kg. Marginal depression of brain cholinesterase was shown at the highest level of feeding. Plasma cholinesterase was depressed at 0.1 mg/kg/day in both males and females. Occasional reduction at 0.03 mg/kg was noted. Studies on the recovery of cholinesterase activity following one year feeding indicate that plasma activity recovered to normal within two weeks while RBC recovery was much slower. No significant effects were noted on mortality, behaviour, food consumption, growth, haematology, blood biochemistry, urinalysis, or gross and microscopic examination of tissues. Gross examination showed an increased liver weight and ratio at 3.0 mg/kg at two years but not at one year. All other tissues were normal (McCollister et al., 1971a). Monkey Groups of Rhesus monkeys (2 males and 2 females - control; 2 males and 1 female - 0.08 or 0.40 mg/kg/day; 2 males and 2 females - 2.0 mg/kg/day) were administered chlorpyrifos orally by gavage daily for six months. Plasma and RBC cholinesterase activity was depressed in those animals that received 2.0 and 0.4 mg chlorpyrifos/kg/day. Plasma cholinesterase activity was slightly depressed and RBC cholinesterase activity was not depressed in animals that received 0.08 mg/kg/day. Brain cholinesterase activity was not affected at any level. No significant effects were noted on behaviour, growth, haematology, clinical chemistry or gross or histological examination of tissues. The histological examination included electron microscopy of liver and kidneys and histochemical delineation of lysosomal distribution in these two organs (Coulston et al., 1971). Long-term studies Rats Groups of rats (57 males and 57 females/group) were fed dietary levels of chlorpyrifos of 0, 0.01, 0.03, 0.1, 1.0 and 3.0 mg/kg/day for two years. Blood cholinesterase activity, primarily in plasma, was depressed in females at 0.1 mg/kg and above. Brain cholinesterase was inhibited at 3.0 mg/kg and slightly depressed at 1.0 mg/kg. Chlorpyrifos at all dosage levels had no significant effect on behaviour, appearance, growth, mortality, haematology, urinalysis, clinical biochemistry, gross or histopathology of tissues and organs or the incidence of neoplasms (McCollister et al., 1971b). OBSERVATIONS IN MAN Groups of men (4 men/group) were administered chlorpyrifos by tablet daily for 27 days (0.014 mg/kg/day), 20 days (0.03 mg/kg/day), 9 days (0.10 mg/kg/day) and 48 days (control). No effects were noted on behaviour, haematology, urinalysis or biochemical determinations in blood. Blood plasma cholinesterase was depressed at the highest dosage and this regimen was discontinued. Recovery was complete in four weeks. At 0.03 mg/kg/day a statistically non-significant, although suggestive, depression of plasma cholinesterase was noted. No effects were noted at 0.014 mg/kg on plasma cholinesterase. RBC activity was unaffected at any level of chlorpyrifos. The level of 0.03 mg/kg/day appears to be the minimal threshold response level in humans to chlorpyrifos based on plasma cholinesterase (Coulston et al., 1972). COMMENT The compound is rapidly absorbed and metabolized by mammals, the major metabolite being a chlorinated pyridonal hydrolysis product of low mammalian toxicity. There is no evidence of neurotoxicity or cataractogenicity. Potentiation has been demonstrated only with malathion. Reproduction studies and examination of offspring for teratogenic abnormalities did not reveal adverse effects other than an increase in neonatal mortality at 1 mg/kg/day. Chlorpyrifos is an active cholinesterase inhibitor, inhibiting plasma cholinesterase to a far greater degree than other cholinesterases. One short-term study in rats indicated an increase in sensitivity to plasma cholinesterase depression following withdrawal after initial treatment. This study needs confirmation. No-effect levels have been demonstrated in dog, rat and man. TOXICOLOGICAL EVALUATION Level causing no toxicological effect Rat: 0.03 mg/kg/day Dog: 0.01 mg/kg/day Man: 0.014 mg/kg/day orally for 1 month ESTIMATE OF ACCEPTABLE DAILY INTAKE FOR MAN 0 - 0.0015 mg/kg body-weight RESIDUES IN FOOD AND THEIR EVALUATION USE PATTERN In animals Chlorpyrifos is used to control various species of fever ticks (Boophilus sp.), ear ticks, lice and horn flies on beef cattle and non-lactating dairy cattle, by use of emulsifiable liquid formulations in water varying from 0.025 to 0.125% applied as a spray or dip. Treatment for all but ear ticks is limited to six applications at 21-day intervals, and not within two weeks of slaughter. Sheep dipped or sprayed with wettable powder or emulsifiable formulations of chlorpyrifos are protected from blowfly, ticks, body lice and sheep keds. A minimum interval of seven days is required between treatment and slaughter. Turkey chiggers are controlled through the use of wettable powder formulations sprayed on soil in turkey pens at the rate of 4 lb/acre chlorpyrifos, before the turkeys are introduced into the pens. Treatment is limited to two applications, not within seven days of slaughter. In plants Soil treatments 1. Europe, Near East and South Africa Chlorpyrifos is used to control soil pests such as wireworms, white grubs, mole crickets, Maulwurf's Grill, garden symphylans, seed corn maggots, cereal maggots, beet root weevils, the Atomaria, Blaniulus, Tipula and cutworms. The chemical is applied by means of broadcast row or furrow treatments and is varied in concentrations from 300 to 4 000 g/ha a.i. as wettable powder, emulsifiable or granular formulations before and after planting, previous to harvest. The crops involved are maize, spring cereals, beans, onions, tobacco, turnips, bulb flowers and carrots. 2. Canada Chlorpyrifos is used to control soil insect pests of lettuce, onions and carrots before and after planting at dosages of 1 to 2 lb/acre applied as needed, but with the indicated 28- to 60-day withdrawal period before harvest. Tobacco plants are treated in soil water, preplant and by soil and foliar application. Foliar treatments 1. Europe, Near East and South Africa Chlorpyrifos is used to control insect pests of rape, cereals, potatoes, fodder crops, beans, peas and deciduous fruits such as apples and pears. 2. Japan, Thailand, Philippines, India and Malaysia Chlorpyrifos is used to control insect pests of apples, pears, rice, tobacco, Chinese cabbage and kale. RESIDUES RESULTING FROM SUPERVISED TRIALS In animals Cattle Considerable work has been done in U.S.A. on the analysis of cattle tissues for residues of chlorpyrifos as well as its oxygen analogue and pyridinol moiety. Typical distribution of residues in cattle dipped in chlorpyrifos is shown in Table 3 (Claborn et al., 1970a). In this study, 57 beef cattle were dipped one to six times in a 0.025% emulsion of chlorpyrifos at intervals of 21 days. Animals were removed from the dipping schedule after each dip, except the fifth, for post-treatment slaughter at various time intervals. Fat muscle, kidney and liver tissue samples were collected and analysed for residues of chlorpyrifos, its oxygen analogue, and the 3,5,6-trichloro-2-pyridinol moiety. Residues of chlorpyrifos were predominately in the fatty tissues, and residues of 3,5,6-trichloro-2-pyridinol were in the kidney and liver tissues. No residue of the oxygen analogue was detected in any tissue. Residues of the hydrolytic pyridinol were not detectable in muscle tissue or in kidney and liver tissues of cattle, except at low levels for the first 1 - 2 weeks after the last dip, even after six dippings. It should be noted that this experiment began in February, so the animals had their winter coat of hair through the first three dippings and were shedding it at approximately the fourth dipping period. The heavy winter hair coat carries more insecticide out of the vats than the summer coat, as evidenced by the higher residues after the second and third dippings and the drop in residues after the fourth dipping. The effect of the winter coat is also reflected by the higher residues after the first dip when compared to levels of residues after a single dip (in the same concentration or higher concentration - 0.05% - of chlorpyrifos) in the animals reported by Mann, 1966; Claborn et al., 1968a,b; and Dishburger et al., 1966a,b. Since the season for ticks extends from approximately April or May to mid-September, animals would be expected to shed their winter coat after the first or second dipping when treated according to label instructions. Therefore, in practice residue levels as high as those found in the test animals would not be expected after the third dipping, since these animals would have been dipped three times before shedding their winter coat of hair. In any case, it is believed that this experiment represents the highest levels of chlorpyrifos that would ever be encountered in the use of 0.025% chlorpyrifos as a dip. In an early test, cattle were sprayed once with a 0.5% emulsion of chlorpyrifos (20 times the label recommendation). Residues of the compound in the fat were 1.3 ppm (maximum) at 20 days post-treatment and had declined to 0.08 ppm (maximum) at 30 days post-treatment (Dishburger and Rice, 1965). In cattle treated with an emulsion containing 0.05% chlorpyrifos (twice the label recommendation), the average amounts of residue in the fat one week after a single-dip application were about twice those resulting from a single-spray application. The maximum residues were well below 1 ppm (0.209 for spray and 0.525 for dip application) at the first sampling period at seven days post-treatment. Approximately two weeks were required for residues to be reduced to 1 ppm in the fat of animals given multiple treatments (3 dippings at 2-week intervals) at this level (Mann, 1966; Claborn et al., 1968a,b). TABLE 3 Chlorpyrifos residues in cattle in the United States1 Mean residues of Weeks Mean residues of chlorpyrifos2 3,5,6-trichloro-2-pyridinol3 after last fat muscle kidney kidney liver dipping (ppm) (ppm) 1 dipping 1 0.672 0.004 0.003 0.12 0.12 2 0.142 0.002 0 <0.05 <0.05 3 0.083 0.001 0 0 0 4 0.038 0 0 0 0 2 dippings4 1 1.245 0.011 0.008 0.30 0.26 3 dippings4 1 1.589 0.028 0.008 0.24 0.29 2 0.085 0.011 0.001 0.05 0.05 4 0.158 0 0 0 0 6 0.038 0 0 0 0 8 0.041 0 0 0 0 10 0.001 0 0 0 0 4 dippings4 1 0.693 0.011 0.001 0.10 0.12 6 dippings4 1 0.744 0.015 0.003 0.12 0.10 2 0.141 0.001 0 0 0 4 0.069 0 0 0 0 6 0.018 0 0 0 0 8 0.007 0 0 0 0 10 0 0 0 0 0 1 Residues of chlorpyrifos and 3,5,6-trichloro-2-pyridinol in tissues of beef cattle dipped in an emulsion containing 0.025% chlorpyrifos. Residue levels corrected for % recovery from tissues. 2 No chlorpyrifos found in liver (0 - <0.002 ppm) or oxygen analogue (0 - <0.005 ppm) in any tissue analysed (Claborn, 1970). 3 No 3,5,6-trichloro-2-pyridinol found in fat or muscle tissues (0 - <0.05 ppm) (Dishburger and McKellar, 1971). 4 Dipping schedule was at 21 day intervals. The oxygen analogue was found only in samples of fat containing the higher residues of chlorpyrifos (approximately 2 ppm); it amounted to about 1% of the residue. No increase occurred in the amount of the oxygen analogue as the chlorpyrifos was eliminated, and no residues of the analogue were detected in the muscle, brain or spleen tissues of the animals that had the highest residues of chlorpyrifos in the fat (Claborn et al., 1968a). Data from Mann (1966), Claborn et al. (1968a) and Dishburger et al. (1966a,b) indicated that the biological half-life of chlorpyrifos in tissues of cattle following a single treatment with DURSBAN insecticide containing 0.025% and 0.05% chlorpyrifos is approximately 4 - 6 days. Data from Mann (1966) and Claborn et al. (1968a,b) showing multiple dippings in DURSBAN insecticide containing 0.025% and 0.05% chlorpyrifos, further establish the consistency of the degradation ratio following the last treatment. The data of Claborn et al. (1968b) indicate that the biological half-life of chlorpyrifos in tissues of cattle treated at three-week intervals for as long as six weeks with DURSBAN 24E containing 0.025% chlorpyrifos is approximately seven days. In summary, the experiments illustrate the consistency of a 4 - 7 day biological half-life for chlorpyrifos in fat. Considerable work has been carried out in Australia on the determination of residues from the vat dip and spray application of chlorpyrifos to cattle (Hall, 1969; Snelson, 1969b). Results shown in table 4 indicate that residues of chlorpyrifos in omental fat are in the same order of magnitude as those for United States, considering the fact that most tests were at double the concentration (0.05%) in Australia. Snelson (1969a) studied the disappearance rate of chlorpyrifos in butterfat in milk after a single dip application of chlorpyrifos at 0.025% to milk cows. Residue declined from 1.1 ppm after 1 day to 0.01 ppm in 9 days, and a non-detectable level at 15 days (see Table 5). Sheep Sheep dipped in an 0.05% chlorpyrifos emulsion showed inconsistent levels of residues of chlorpyrifos with no apparent change in residue levels (0.081 - 0.179 ppm) from 3 - 28 days after application. No detectable levels (<0.005 ppm) were seen in muscle, liver or kidney tissues at any time (Dow Chemical Co., 1967). TABLE 4 Chlorpyrifos residues in cattle in Australia Chlorpyrifos Days residues in Chlorpyrifos Application between last No. of omental fat Reference % type no. treatment and cattle (mean values slaughter or range, ppm) 0.025 Vat dip 3 1 0.2 Watts, 1968 4 0.96 7 0.25 1 1 3 0.11 - 0.22 Ibid. 1 1,4,8,15 0.29 - 2.18 Ibid. 3,6 1,4,8,15 0.05 - 1.19 Ibid. 0.05 Vat dip 1 1,4,8 0.41 - 0.48 Ibid. 3 1 1.17 Ibid. 4 0.98 6 1 1.75 Ibid. 8 1.36 15 1.11 22 0.6 29 0.38 0.05 Vat dip 2 0 3 1.46 - 2.26 Hall, 1969 1 2 3 2.4 - 3.87 1 2 3 1.23 - 3.09 1 2 3 0.98 - 2.54 1 1 3 0.42 - 0.79 3 1 4 1.32 - 3.80 0.05 Dip 10 5? 12 0.70 - 7.131 Snelson, 1969b 0.05 Spray 30 1 3 1.6 - 4.2 Ibid. 5 3 0.34 - 1.1 9 2 0.1 - 0.15 13 1 0.06 28 1 0.01 1 Omental and/or perirenal fat. TABLE 5 Chlorpyrifos residues in milk of cows in Australia Days between Chlorpyrifos1 last treatment residue in butterfat and sampling (ppm avg. 4 cows) Reference 1 1.1 Snelson, 1969a 2 0.96 3 0.57 4 0.14 5 0.08 6 0.05 9 0.01 15 N.D.2 1 One dip application containing 0.025% chlorpyrifos. 2 N.D. = not detectable Turkeys and chickens Claborn et al. (1970b) determined residues of chlorpyrifos in the body tissues of turkeys confined in pens containing soil treated with 4 or 8 lb/acre of chlorpyrifos (in a 25% wettable powder formulation) for control of the chigger, Neoschongastia americana. The higher concentration caused maximum residues of 0.157 and 0.066 ppm of chlorpyrifos in the skin and fat, respectively, one week post-treatment, that decreased to less than 0.001 ppm after six weeks. No significant residues were found in the other tissues (thigh, breast, liver). The oxygen analogue of chlorpyrifos (diethyl 3,5,6-trichloro-2-pyridyl phosphate) was not detected (<0.005 ppm) in any tissues. In a study conducted by Mann and Ivey (1971), turkeys were maintained on soils treated two times, at an interval of 28 days, with 4 lb chlorpyrifos/acre each time. Two or three birds were necropsied at weekly intervals for six weeks after the first application and for eight weeks after the second application. Tissue samples of fat, skin, muscle, liver and kidney were analysed for residues of chlorpyrifos, the oxygen analogue and the pyridinol moiety (3,5,6-trichloro-2-pyridinol). Lowlevel residues of chlorpyrifos were found in fat (0.022 - 0.005 ppm), skin (0.033 - 0.152 ppm) and kidney (<0.002 - 0.005 ppm) of birds after one week of exposure to the first application of compound to the soil. Fat only was analysed for the remainder of the test for each application. The residues in fat were reduced to <0.002 ppm (the sensitivity of the method) six weeks after the first application and eight weeks after the second application. No oxygen analogue was detected in any of the tissues. Small residues of the pyridinol moiety were found in the skin of one hen (<0.003 - 0.007 ppm), the liver of one tom (<0.01 - 0.025 ppm) and the kidney of one tom (<0.01 - 0.098 ppm) after one week of exposure to the first application; and in the skin (<0.003 - 0.003 ppm), liver (<0.01 - 0.078 ppm) and kidney (<0.01 - 0.165 ppm) in one hen after one week of exposure to the second application of compound to the soil. Hunt et al. (1969) developed a new rapid gas chromatographic method for analyses of chlorpyrifos in turkey and chicken tissues. The residues reported are from the same experiments as the toxicological results reported by Schlinke (1970) in chickens and Schlinke et al. (1969) in turkeys. The tests consisted of dietary and contact tests with chlorpyrifos for four weeks. Residue analyses of muscle, liver, heart, brain, gizzard, skin and fat tissues were taken after 28 days of treatment. Only skin and fat tissues showed detectable amounts of chlorpyrifos. In chickens fed 100, 50 and 25 ppm in the diet, the highest tissue residue was 0.87 ppm chlorpyrifos in fat. In turkeys fed 100 and 50 ppm in the diet, the highest residue was 0.34 in fat. Dishburger et al. (1969b) determined the presence of chlorpyrifos residues in skin, fat, muscle and liver tissues of turkeys which had been confined to soil treated with 4.5 lb chlorpyrifos/acre as emulsifiable, granular, or wettable powder formulations. Muscle and liver tissues contained undetectable amounts of chlorpyrifos in all tests. Skin and fat tissues contained <0.1 ppm at all times, but were higher at seven days post-treatment than any of the other weekly samplings up through 28 days post-treatment. In plants Residues found in experiments with plants in United States Chlorpyrifos has been applied to various species of plants and generally found to be lacking in residual insecticidal properties on plant-feeding insects. The residue determinations reported here actually never exceeded 28 ppm at the dosages tested (up to 1 lb/acre) and soon diminished, exhibiting a half-life of 1 - 2 days, thus confirming the reason for the observed lack of residual effectiveness on plant-feeding insects (see Table 6). There was no evidence of residue buildup on plants from repeated application of chlorpyrifos. TABLE 6 Chlorpyrifos residues found in experiments in plants in United States Dosage Application Chlorpyrifos chlorpyrifos (no.) (no. of days Crop residues Reference (lb/A) since last) (ppm) 1.0 1 0 Bermuda grass 13.2 Leuck et al., 1968 14 0.37 1.0 1 0 Maize 5.60 Ibid. 14 0.42 0.05 8 0 Pasture grass 0.7 - 8.2 Winterlin et al., 7 <0.1 - 0.2 1968 0.05 1 0 Grass 0.11 - 8.1 Ibid. 3 0.012 - 1.8 0.025 1 0 Rice 0.09 - 0.64 Ibid. 6-14 <0.005 - 0.07 1.0 1 0 Various wild 26.9 Hurlbert et al., 1 plants 27.4 1970 7 1.1 14 0.1 0.025 4 0 Rice 0.07 - 0.21 Dishburger et al., 9 <0.01 1969c. 1.0 1 0 Maize silage 15.1 Johnson et al., 1969 1 4.4 0.25 1 0 Maize silage 2.5 Ibid. 1 1.1 Leuck et al. (1968) studied the persistence of chlorpyrifos and its oxygen analogue on field plots of coastal Bermuda grass and maize forage using the method of Bowman and Beroza (1967a) with a minimum sensitivity of 0.002 - 0.01 ppm. Chlorpyrifos was applied at the rate of 1 lb/acre. The oxygen analogue constituted a very low residue and was only a small proportion of the total insecticide. Chlorpyrifos residues varied from about 6 - 13 ppm at the time of application to 4 - 8 ppm after 1 day, to around 1 ppm after 7 days and to nearly negligible levels after 21 days. The residue half-life of chlorpyrifos appeared to be in the order of 2 - 3 days on Bermuda grass and maize plants. Winterlin et al. (1968) studied the residues of chlorpyrifos on rice and pasture grass and environment when applied experimentally for the control of mosquitoes around Colusa, California. They used the method of Bowman and Beroza (1967a) i.e. gas chromatography with a flame ionization detector, which is sensitive to 0.005 ppm chlorpyrifos in rice and 0.01 ppm in pasture grass, and found residues as high as 8.11 ppm chlorpyrifos immediately after application of 0.05 lb chlorpyrifos/acre. Residues decreased rapidly after application to below 1 ppm. Myers et al. (1968) reported on residues resulting from application of chlorpyrifos sprayed by aircraft at the rate of 0.1 lb/acre, eight times, over a period of five months on irrigated pastures. Samples of grass analysed for the insecticide 1 - 3 hours after each spray application varied from 0.4 - 8.2 ppm, and after seven days varied from <0.1 - 0.2 ppm, using an analytical method sensitive to 0.1 ppm chlorpyrifos. No evidence of accumulated residues of chlorpyrifos were detected as the result of multiple applications to grass with chlorpyrifos. Hurlbert et al. (1970) reported on the residues of chlorpyrifos on watergrass, sprangletop and smartweed on the dikes surrounding ponds, resulting from hand-sprayed applications of chlorpyrifos emulsifiable insecticide. Dosages used were 1, 0.1, 0.05, and 0.01 lb/acre diluted in a constant spray volume of 8 gal/acre. Under these conditions residues over 1 ppm occurred only at 1 lb/acre after 4 h, 1 day, and 7 days, but not 14 days; and at 0.1 lb/acre after 4 h. Residues determined at other dosages and post-application times were less than 1 ppm. The half-life of the foliar residues of chlorpyrifos was in the order of 1 - 3 days. Dosages of 0.025 lb chlorpyrifos were applied aerially four times, at monthly intervals, to rice fields. Residues of chlorpyrifos in raw (rough) rice grain ranged from 0.07 to 0.21 ppm at 0 day, following the last of the four applications. No residues of the oxygen analogue of chlorpyrifos were detected, nor did detectable chlorpyrifos residues persist more than 9 days after application (Dishburger et al., 1969c). Johnson et al. (1969) treated maize plants in the field with 0.25, 0.5 and 1.0 lb chlorpyrifos/acre. The initial residues in maize and silage were 2.5, 3.9 and 15.1 ppm, respectively (on an as-collected basis). After one day the losses of residues were 56, 40 and 71%, respectively. This represents a half-life of about one day. One day after treatment the maize and silage were enclosed in a silage bin where chlorpyrifos residues declined slowly over a period of seven weeks. Dishburger et al. (1969a) analysed samples of tobacco leaves from soil and foliage treated with 0.5, 1.0 and 1.5 lb chlorpyrifos/acre. No residues (<0.01 ppm) were detected on the samples harvested (2 months or more post-application). Residues found in experiments with plants in Canada Residue data from experiments with onions, lettuce, carrots, and tobacco are shown in Tables 7 and 8 using the analytical method of Braun (1971) for chlorpyrifos and McKellar (1971c) for the pyridinol. TABLE 7 Residues of chlorpyrifos from treatments of lettuce, onions and carrots1 Application Residues on crop (ppm) (lb/acre) (No.)2 lettuce onions carrots3 0.5 2 <0.001 0.001 <0.001 0.005 0.003 0.003 (54)4 (119) (138) 3 0.001 0.002 0.012 0.020 0.003 0.020 (31) (104) (123) 1.0 2 <0.001 0.002 0.012 0.008 0.005 0.034 (54) (118) (138) 3 0.002 0.002 0.006 0.009 0.005 0.018 (31) (104) (123) 2.0 2 0.003 0.008 0.015 0.029 0.013 0 046 (54) (119) (138) 2.0 3 0.008 0.003 0.019 0.034 0.005 0.033 (31) (104) (123) 1 McEwen and Frank, 1970. 2 First application was preplant soil treatment; second and third soil and foliar treatments (Canada). 3 3,5,6-trichloro-2-pyridinol analysed for but not found at detection limit of 0.005 ppm. 4 Days after chlorpyrifos withdrawal shown in (). TABLE 8 Residues of chlorpyrifos from treatments of lettuce, onions, carrots and celery Withdrawal Applications1 Residues range Crop (days) (no.) (ppm) References Onions 57 1 0.001 - 0.003 McEwen, 1970 19 4 <0.0012 Harris, 1971 Celery 57 1 N.D.3 - 0.016 McEwen, 1970 Carrots 57 1 0.001 - 0.015 Ibid. Lettuce 27 1 N.D. Ibid. 1 Treatments all post-planting, 2 lb/acre (Canada). 2 No oxygen analogue of chlorpyrifos detected at 0.01 ppm sensitivity level. 3 N.D. = <0.001 ppm. Residues found in experiments with plants in Europe and the Near East Residue data from experiments with cauliflower, red cabbage, eggplant, beans, peppers, lettuce, mushrooms, carrots, potatoes, sugar beet, corn, cotton, tomatoes, grapes, pears and apples are shown in Tables 9 and 10. These tests include soil and foliar applications. Residues found in experiments with plants in Japan, Thailand, Malaysia, and the Philippines Residue data from experiments with Chinese cabbage and kale, rice and apples are shown in Tables 11, 12 and 13. TABLE 9 Residues of Chlorpyrifos in Food Crops, Europe/Near East Trials COUNTRY YEAR FORMULATION APPLICATION DOSAGE1 INTERVAL2 RESIDUES REFERENCES and CROP TYPE NO. (ppm) FRANCE Potatoes 1970 Granules Soil 1 1 500 g/ha 5 mo <0.005 Graf and Engeler, 5% active insecticide 1 3 500 g/ha 5 mo <0.005 1971a W. GERMANY Sugar beets 1971 Dursban EC Broadcast 1 1.15 kg/ha 112 d <0.13 Scheuerer & Hahn, 40.8% active Soil insect. 118 d 1972 Maize 1971 Dursban 4 EC Soil insect. 1 0.95 kg/ha 123 d <0.13 Ibid. 40.8% active Broadcast HOLLAND Cauliflower Strewpowder Soil insect. 1 120 mg 8 wk 0.002 Red cabbage (1) 1969 4% active 11 wk <0.002 Wit, 1970 Red cabbage (2) 85 wk <0.002 Lettuce (under 1970 3.2% active Soil insect. 1 1 900 g/ha 9 wk <0.01 Wit, 1971 glass) granule Mushrooms 1971 25% wettable Spray 3 5 000 g/ha 3 wk 0.01 Olthof, 1971a powder 3 wk 0.03 Carrots 1971 Dursban 25W Soil insect. 1 6 000 g/ha 3-4 mo 0.14 Olthof, 1971b 1 6 000 g/ha 3-4 mo 0.28 ISRAEL Cotton 1970 Dursban E.C. Spray 6 730 g/ha 15 d <0.05 Graf, 1970a 40.8% active Apples 1970 Dursban E.C. Spray 1 2 400 g/ha 7 d 0.54 Graf & 40% active 14 d 0.20 Engeler, 1970a Tomatoes 1970 Dursban E.C. Spray 1 720 g/ha 1 d 0.24 Aharonson 40.8% active 4 d 0.16 et al., 1969 8 d 0.12 16 d 0.04 1 1 440 g/ha 1 d 0.45 TABLE 9 (Cont'd.) COUNTRY YEAR FORMULATION APPLICATION DOSAGE1 INTERVAL2 RESIDUES REFERENCES and CROP TYPE NO. (ppm) 4 d - 8 d 0.18 16 d 0.10 1971 Dursban E.C. Spray 1 1 440 g/ha 1 d 0.27 Graf & Engeler, 40.8% active 5 d 0.25 1971b Apples 1971 Dursban E.C. Spray 1 1 440 g/ha 1 d 0.57 Graf & Engeler, 40.8% active 7 d 0.23 1971c 14 d 0.14 Spray 1 2 880 g/ha 1 d 0.52 7 d 0.28 14 d 0.24 Dursban Spray 1 1 440 g/ha 1 d 0.13 40.8% active 7 d 0.11 Spray 1 2 880 g/ha 1 d 0.29 Ibid. 7 d 0.24 ITALY Sugar beets 1970 Dursban 5% Soil insect. 1 2 500 g/ha 3-6 mo <0.004 Foschi, 1971 Maize 1970 Granule Soil insect. 1 2 500 g/ha 3-6 mo <0.004 Lettuce 1971 Dursban E.C. Spray 1 500 g/ha 14 d 0.05 SIAPA, 1971 22.5% 1 1 000 g/ha 14 d 0.07 Apples 1970 Dursban E.C. Spray 1 816 g/ha 15 d 0.16 Graf, 1970b 40.8% active 29 d 0.10 1 1 224 g/ha 15 d 0.25 29 d 0.16 Pears 1970 Dursban E.C. Spray 1 816 g/ha 15 d 0.22 Graf, 1970c 40.8% active 29 d 0.14 1 1 224 g/ha 15 d 0.22 29 d 0.27 TABLE 9 (Cont'd.) COUNTRY YEAR FORMULATION APPLICATION DOSAGE1 INTERVAL2 RESIDUES REFERENCES and CROP TYPE NO. (ppm) Turkey Eggplant 1972 Dursban 4 Spray 3 0.96 kg/ha 0 d 0.26-0.40 Hollick & E.C. 7 d 0.02-0.03 Collison, 1972 14 d 0.02-0.02 Beans 1972 Dursban 4 Spray 2 0.96 kg/ha 0 d 0.65-1.20 Ibid. E.C. 7 d 0.08 14 d <0.04 Tomatoes 1970 Dursban E.C. Spray 1 347 g/ha 3 h 1.3 Graf & Engeler, 40.8% active 5 d 0.26 1970b Peppers (under 1972 Dursban 4 Spray 3 0.96 kg/ha 0 d 2.00-5.20 Hollick & glass) E.C. 7 d 0.04-0.10 Collison, 1972 14 d 0.13-0.25 UNITED KINGDOM Raspberries 1971 Dursban E.C. Spray 1 0.1% 0 d 1.40 Burrows et al., 40.8% active 5 d 0.32 1971d 2 0.5% 10 d 0.14 18 d 0.18 Tomatoes 1971 Dursban E.C. Spray 1 0 d 1.26 Burrows et al., 40.8% active 3 d 0.10 1971e 7 d 0.08 14 d 0.01 28 d 0.01 1 All dosages given in a.i. 2 Last application to harvest 3 Limit of detection TABLE 10 RESIDUES OF CELORPYRIPOS AND PYRIDINOL IN FOOD CROPS, EUROPE/NEAR EAST TRIALS CROP, SAMPLE DOSAGE1 TIME BETWEEN RESIDUES RESIDUES REFERENCES COUNTRY NO. LAST TREATMENT CHLORPYRIFOS 3,5,6-TRICHLORO-2-PYRIDINOL (ppm) & YEAR AND HARVEST (ppm) TURKEY 1971 Tomatoes 1 A 825 g/ha 3 hours 1.3* <0.05 2 A " 3 hours <0.05 4 A " 3 hours 0.05 Barrows & Mullin, 3 B " 5 days 0.05 1971b 5 B " 5 days 0.26 <0.05 6 B " 5 days <0.05 ENGLAND 1971 Tomatoes 720 g/ha 0 day 1.28 0.14 1.24 0.12 3 days 0.14 0.10 0.05 0.10 Burrows et al 7 days 0.08 0.08 1971e 0.08 0.08 14 days 0.01 0.12 0.01 0.10 28 days <0.01 0.10 <0.01 0.16 ITALY 1971 Apples 1 816 g/ha 29 days 0.10 <0.05 Burrows & Mullin, 4 1 224 g/ha 15 days 0.25 <0.05 1971b Grapes 11 816 g/ha 29 days 0.10 <0.05 13 816 g/ha 15 days 1.10 0.06 Ibid. 12 1 224 g/ha 29 days 0.22 <0.05 14 1 224 g/ha 15 days 1.50 0.29 * Composite samples 1 All dosages given in a.i. TABLE 11 CHLORPYRIPHOS RESIDUES ON CROPS IN JAPAN1 CROP FORMULATION CHLORPYRIFOS APPLIC. DAYS BETWEEN RESIDUE2 & and DILUTION IN SPRAY (no.) LAST SPRAY (ppm) YEAR (ppm) AND HARVEST Chinese cabbage 1967 40E 267 1 4 0.13-0.23 1100 l/ha 1969 40E 267 2 8 0.02-0.09 1000 l/ha 2 16 0.43-0.44 " 267 4 8 0.16-0.22 4 16 0.62-0.86 1969 40E 267 2 7 0.02-0.06 1500 l/ha 2 14 0.06-0.08 " 267 4 7 0.08-0.14 4 14 0.06-0.12 Apples Rall's Janet 1967 25W 250 4 66 N.D.3 Johnathan 1967 25W 250 5 42 0.01-0.05 1969 25W 250 3 30 0.05-0.07 30 l/tree 45 0.05-0.06 1969 " 250 2 17 0.03-0.07 28 0.02-0.05 46 0.02-0.03 TABLE 11 (Cont'd.) CROP FORMULATION CHLORPYRIFOS APPLIC. DAYS BETWEEN RESIDUE2 & and DILUTION IN SPRAY (no.) LAST SPRAY (ppm) YEAR (ppm) AND HARVEST 25W 250 3 7 0.03-0.09 30 l/tree 14 0.05-0.17 31 0.08-0.16 45 0.03-0.05 Aomori 1969 25W 250 2 7 0.10 14 0.12-0.14 21 0.08-0.03 1 Hamaguchi, 1970 2 Range of averages for two laboratories 3 N.D. = not detectable (<0.005 ppm) TABLE 12 CHLORPYRIFOS RESIDUES ON CROPS IN THAILAND AND MALAYSIA CROP FORMULATION APPLIC. DAYS BETWEEN TESTS RESIDUE1 REFERENCES & DILUTION (no.) LAST SPRAY (no.) (ppm) & HARVEST Chinese 25W 6 5 6 N. D.2 - 0.023 Tucker & Zielinski, cabbage 1972a Chinese 187 ppm(a.i.) " " " 0.10 - 0.243 Ibid., 1972b kale " " " " 0.09 - 0.763 Chinese 25W " " " N.D.3 Ibid., 1972a cabbage Chinese 375 ppm(a.i.) " " " 0.38 - 0.753 Ibid., 1972b kale " " " " 0.35 - 0.763 Chinese 25W 16 2 3 0.07 - 0.104 Ibid., 1972, kale 3 oz a.i./A " " " 0.01 - 0.35 25W " " " 0.13 - 0.244 4 oz a.i./A " " " 0.01 - 0.035 Tobacco 25W 10 7 4 0.03 - 0.053 Ibid., 1972d 187 ppm (a.i.) 25W " " " 0.21 - 0.383 375 ppm (a.i.) 1 Two analytical methods used in analyses 2 N.D. = not detectable (<0.01 ppm) 3 "Usable leaves" from Thailand 4 Leaf only from Malaysia 5 Stem only from Malaysia TABLE 13 Chlorpyrifos residues on rice in the Philippines1 Chlorpyrifos Residue applied (kg/ha, Tests (range of Part analysed Formulation 3 applications) (no.) averages, ppm) Grain E.C. 0.10 4 N.D.3 0.15 4 N.D. - 0.008 0.20 3 N.D. - 0.005 0.25 2 0.008 Granule 0.10 3 N.D. Grain and E.C. 0.10 4 N.D. - 0.007 straw2 0.15 4 N.D. - 0.006 0.20 4 0.005 - 0.008 0.25 2 0.008 Granule 0.10 3 <0.005 1 Tucker, 1971. 2 Very little straw, mostly grain. 3 N.D = not detectable (<0.005 ppm) FATE OF RESIDUES General comments The fate of chlorpyrifos residues and its metabolites in different species of animals are discussed in the section on "Biochemical aspects". In animals Cow Chlorpyrifos was not found in the urine and milk from a cow fed 5 ppm (based on a daily ration of 50 lb) of the compound for four days. The compound in absolute ethanol was mixed thoroughly with the grain. A compound characterized by retention time as chlorpyrifos was found in faecal samples taken on the last three days during which the insecticide was fed and the first day after, and represented 1.7% of the insecticide fed. Two metabolites were excreted in the urine which, following methylation, had retention times identical to the methyl esters of diethylthiophosphate and diethyl phosphate. They represented 35.9 and 26.8%, respectively, of the total insecticide fed on an equivalent basis (Gutenmann et al., 1968). Fish Waters treated with 50 and 300 ppb of ring-labelled 14C chlorpyrifos and containing goldfish were analysed for the types of metabolites found in each. Metabolites I, III, IIIa and IIIb, shown in Figure 1, as well as chlorpyrifos were found in fish, and metabolites I, IIIa and IIIb were found in the water (Smith et al., 1966). The major metabolic product in fish is 3,5,6-trichloro-2-pyridinol which appears to be slowly broken down by dehalogenation and cleavage of the ring (Smith et al., 1966). Some of the derivatives of 3,5,6-trichloro-2-pyridinol may be those shown in Smith (1968) which consist of dechlorination forming a series of transient pyridine diols, triols and tetrols (Ia), followed by a series of transient ring diketones (Ib), followed by a breakage of the ring to elemental fragments (carbon dioxide and possibly aliphatic amines). Cholinesterase inhibition properties of metabolites The house fly head cholinesterase activity of chlorpyrifos, and metabolites I, II, III, IIIa and IIIb shown in Figure 1, were tested for comparative purposes by Smith et al. 1966). Only the phosphate derivative of chlorpyrifos (metabolite III) shows significant inhibition and is the compound most likely to be responsible for the cholinesterase inhibition found in mammals and other animal organisms (Smith et al., 1966). In plants Studies on the absorption, translocation and metabolism of 14C and 36Cl labelled chlorpyrifos in plants show that for all practical purposes uptake of the chemical or its degradation products into plants does not occur, either through the foliage or through the roots (Smith et al., 1967a, 1967c). When radioactive chlorpyrifos was applied to single bean or corn leaves, about 75 to 80% of the chemical was lost by vapourization from the leaf surface within the first 48 h following treatment. The chlorpyrifos remaining on the leaf was rapidly degraded (>95% in 5 days) and a trace of the resulting breakdown products (representing <2% of the applied dose of chlorpyrifos) absorbed into and translocated within the plant. The identity of the degradation products was not totally elucidated; however, a variety of hydrolysis products appeared to be formed, mainly the 3,5,6-trichloro-2-pyridinol (I), ethyl-3,5,6-trichloro-2-pyridyl phoshate (IIIa) and 3,5,6-trichloro-2-pyridyl phosphate (IIIb) shown in figure 1 (Smith et al., 1967a).
Nutrient culture experiments have shown uptake of chlorpyrifos by plant roots to be essentially nil. Bean plants grown in nutrient solution containing 10 mg (50 ppm) of 14C chlorpyrifos absorbed and translocated to the plant tops less than 0.07% of the applied dose during a 144 h exposure period. Based on the weight of plant tissue analysed, this represented a residue <0.002 ppm of radioactive compounds (expressed as chlorpyrifos). In this same experiment, sizable amounts of chlorpyrifos were sorbed on the outside of the plant roots, reaching 5 mg (50%) in 72 h. The negligible amounts of radioactivity taken into the plants in this and other tests have made it difficult to determine the nature of the compounds involved. Only the 3,5,6-trichloro-2-pyridonol was identified. This compound, the principal hydrolysis product of chlorpyrifos in soil, may or may not be taken up by the plant depending on pH of the soil medium. The free pyridinol, which is the predominant form of the compound below pH 6.0, is essentially insoluble in water. Soil and nutrient culture experiments have shown its uptake by plants to be insignificant. For example, bean plants grown in nutrient solution (pH 5.5) containing 10 mg (50 ppm) of 36Cl labelled 3,5,6-trichloro-2-pyridinol showed about 0.007 ppm of radioactive compounds (expressed as the pyridinol) in the plant tops after 72 h exposure. At or above pH 7, the pyridinol is readily converted to a salt in which form water solubility (4.4 g/100 ml at 25°C for the Na salt) as well as rate of absorption by the plant are enhanced relative to the free pyridinol. Regardless, soil and nutrient culture tests have again shown only negligible amounts of the compound to enter the plant and these undergo metabolism with the liberation of chloride and the formation of trivial amounts of several unidentified water soluble decomposition products (Smith et al., 1967c). Only through the use of specialized techniques has it been possible to introduce sufficient chlorpyrifos into plants to determine the extent to which the compound might be metabolized. Using the string technique, 14C chlorpyrifos was introduced into the stems of cranberry bean plants and the plants harvested at various time intervals. Subsequent analysis of the plant tissues revealed the presence principally of chlorpyrifos and its primary hydrolysis products. Under these artificial conditions, chlorpyrifos once introduced inside the stem, was distributed throughout the plant (Smith et al., 1967c). METHODS OF RESIDUE ANALYSIS Chlorpyrifos, its oxygen analogue and hydrolytic pyridinol Based on previously mentioned metabolite determinations, the main residues from chlorpyrifos treatments searched for in various tissues are chlorpyrifos, its oxygen analogue and the hydrolysed pyridinol. Since the oxygen analogue is infrequently encountered in animal or plant tissues, the principal analyses of residues are for chlorpyrifos and 3,5,6-trichloro-2-pyridinol. The latter appears in small amounts, mostly in non-fatty tissues. Analytical methods are available to determine each of these two residues individually. However, for regulatory purposes, chlorpyrifos is the most important residue to be determined. Claborn et al. (1968a,b) and Ivey and Claborn (1968) described the basic extraction and cleanup procedures using an electron capture gas chromatography method for detection of chlorpyrifos and its oxygen analogue in milk and body tissues of cattle. Claborn et al. (1970b) used a gas chromatography method and a flame photometric detector to determine chlorpyrifos and its oxygen analogue in body tissues of turkeys. Since the phosphate analogue is not searched for routinely, a quicker method for determination of chlorpyrifos only is useful. In order to do this a modification of the flame photometric gas chromatography method of Claborn et al. (1970a,b) using a hexane extraction for fat tissues was used instead of a hexane-dichloromethane extraction. The method is sensitive to 0.01 ppm. Further minor variations in the analytical method are related to extraction and cleanup procedures, depending on the nature of the substrate. Three analytical procedures are generally suitable for the various substrates when the substrates are defined as (A) water; (B) wet tissues and soil; and (C) dry or fatty tissues. The procedural scheme for methods A, B and C is outlined in Figure 2. More detailed procedures are specifically outlined as follows: Only those asterisked methods accounted for the oxygen analogue. Procedure for substrate (A), water (Rice and Dishburger, 1968; Wetters and Dishburger, 1971c). Procedure for substrate (B), wet media, such as sweet corn (Wetters, 1972c), corn forage (Wetters and Dishburger, 1971b), soil (Wetters, 1971a*,b), vegetables (Braun, 1971), liver, kidney and muscle of pig (Wetters, 1972a; Bowman and Beroza, 1967b), chicken muscle, liver, kidney and egg (Wetters, 1972b; Bowman and Beroza, 1967b), peach (Wetters and Dishburger, 1971a*), milk (Craig, 1971), cattle tissues except fat (Claborn et al., 1968a), turkey tissues except fat (Claborn et al., 1970b). Procedure for substrate (C), dry or fatty media, such as corn grain and stover (Wetters and Dishburger, 1971b; Wetters, 1972c), pig fat (Wetters, 1972a), chicken fat (Wetters, 1972b; Bowman and Beroza, 1967a), cream (Graig, 1971), cattle fat (Claborn et al., 1968a,b; Bowman and Beroza, 1967a), turkey fat (Claborn et al., 1970b). Many other references on chlorpyrifos in various media have been published (see References).
Residues of 3,5,6-trichloro-2-pyridinol may be determined by the method of McKellar and Dishburger (1970) in bovine tissues; McKellar (1971b) in corn grain, forage and stover; McKellar (1971c) in grass; Dishburger et al. (1972) in chicken tissues and eggs and McKellar (1971a) in milk and cream, at a detection level of 0.05 ppm. The general cleanup procedures shown in Figure 2 followed by a gas chromatographic determination utilizing one of the phosphorus specific detectors would be suitable for most regulatory uses. Examples of national tolerances as reported to meeting Animals Australia has a 2 ppm residue tolerance for chlorpyrifos in beef fat when cattle are treated for ticks by dip or spray. Plants Canada has a tentative 0.1 ppm residue tolerance for head lettuce, onions and carrots with 28, 60 and 60 days, respectively, required between treatment and harvest. APPRAISAL Chlorpyrifos is a non-systemic organophosphorus insecticide used to control ectoparasites of cattle, sheep and poultry; as a soil insecticide for vegetables, cereals and tobacco; and as a foliar insecticide for deciduous fruit, cereals, fodder crops, rape, cotton, some vegetables, tobacco and rice. Animal residues are found predominantly in the fat and are excreted rapidly after cessation of treatment. When cattle were given multiple dippings at the recommended dosage rates of 0.025%, the maximum total residue in fat one week after the last dipping was 1.6 ppm. When milk cows were dipped once in 0.025% chlorpyrifos, the residue in butterfat fell to 0.01 ppm in nine days and was not detectable (<0.005 ppm) after 15 days. When turkeys were confined over soil treated with chlorpyrifos (4 or 8 lb/acre) for chigger control, maximum residues of 0.16 and 0.07 ppm were found in skin and fat, respectively, one week post-treatment; the oxygen analogue was not detected in any tissues. Small amounts of the pyridinol were found in the skin, livers and kidneys of two birds with a maximum residue of 0.17 ppm in kidney after one week exposure to the second application. Extensive supervised experiments with foliar treatments on plants have shown chlorpyrifos to have little persistence and virtually no uptake occurs from either soil or foliar application. Up to 80% of applied radioactive chlorpyrifos was lost by vapourization from single leaf surfaces within the first 48 hours following treatment. There was no evidence of residue buildup on plants from repeated applications. The residues most likely to be found in either plants or animals are chlorpyrifos and its hydrolysis product 3,5,6-trichloro-2-pyridinol; the oxygen analogue or other possible metabolites are very rarely found. Both specific and general gas chromatographic methods of analysis are available which are suitable for regulatory purposes. The generalized cleanup schemes shown in Figure 2 followed by gas chromatographic determination using a phosphorus specific detector are recommended. Separate analytical procedures for determining 3,5,6-trichloro-2-pyridinol in animal tissues (liver, kidney, poultry skin), fruits and vegetables have been developed. The pyridinol is excreted rapidly in the urine and faeces of animals and there would appear to be no need to include this compound in regulatory analysis. In field experiments with plants the pyridinol is rarely found and when present constitutes only a fraction of the total residue. Although chlorpyrifos is recommended in some countries for use on rape, cereal grains and certain vegetables, there were no data available on these commodities from supervised experiments and no recommendations for tolerances could be made for these commodities. RECOMMENDATIONS TOLERANCES ppm Fat of meat of cattle 2 Chinese cabbage, grapes, kale, apples 1 Carrots, pears, tomatoes 0.5 Fat of meat of sheep, poultry, beans, eggplants, peppers, raspberries 0.2 Lettuce, rice (in husk), sugarbeets 0.1 Celery, cottonseed, cottonseed oil (crude), mushrooms, onions 0.05 Cauliflower, milk (fat basis), potatoes, red cabbage 0.01* * at or about the limit of determination FURTHER WORK OR INFORMATION REQUIRED (before tolerances can be recommended) Residue data from supervised trials for rape seed, cereal grains and any vegetables not listed. DESIRABLE 1. Elucidation of possible increased sensitivity to plasma cholinesterase depression after withdrawal from an initial dose regime. 2. Further information on residues in milk and milk products arising from dipping of dairy cattle. REFERENCES Aharonson, N., Grünberg, R. and Resnick, C. (1969) Dursban residue level in tomatoes and cotton. Ministry of Agriculture, Jaffa, Israel. Beatty, S.C. and McCollister, D.D. (1964) Results of 90-day dietary feeding studies of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate in rats. Report The Dow Chemical Co., revision. (unpublished) Blackmore, R.H. (1968a) Oral administration-dogs-Dursban final report. Hazleton Lab. Inc. (unpublished) Blackmore, R.H. (1968b) Short-term (subacute) dietary administration-rats-organophosphate insecticide Dursban - final report. Hazleton Lab. Inc. (unpublished) Bowman, M.C. and Beroza, M. (1967a) Determination of Dursban and its oxygen analogue in corn and grass by gas chromatography with flame photometric detection. J. Agr. Fd. Chem., 15(4): 651-653. Bowman, M.C. and Beroza, M. (1967b) Temperature-programmed gas chromatography of 20 phosphorus-containing insecticides on 4 different columns and its application to the analysis of milk and corn silage. J. Ass. off. analyt. Chem., 50(6): 1228-1236. Branson, D.R. and Litchfield, N.H. (1971a) Absorption, excretion and distribution of 3,5,6-trichloro-2,6-14C-2-pyridinol in rats. Report NBA-10 Dow Chemical Company. (unpublished) Branson, D.R. and Litchfield, N.H. (1971b) Absorption, excretion and distribution of O,O-diethyl O-3,5,6-trichloro-2,6-14C-2-pyridyl phosphorothioate (14C Dowco 179) in rats. Report NBA-9 Dow Chemical Company. (unpublished) Branson, D.R. and Wass, M.N. (1970) Comparative metabolism of insecticides. I. Preliminary studies of ring labelled O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate breakdown with rat liver microsomes. Report R-582 Dow Chemical Company. (unpublished) Braun, H.E. (1971) Dursban residues in vegetables. Ontario Department of Food and Agriculture, Guelph, Ontario. Personal communications to T. Haagsma. Brust, R.A., Miyazaki, S. and Hodgson, G.C. (1971) Effect of Dursban in the drinking water of chicks. J. Econ. Entomol., 64(5): 1179-1183. Burrows, I.E., Fraser, W.D., Joyce, C.A., Jenkins, G. and Eden, A. (1971a) Determination of decay curves for Dursban and its metabolite in stored tomatoes. 4211/71/369. Huntingdon Research Centre. Burrows, I.E. and Mullin, L.W. (1971b) Determination of residues of Dursban metabolite in tomatoes, grapes and apples. 4109/71/267. Huntingdon Research Centre. Burrows, I.E. and Mullin, L.W. (1971c) Determination of residues of Dursban in lettuce. 4140/71/298. Huntingdon Research Centre. Burrows, I.E., Mullin, L.W. and Eden, A. (1971d) Determination of residues of Dursban and its metabolites in raspberries. 4371/71/527. Huntingdon Research Centre. Burrows, I.E., Orme, J.P.R. and Ashton, J. (1971e) Determination of residues of Dursban and its metabolite in tomatoes. 4496/71/652. Huntingdon Research Centre. Claborn, H.V., Hoffman, R.A., Mann, H.D. and Oehler, D.D. (1968a) Residues of Dursban and its oxygen analogue in the body tissues of treated cattle. J. Econ. Entomol., 61(4): 983-986. Claborn, H.V., Mann, H.D. and Oehler, D.D. (1968b) Dursban(R) determination in milk and body tissues of cattle. J. Ass. off. analyt. Chem., 51(6): 1243-1245. Claborn, H.V., Ivey, M.C., Mann, H.D. and Oehler, D.D. (1970a) Report of residue analysis - Omental fat, muscle, kidney and liver. Report PCK-70-1. USDA-ARS-Entomol. Res. Div. Claborn, H.V., Kunz, S.E. and Mann, H.D. (1970b) Residues of Dursban in the body tissues of turkeys confined in pens containing treated soil. J. Econ. Entomol., 63(2): 422-424. Copeland, J.R. (1964) Results of 93-day dietary feeding studies of 0,0-diethyl 0-3,5,6-trichloro-2-pyridyl phosphorothioate in Beagle hounds. Biochem. Res. Lab. Report T35.12-44793-3 Dow Chemical Co. (unpublished) Coulston, F., Goldberg, L., Abraham, R., Benitz, K.F., Griffin, T.B. and Norvell, M. (1971) Final report on safety evaluation and metabolic studies on Dowco 179 (IN 151). Inst. Exp. Pathol. Toxicol., Albany Medical College. (unpublished) Coulston, F., Goldberg, L. and Griffin, T. (1972) Safety evaluation of Dowco 179 in humans. Inst. Exp. Pathol. Toxicol., Albany Medical College. (unpublished) Craig, L.F. (1971) Determination of residues of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate in milk and cream by gas chromatography. Method ACR 71.1. Dow Chemical Co. Czech, F.P. (1967) Analysis of Dursban in livestock dips and sprays. J. Ass. off. analyt. Chem., 50(4): 861-868. Dishburger, H.J. and Rice, J.R. (1965) Residues of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate in animal fat when applied as a spray on cattle. Report TA-301 Dow Chemical Co. (unpublished) Dishburger, H.J., Rice, J.R. and Muniza, R.A. (1966a) The effect of formulation on Dursban residues in omental fat of cattle following single spray application. Report TA-336 Dow Chemical Co. (unpublished) Dishburger, H.J., Rice, J.R. and Muniza, R.A. (1966b) Dursban residues in the omental fat of cattle following dip applications. Report TA-349 Dow Chemical Co. (unpublished) Dishburger, H.J., Rice, J.R. and Muniza, R.A. (1967) Dursban residues in the omental fat of cattle following two dip applications. Report TA-372 Dow Chemical Co. (unpublished) Dishburger, H.J., Pennington, J. and Rice, J.R. (1969a) Determination of O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in tobacco by gas chromatography. Report TA-430 Dow Chemical Co. (unpublished) Dishburger, H.J., Rice, J.R., McGregor, W.S. and Pennington, J. (1969b) Residues of Dursban(R) insecticide in tissues from turkeys confined on soil treated for chigger control. J. Econ. Entomol., 62(1): 181-183. Dishburger, H.J., Rice, J.R and Pennington, J. (1969c) Determination of residues of Dursban insecticide and its oxygen analogue in soil by gas chromatography. Report TA-439 Dow Chemical Co. (unpublished) Dow Chemical Co. (1964) Potentiation studies with Dursban in combination with Ruelene and malathion in rats. Report Dow Chemical Co. (unpublished) Dow Chemical Co. (1966a) Report submitted by Dow Chemical Co. (unpublished) Dow Chemical Co. (1966b) Report submitted by Dow Chemical Co. (unpublished) Dow Chemical Co. (U.K.) Ltd. (1967) Data sheets containing summarized data supporting notification of Dursban(R) insecticide for commercial use in the control of sheep ectoparasites. Submitted to the Technical Secretary, Scientific Sub-Committee, Veterinary Products Safety Precautions Scheme. Düsch, M.E., Westlake, W.E. and Gunther, F.A. (1970) Determination of Dursban insecticide in water, mud, vegetation, fish, ducks, insects and crustacea. J. Agr. Fd. Chem., 18(1): 178-179. Emerson, J.L. and Gerbig, C.G. (1970) 91 day-toxicology study in Beagle dogs treated with 3,5,6-trichloro-2-pyridinol. Report HH-263 Dow Chemical Co. (unpublished) Foschi, S. (1971) Determination del Dursban. Istituto di Patologia Vegetale, Bologna, Italy. Personal communication to K.N. Komblas. Gaines, T.B. (1969) Acute toxicity of pesticides. Toxicol. Appl. Pharmacol., 14: 515-534. Gerbig, C.G and Emerson, J.L. (1970a) Oral median lethal dose (LD50) determination of 3,5,6-trichloro-2-pyridinol in the rat. Report HH-239 Dow Chemical Co. (unpublished) Gerbig, C.G and Emerson, J.L. (1970b) Oral median lethal dose (LD50) determination of 3,5,6-trichloro-2-pyridinol in mice. Report HH-240 Dow Chemical Co. (unpublished) Graf, W.C. (1970a) Analyses of Dursban residues on cotton seeds. Dow Chemical Europe S.A. Personal communication to K. Komblas. Graf, W.C. (1970b) Dursban residue analyses on apples from SIAPA, Italy. Report DC 53. Dow Chemical Europe S.A. Graf, W.C. (1970c) Dursban residue analyses on pears from SIAPA, Italy. Report DC 54 Dow Chemical Europe S.A. Graf, W.C. and Engeler, V. (1970a) Dursban residue analyses on apples from Israel. Report DEC 55 Dow Chemical Europe S.A. Graf, W.C. and Engeler, V. (1970b) Dursban residue on tomatoes from Turkey. Report DEC 56 Dow Chemical Europe S.A. Graf, W.C. and Engeler, V. (1971a) Dowco 179 residue analysis in potatoes from France. Report DEH 2 Dow Chemical Europe S.A. Graf, W.C. and Engeler, V. (1971b) Dowco 179 residue analysis on tomatoes from Israel. Dow Chemical Europe S.A. Graf. W.C. and Engeler, V. (1971c) Dowco 179 residue analysis in apples from Israel. Report DEH 36 Dow Chemical Europe S.A. Gutenmann, W.H. St. John, Jr., L.E. and Lisk, D.L. (1968) Metabolic studies with O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate (Dursban) insecticide in a lactating cow. J. Agr. Fd. Chem., 16(1): 45-47. Hall, R.A. (1969) Dursban residues. Department of Agriculture, Australia. Personal communication to J. Tollett. Hamaguchi, H. (1970) Dowco 179 - residue analysis. Translation of analytical data furnished by Japan Plant Protection Association. Nissan Chemical Laboratory and Japan Analytical Chemical Research Laboratory. Personal communication to R.H. Ferguson. Harris, C.R. (1971) Dursban residue in onions. Canada Department of Agriculture. Personal Communication to T. Haagsma. Hollick, C.B. and Collison, R.J. (1972) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate (Dowco-179) in eggplant, tomatoes, beans and peppers treated with Dursban 4 insecticide in Turkey. Report Dow Chemical Company Ltd. (unpublished) Hunt, L.M., Gilbert, B.N. and Schlinke, J.C. (1969) Rapid gas chromatographic method for analysis of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate (Dursban) in turkey and chicken tissues. J. Agr. Fd. Chem., 17(6): 1166-1167. Hurlbert, S.H., Mulla, M.S., Keith, J.O., Westlake, W.E. and Düsch, M.E. (1970) Biological effects and persistence of Dursban(R) in freshwater ponds. J. Econ. Entomol., 63(1): 43-52. Ivey, M.C. and Claborn, H.V. (1968) Dursban oxygen analogue: Determination in fortified milk and body tissues of cattle. J. Assoc. Offic. Anal. Chem., 51(6): 1245-1246. Johnson, J.C. Jr., Bowman, M.C. and Leuck, D.B. (1969) Responses from cows fed silages containing Dursban residues. J. Dairy Sci., 52(8): 1253-1258. Kenaga, E.E. (1967) Toxicity and repellency of Dursban to the white-footed mouse (Peromyscus maniculatus). Report Dow Chemical Co. (unpublished). Leuck, D.B., Bowman, M.C. and Beck, E.W. (1968) Dursban(R) insecticide persistence in grass and corn forage. J. Econ. Entomol., 61(3): 689-690. Mann, H.D. (1966) Report of Dursban(R) insecticide residue analysis in treated cattle. Report PCK-66-2. USDA-ARS-Entomol. Res. Div. Mann, H.D. and Ivey, M.C. (1971) Report of residues analysis - Fat, skin, muscle, liver and kidney tissues of turkeys. Report PCK-71-1. USDA-ARS-Entomol. Res. Div. McCollister, S.B., Kociba, R.J., Gehring, P.J. and Humiston, C.G. (1971a) Results of two-year dietary feeding studies on Dowco(R) 179 in Beagle dogs. Report T35.12-44793-18 Dow Chemical Co. (unpublished) McCollister, S.B., Kociba, R.J., Gehring, P.J. and Humiston, C.G. (1971b) Results of two-year dietary feeding studies on Dowco(R) 179 in rats. Report, NBT35.12-44793-21 Dow Chemical Co. (unpublished) McCray, C.W. (1969) Field trials using Dursban. Letter report Animal Res. Inst., Queensland. McEwen, F.L. (1970) Tests with Dursban on onions, carrots and lettuce. University of Guelph. Personal communication to T. Haagsma. McEwen, F.L. and Frank, R. (1970) Tests with Dursban on onions, carrots and lettuce. University of Guelph. Personal communication to T. Haagsma. McKellar, R.L. (1971a) Determination of residues of 3,5,6-trichloro-pyridinol in milk and cream by gas chromatography. Method ACR 71.2. Dow Chemical Co. McKellar, R.L. (1971b) Determination of residues of 3,5,6-trichloro-pyridinol in corn grain, forage and stover by gas chromatography. Method ACR 71.19. Dow Chemical Co. McKellar, R.L. (1971c) Determination of residues of 3,5,6-trichloro-pyridinol in grass by gas chromatography. Method ACR 71.5. Dow Chemical Co. McKellar, R.L. and Dishburger, H.J. (1970) Determination of residues of 3,5,6-trichloro-2-pyridinol in bovine tissues by gas chromatography. Method ACR 70.19. Dow Chemical Co. Molello, J.A. and Sharp, L.D. (1968) Three week study on cataractogenicity of 3,5,6-trichloro-2-pyridinol as part of the dietary intake of Pekin ducklings. Report HH-134 Dow Chemical Co. (unpublished) Myers, C.M., Lewallen, L.L. and Nobe, B. (1968) Residue studies of fenthion (Baytex(R)) and Dursban(R) in central California pastures. California Vector Views, 15(5): 51-54. Norris, J.M. (1970) Potentiation Study on Dowco(R) 179 and Vapona(R) insecticide. Report Dow Chemical Co. (unpublished) Olthof, P.D.A. (1971a) Residuen van Dursban in champignons. Report R 3563. Centraal Instituut voor Voedingsonderzoek, The Netherlands. Olthof, P.D.A. (1971b) Residues of Dursban in carrots. Report R 3633. English translation from report by Centraal Instituut voor Voedingsonderzoek, The Netherlands. Rice, J.R. and Dishburger, H.J. (1968) Determination of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate in water and silt by gas chromatography. J. Agr. Fd. Chem., 16(5): 867-869. Scheuerer, W. and Hahn, I. (1972) Dursban residues on vegetables. Badische Anilin and Soda-Fabrik AG, West Germany. Communication to Dow Chemical (Nederland) N.V. Schlinke, J.C. (1970) Chronic toxicity of Dursban in chickens, 1969. J. Econ. Entomol., 63(1): 319. Schlinke, J.C., Palmer, J.S. and Hunt, L. (1969) Preliminary toxicological study of a phosphorothioate compound in turkeys. Am. J. Vet. Res., 30(9): 1705-1709. Sherman, M., Herrick, R.B., Ross, E. and Chang, M.T.Y. (1967) Further studies on the acute and subacute toxicity of insecticides to chicks. Tox. Appl. Pharmacol., 11: 49-67. SIAPA. (1971) Residue determinations on lettuce after treatment with Dursban insecticide. Centro Esperienze e Ricerche, Italy. Smith, G.N. (1966) Basic studies of Dursban insecticide. Down to Earth, 22(2): 3-7. Smith, G.N. (1968) Ultraviolet light decomposition studies with Dursban and 3,5,6-trichloro-2-pyridinol. J. Econ. Entomol., 61(3): 793-799. Smith, G.N. and Taylor, Y.S. (1970) An analytical method for the determination of 3,5,6-trichloro-2-pyridinol in animal tissues and the metabolism of the pyridinol in rats. Report OL 3132 Dow Chemical (unpublished) Smith, G.N., Watson, B.S. and Fischer, F.S. (1966) The metabolism of (14C) O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate (Dursban) in fish. J. Econ. Entomol., 59(6): 1464-1475. Smith, G.N., Watson, B.S. and Fischer, F.S. (1967a) Investigations on Dursban insecticide. Uptake and translocation of (36Cl) O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate by beans and corn. J. Agr. Fd. Chem., 15(1): 127-131. Smith, G.N., Watson, B.S. and Fischer, F.S. (1967b) Investigations on Dursban insecticide. Metabolism of (36Cl) O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate in rats. J. Agr. Fd. Chem., 15(1): 132-138. Smith, G.N., Watson, B.S. and Fischer, F.S. (1967c) Investigations on Dursban insecticide. Metabolism of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate and 3,5,6-trichloro-2-pyridinol in plants. J. Agr. Fd. Chem., 15(5): 870-877. Smith, G.N., Taylor, Y.S. and Watson, B.S. (1970) An analytical method for the determination of 3,5,6-trichloro-2-pyridinol in animal tissues and the metabolism of the pyridinol in rats. Report OL-3132 Dow Chemical Co. (unpublished) Snelson, J.T. (1969a) Dursban residues. Dept. of Primary Ind., Australia. Personal communication. Snelson, J.T. (1969b) Dursban residues. Dept. of Primary Ind., Australia. Personal communication. Stevenson, G.T. (1965) A gamebird toxicology study - acute dietary feeding of Dursban to wild type mallard ducklings. Report GH-A-122 Dow Chemical Co. (unpublished) Stevenson, G.T. (1966) A neurotoxicity study of Dursban in laying hens. Report GH-A-195 Dow Chemical Co. (unpublished) Stevenson, G.T. and Daering, K.L. (1969) The effects of Dursban insecticide (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate) on blood plasma cholinesterase in chickens. Report GH-A-410 Dow Chemical Co. (unpublished) Taylor, M.L. and Olson, K.J. (1963) Toxicological properties of O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate. Report T35.12-44793-4 Dow Chemical Co. (unpublished) Thompson, D.J., Gerbig, C.G. and Warner, S.D. (1971) Three generation reproduction and teratology study in the rat following prolonged dietary exposure to Dursban O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate. Report HH-382 Dow Chemical Co. (unpublished) Tucker, K.E.B. (1971) Dursban/Dowco 214 residues in rice and straw. Report 160 Dow Chemical (Australia) Ltd. Tucker, R.K. and Crabtree, D.G. (1970) Handbook of toxicity of pesticides to wildlife. U.S. Dept. of the Interior, Fish and Wildlife Service Resource Publication #84, p. 56-57. Tucker, K.E.B. and Zielinski, W. (1972a) Chlorpyrifos and Dowco 214 residues in Chinese cabbage. Report 175 Dow Chemical (Australia) Ltd. Tucker, K.E.B. and Zielinski, W. (1972b) Chlorpyrifos and Dowco 214 residues in Chinese kale. Report 174 Dow Chemical (Australia) Ltd. Tucker, K.E.B. and Zielinski, W. (1972c) Residues of chlorpyrifos in Chinese kale. Report 179 Dow Chemical (Australia) Ltd. Tucker, K.E.B. and Zielinski, W. (1972d) Residues of chlorpyrifos and Dowco 214 in tobacco. Report 180 Dow Chemical (Australia) Ltd. USAEHA. (1968) Report U.S. Army Environmental Hygiene Agency. Watts, R.M. (1968) Residues of chlorpyrifos in cattle. Dept. of Agriculture, Australia. Communication to M.K. Astill. Wetters, J.H. (1971a) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate and O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphate in soils by gas chromatography with flame photometric detection. Method ACR 71.3. Dow Chemical Co. Wetters, J.H. (1971b) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in soils by gas chromatography using flame photometric detection and direct injection. Method ACR 71.20. Dow Chemical Co. Wetters, J.H. (1972a) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in pig tissues by gas chromatography with flame photometric detection. Method 72.1. Dow Chemical Co. Wetters, J.H. (1972b) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in chicken tissues and eggs by gas chromatography with flame photometric detection. Method 72.3. Dow Chemical Co. Wetters, J.H. (1972c) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in sweet corn by gas chromatography with flame photometric detection. Method ACR 72.9. Dow Chemical Co. Wetters, J.H. and Dishburger, H.J. (1971a) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate and O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphate in peaches by gas chromatography with flame photometric detection. Method ACR 71.14. Dow Chemical Co. Wetters, J.H. and Dishburger, H.J. (1971b) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in corn by gas chromatography with flame photometric detection. Method ACR 71.18. Dow Chemical Co. Wetters, J.H. and Dishburger, H.J. (1971c) Determination of residues of O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate in water by gas chromatography with flame photometric detection. Method ACR 71.21. Dow Chemical Co. Winterlin. W., Moilanen, K. and Burgoyne, W.E. (1968) Residues of Dursban insecticide following mosquito control applications. Down to Earth, 24(2): 34-37. Wit, S.L. (1970) Residues of Dursban in cabbage. Report 49/70 Tox RoB. English translation from report by State Institute for the Public Health, The Netherlands. Wit, S.L. (1971) Residues of O,O-diethyl O-(3,5,6-trichloro-pyridine-(2)-yl) phosphorothioate (Dursban) for the control of cutworms in lettuce under glass. Report 63/71 Tox-RoB. English translation from report by Rijks Instituut voor de Volksgezondheid, The Netherlands.
See Also: Toxicological Abbreviations Chlorpyrifos (ICSC) Chlorpyrifos (WHO Pesticide Residues Series 5) Chlorpyrifos (Pesticide residues in food: 1977 evaluations) Chlorpyrifos (Pesticide residues in food: 1981 evaluations) Chlorpyrifos (Pesticide residues in food: 1982 evaluations) Chlorpyrifos (Pesticide residues in food: 1983 evaluations) Chlorpyrifos (JMPR Evaluations 1999 Part II Toxicological)