Phenol, isopropylated, phosphate (3:1)

Repeated dose toxicity: inhalation

Administrative data

Endpoint:

sub-chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

No information

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non-GLP, not conducted according to recognised guideline, but fully detailed method and results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The experimental regimen for MIL.H.19457C, MIL-H-19457B, and MIL-H-22072B was identical. Male and female Fischer.344 rats, male Golden Syrian hamsters, and male and female New Zealand White rabbits were exposed to hydraulic fluid aerosol on a continuous basis for 90 days in Thomas Dome inhalation chambers. In addition, sham exposed control groups of animals of each corresponding speies and sex were maintained in an inhalation exposure chamber.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Phenol, isopropylated, phosphate (3:1)

EC Number:

273-066-3

EC Name:

Phenol, isopropylated, phosphate (3:1)

Cas Number:

68937-41-7

Molecular formula:

CXHYO4P X and Y are variable dependant on the molecular component.

IUPAC Name:

Phenol, isopropylated, phosphate (3:1)

Details on test material:

- Name of test material (as cited in study report): Durad MP280 (MIL-H-19457B)
- Substance type: Phenol, isopropylated, phosphate (3- 1) 100%
- Physical state: Liquid
- Lot/batch No.: 67-5

No other data specified within the report.

Specific details on test material used for the study:

Referenced as MIL-H-19457B

Test animals

Species:

other: rats, hamsters and rabbits.

Strain:

other: Fischer-344 rats, Golden Syrian hamsters and New Zealand White rabbits.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: No data.
- Age at study initiation: Fischer 344 rats (8-10 weeks of age); Golden hamsters (8-10 weeks of age); rabbits not specified.
- Weight at study initiation: No data.
- Fasting period before study: No data.
- Housing: Stainless steel cages with wire mesh floors.
- Diet (e.g. ad libitum): ad libitum but the rats were fasted during the 18 h period preceding their scheduled sacrifice at the end of study.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data.
- Humidity (%): No data.
- Air changes (per hr): No data.
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark.

IN-LIFE DATES: From: 7 July 1982 To: 3 October 1982

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

not specified

Remarks on MMAD:

MMAD / GSD: No data.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Thomas Dome inhalation chamber.
- Method of holding animals in test chamber: No data.
- Source and rate of air: no data.
- Method of conditioning air: no data.
- System of generating particulates/aerosols: Concentrations of MIL-H-194S7e and Mll-H-194578 were monitored using a TSI Aerosol Photometer, Model 5150 (TSllnc., Minneapolis, MN). Automatic timers were use to acquire three samples of four-minutes each from each chamber for photometric analysis. Chamber supply air analyses were used to establish baseline values during the remaining periods of time.
- Temperature, humidity, pressure in air chamber: no data.
- Air flow rate: no data.
- Air change rate: no data.
- Method of particle size determination: Via a cascade impactor (Partcle Fractonating Sampler, Anersn Samplers, Inc., Atlanta, GA), samples were taken daily from each chamber for particle size analysis. Aluminum stage collecor plates were weighed before and after sampling and the mass of each calculated. The particle size mass median diameter and stndard geometric deviation were calculated from thes data.

TEST ATMOSPHERE
- Brief description of analytical method used: Analysis routinely performed during the course of the 90-day study using a Liquid chromotography (HPLC) technique.
- Samples taken from breathing zone: Yes

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentrations of MlL-H-19457B were monitored using a TSI Aerosol Photometer, Model 5150 (TSllnc., Minneapolis, MN). Automatic timers were use to acquire three samples of four-minutes each from each chamber for photometric analysis. Chamber supply air analyses were used to establish baseline values during the remaining periods of time.

Duration of treatment / exposure:

90 days.

Frequency of treatment:

Continuous inhalation.

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
10mg/m3
Basis:
nominal conc.

Doses / concentrations 2

Remarks:

Doses / Concentrations:
100mg/m3
Basis:
nominal conc.

No. of animals per sex per dose:

Each chamber (representative of a dose level) housed 20 male and 20 female Fischer-344 rats, 20 male Golden Syrian hamsters and 4 male and 4 female New Zealand rabbits.

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: The Naval Medical Resach Institute/Toxicology Detachment (NMRI/TD) provided information from industrial hygiene surveys aboard ship which indicated aerosol concentrations of 25 mg/ml maximum (8 mg/ml average). These surveys were conducted for the glycol-based materials; however, it is reasonable to assume that this concentration of aerosol is similar to concentrations produced when triarylphosphate fluids are use in similar shipboard operations. Base on these data, a continuous expore to 10 mg/m3 was chosen as the low concentration expoure and 100 mg/ml as the high concentration exposure. The low concentration simulated shipboard conditions while the high concentration might be expected to produce a toxic effect.
- Rationale for animal assignment (if not random): No data.
- Rationale for selecting satellite groups: No data.
- Post-exposure recovery period in satellite groups: No data.
- Section schedule rationale (if not random): No data.

Positive control:

No data.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data
- Time schedule: Hourly for signs of toxicity.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data.

BODY WEIGHT: No data
- Time schedule for examinations: Weekly.

FOOD CONSUMPTION: No data

FOOD EFFICIENCY: No data

WATER CONSUMPTION: No data

HAEMATOLOGY: Yes - only for rats.
- Time schedule for collection of blood: Blood samples were taken from ten male and ten female rats per exposure group for clinical chemistry determinations at 42 days. At the end of the 90-day exposure hematologic and clinical chemistry determinations were made using blood samples that were taken at necropsy from male and female rats that were not sampled at 42 days exposure.
- Animals fasted: No data
- Anaesthetic used for blood collection: No data
- How many animals: 20 - 10 male and 10 female rats per exposure group.
- Parameters checked below were examined:
Haematocrit; Haemoglobin; RBC; WBC; WBC Differential; Mean Corpuscular Volume (MCV); Mean Corpuscular Hemoglobin (MeH); Mean Corpuscular Hemoglobin Concentration (MCHC).

CLINICAL CHEMISTRY: Yes only for rats.
- Time schedule for collection of blood: Blood samples were taken from ten male and ten female rats per exposure group for clinical chemistry determinations at 42 days. At the end of the 90-day exposure hematologic and clinical chemistry determinations were made using blood samples that were taken at necropsy from male and female rats that were not sampled at 42 days exposure.
- Animals fasted: No data
- How many animals: 20 - 10 male and 10 female rats per exposure group.
- Parameters checked below were examined:
Calcium; Albumin/Globulin; Total Protein; Alkaline Phosphatase; SGOT; Creatinine; BUN.

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: Yes CNS Examination
- Time schedule for examinations: After 2 weeks, 4 weeks, 8 weeks and at test termination.
- Dose groups that were examined: Five male and five female rats from the high concentration and the control group.
- Battery of functions tested: sensory and motor activity:

Autonomic Signs; As each rat was taken from its home cage, it was evaluated for the presence of diarrhea, lacrimation (including chromodacryorrhea), nasal discharge, salivation, exophthalmus, and urinary incontinence. Presence of any of these were recorded; and absence of all autonomic
signs was noted with a zero (Irwin, 1968).

Tail Tip Curl Reflex: Holding the rat in the right hand under the forelimbs, the tail was stroked toward the tail tip with the left forefinger beginning at a point about one third of its length from the tip, and hesitating at a point about 1 to 1 1/2 inches from the tip. In normal rats, the tail tip curled,
more or less strongly, around the finger. The strength of the response was rated on a scale of 0 (no response), 1 (present but not stronglrisk), or 2 (strongly curled, brisk response). If a rat began to struggle, it was returned to its cage, then retested. A maximum of three attempts were allowed.

Hind Foot Position/Foot Drop: Normal rats supported under the forelimbs typically show one of two hind limb positions: (a) the feet drawn up against the body, the ankles flexed, the legs parallel, and the toes extended in a relatively relaxed fashion; or (b) the legs splayed outward, theankles extended in a relaxed manner or spread. Drooping feet, crosse legs, and/or curled toes are abnormal (Snyder and Braun, 1977). Hind limb poition was scored and recorded as 0 (sign absent) or 1 (sign present).

Lateral Hop: Each rat was held around the thorax in one hand and the rat's tail taken in the other. One hind foot was placed on the edge of a bench top while the tail was pulled downward slightly to put weight on the foot. When the rat was tilted in the direction of the foot in contact with the bench top, it normally would hop on that one foot in that direcion. The test was done both to the left and right and the response was scored 0 (no hop or depressed response) or 1 (presnt/normalresponse) (Marshall and Teitelbaum, 1974) and recorded.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table in attached background material: 90 day continuous inhalation tox results tables.pdf )
HISTOPATHOLOGY: Yes (see table in attached background material: 90 day continuous inhalation tox results tables.pdf )

Other examinations:

None detailed.

Statistics:

Data Analysis: The design for body weight analysis was repeated measures with a blocking effect of sex (male, female). If there was an overall significant F (p < 0.05), Ryan-Einot-Gabriel-Welsch Range Test (SAS Institute, Inc., 1985) was used to find pairwise comparisons. The organ weights and bloo data were statistically analyzed via a two factorial analysis of variance. The factors were exposure concentration and sex. If overall F was significant (p<0.05) pairwise comparisons were done using the Ryan-Einot-Gabriel-Welsch Range Test. An Armitage test of linear trend (SAS Institute, Inc., 1985) and a Chi-square test for proportions (Dixon, 1985) were used to analyze the pathology data. The same group of control animals, according to species and sex, were used for comparison with groups of animals exposed to hydraulic fluids, appropriate corrections wereincorporated into the computerized statistical procedures.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Mortality:

mortality observed, treatment-related

Description (incidence):

See Details on results

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

See Details on results

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY: Natural deaths among hamsters and rats were few and sporadic. Several deaths occurred, particularly in the high concentration MIL-H-19457B female rat group during an interim bleeding regimen. The bleeding technique was altered slightly and additional deaths were avoided. Signs of toxic stress were noted in the high concentration MIL-H-19457B rabbit group as early as three days after exposure initiation. The rabbits became anorexic and lethargic. This was followed by cachexia accompanied by head droop prior to death. Paralysis was not noted. All rabbits from this group died by the 49th expoure day. Mortality also occurred in several of the other rabbit groups, including controls. Pasteurella multocida infection was the cause in some of these deaths, including one rabbit that died from the low concentration MIL-H-19457B group.
Kyphosis was noted in rats early in the 100 mg/m3 MIL-H-19457B exposure. This persisted throughout the 90 days. These groups of rats also had rough coats and a general unkempt appearance.

BODY WEIGHT AND WEIGHT GAIN: Male rats exposed to MIL-H-19457B showed no exposure related differences in growth. Female rats exposed to 100 mg/m3 exhibited an erratic growth curve that was generally lower than and statistically different (p<0.01) from the controls. Hamsters exposed to 100 mg/m3 also showed slightly depressed weight gain particularly during the latter stages of the exposure when weight loss occurred, however, the mean weights were never significantly different from those of the control group. Rabbits were rapidly losing weight by the second week of exposure to 100 mg/ml and were all dead by the eighth week. The weight of rabbits exposed to 10 mg/ml were unaffected throughout the exposure.

FOOD CONSUMPTION: No data.

FOOD EFFICIENCY: No data.

WATER CONSUMPTION: No data.

OPHTHALMOSCOPIC EXAMINATION: No data.

HAEMATOLOGY: Statistically significant group means were noted for a few parameters, however, marked differences did not occur.

CLINICAL CHEMISTRY: The group mean values were statistically significantly different for several parameters, yet the actual magnitude of most of those differences appeared to be relatively small. Some of the more marked group differences occurred in calcium, SGPT, and alkaline phosphatase values.

URINALYSIS: No data.

NEUROBEHAVIOUR: MIL-H-19457B didn't substantially alter the responses measured by the hindfoot drop test or the lateral hop test. A general decline in performance in the tail tip curl test was noted in male rats exposed to MIL-H-19457B. Results of this test in female rats were erratic. Statistically significant differences in tail tip curl test responses could only be demonstrated when data for both sexes were combined. The loss of test subjects toward the termination of the exposure hindered a conclusive interpretation.

ORGAN WEIGHTS: In a number of the groups the heart, spleen. and brain weights of rats exposed to the three hydraulic fluids were statistically different (p≤0.01) from control values. No remarkable trends were noted in any of these values. Significant elevations in liver weight occurred in male rats exposed to MIL-H-19457B. The relative liver weight of male rats exposed to 100 mg/m3 was increased more than 40% for MIL-H-19457B, compared with the control group. Exposure to 10mg/m3 of the registered substance produced a less dramatic increase: 4% - MlL-H-19457B). No difference between the relative liver weights of the control group and rats exposed to 10 mg/m3 MIL-H-19457B was noted. Groups of rats having significantly elevated relative liver weights also had increase absolute liver weights. Analyses of female rat organ weights indicated elevated liver weight in animals exposed to MIL-H-19457B. The increase relative liver weights observed were comparable between male and female rats. An increase of approximately 10% occurred in the livers of females exposed to 10 mg/m3 of the registered subtance, while an increase of 45% occurred in the livers of females exposed to MIL-H-19457B at the high concentrations.

GROSS PATHOLOGY: Gross lesions of note in male rats at termination of exposure were testicular atrophy in 25% of the 100 mg MIL-H-19457B/m3 group, adrenal enlargement in 33% of the 100 mg/m3 MlL-H-19457B. Grossly enlarged adrenal glands were also observed in 25% and 53% respectively of the 10 and 100 mg/m3 MIL-H-19457B groups of female rats. The gross examination of rabbits and hamsters at the conclusion of the exposure period disclosed no remarkable lesions.

HISTOPATHOLOGY: RATS: The control male and female rats (20 per sex) incidences of pulmonary interstitial inflammation were respectively, 35 % and 40%. The pituitary gland of 19% of the control males (3/16) displayed small populations of enlarged adenohypophyseal cells with abundant, eosnophilic cytoplasm. Six male and two female control rats had adrenocortical fatty change. Mild pancreatic atrophy was obsrved in 2/20 (10%) of the control females.
- Nose: Mild goblet cell hyperplasia of the nasal mucos was noted in 6/20 (30%) of the males, and 1/20 (5%) of the females in the high concentration group. No low concentration subject and only 1/20 (5%) of the control males exhibited this change.
- Lungs: Perivascular cuffing and interstitial inflammation were inflammatory changes which were considered components of a continuum of pulmonary interstitial effec. Where both perivascular cuffing and interstital inflammation were present in the same animal, only one diagnosis (inflammation) was use for incidence data. When considered together, and in both sexes, these mild to moderate lung changes were obsrved in 18/40 (45%)control, 20140 (50%) lC, and 15/40 (38%) HC rats. Following combination of all pulmonary inflammatory diagnoses, no expoure relationship was evident.
- Heart: Minimally severe foci of myocardial inflammation and degeneration were presnt in 9/19 (47%) LC male rats and in 1/20 (5%) HC rats. Similar myocardiallesions were not seen in control males or females.
- Liver: Mild hepatocellular swelling was observed in 3/20 (15%) males and 4/20 (20%) female rats in the HC group.
- Kidney: Renal papillary necrosis (16/20) and tubular cell fatty change (12/20) were observed in the HC females only. These lesions were not documented in control female rats or male rats and were considered to be a distinct concentration-related effec of expoure to MIL-H-19457B in females.
- Adrenal: Distinct adrenocortical fatty change was observed in all males and females in both exposure groups, and generally with concentration-dependent severity, Six of twenty (30%) male controls, and 2/20 (10%) female controls also exhibited adrenocortical fatty change but only minimal severity.
- Ovary: Hypertrophy of ovarian interstitial cells was noted in 18/19 (95%) HC and 6/20 (30%) LC females.
- Testis: Moderate to severe degeneration of testicular seminiferous tubules was found in all HC males (100%). It was not present in LC or control males. Also, mild intersttial cell hyperplasia was detected in 4120 (20%) HC males.
- Pituitary: Twelve of nineteen (63%) HC males exhibited small populations of enlarged adenohypophysal cells with abundant, eonophilic cyoplasm.

HISTOPATHOLOGY: RABBITS:
Morphologic diagnoses and incidences of histopathologic changes in both sexes of control rabbits and rabbits exposed to 10 mg/m3 and 100 mg/ml concentrations of MIL-H-19457B. The primary histopathologic changes observed in rabbits used as control were chronic nasal inflammation in three males, and lymphocytic interstitial inflammation in the lungs of two males. A lesion which deserves comment was centrilobular to panlobular hepatocellular fatty change in the livers of all four female rabbits exposed to the high concentration of MIL-H-19457B. Similar fat accumulations were not observed in hepatocytes of males exposed to the high concentration of MIL-H-19457B. Although not included in Table 13, degenerative neurologic lesions were seen in the spinal cord of one control male rabbit, and one female rabbit expose to the low concentration of MIL-H-19457B.

Effect levels

Key result

Dose descriptor:

NOEC

Effect level:

10 mg/L air (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

clinical biochemistry

clinical signs

gross pathology

mortality

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

MIL-H-19457B appears to be the more toxic of the two triarylphosphate compounds based on the mortality data, gross findings and histopathology. The toxicity of each compound at 10 mg/m1, the predicted shipbord concentration, appeared to be minimal.

Table 2: Animal Mortality During Exposure To Hydraulic Fluid:

Species and Sex	Controls	MIL-H-19457B
		10mg/m3	100mg/m3
Rats, Male (20)a	5.0	0.0	0.0
Rats, Female (20)	10.0	0.0	25.0
Hamsters, Male (20)	0.0	5.0	0.0
Rabbits, Male (4)	0.0	0.0	100.0b
Rabbits, Female (4)	25.0	25.0b	100.0

a = Original number of animals/group in parentheses.

b = Includes a death from Pasteurella multocida infection.

Table 3: Mean Scores of CNS Examination of Rats Exposed for 90 Days To Hydraulic Fluids (a):

Male Rats
Tail Tip Curl	2 Weeks	4 Weeks	8 Weeks	Termination
Control	1.9 ± 0.1	1.6 ± 0.2	1.8 ± 0.1	1.8 ±0.1
MIL-H-19457B	1.7 ± 0.1	1.9 ± 0.1	1.2 ± 0.2	1.0±0.2
Hind Foot Drop
Control	0	0	0	0
MIL-H-19457B	0	0	0	0
Lateral Hop
Control	1.0 ± 0.0	1.0 ± 0.0	1.0 ± 0.0	1.0 ± 0.0
MIL-H-1945B	1.0 ± 0.0	1.0 ± 0.0	0.6 ± 0.2	0.9 ± 0.1
Female Rats
Tail Tip Curl	2 Weeks	4 Weeks	8 Weeks	Termination
Control	1.7 ± 0.2	1.0 ± 0.0	1.5 ± 0.3b	1.5 ± 0.3b
MIL-H-19457B	1.5 ± 0.5	2.0 ± 0.0	1.0 ±0.0c	1.5 ± 0.5d
Hind Foot Drop
Control	0	0	0b	0b
MIL-H-19457B	0	0	0c	0d
Lateral Hop
Control	1.0 ± 0.0	1.0 ± 0.0	1.0 ± 0.0b	1.0 ± 0.0b
MIL-H-19457B	1.0 ± 0.0	1.0 ± 0.0	0.5 ± 0.5c	1.0 ± 0.0d

a: Mean±S.E.M.; n = 5 unless described otherwise; units described in test.

b: n = 4

c: n = 3

d: n = 2

Table 4: Haematology Values (a) of Rats After Exposure to MIL-H-19457B for 90 Days:

MALE RATS	Control	MIL-H-19457B
	(N = 8)	10mg/m3 (N=10)	100mg/m3 (N=10)
RBC (10^6 cells/µL)	8.22 ± 0.04	8.05 ± 0.10	8.18 ± 0.11
WBC (10^3 cells/µL)	5.19 ± 0.34	5.48 ± 0.41	7.36 ± 0.43b,c
HCT (%)	41.90 ± 0.42	40.10 ± 0.51	40.88 ± 0.45
HGB (g/dL)	14.64 ± 0.17	14.37 ±0.16	14.63 ± 0.23
MCV (fl)	50.96 ± 0.35	49.82 ± 0.28	49.96 ± 0.25
MCH (pg)	17.80 ± 0.16	17.86 ± 0.14	17.87 ± 0.14
MCHC (g/dL)	34.93 ± 0.13	35.86 ± 0.41	35.77 ± 0.27
FEMALE RATS	Control	MIL-H-194578B
	(N=8)	10mg/m3 (N=10)	100mg/m3 (N=5)
RBC (10^6 cells/µL)	7.72 ± 0.07	7.70 ± 0.06	7.43 ±0.13
WBC (10^3 cells/µL)	3.64 ± 0.20	3.98 ± 0.26	4.24 ±0.28
HCT (%)	41.97 ± 0.50	47.29 ± 1.86b	38.90 ± 0.38b,c
HGB (g/dL)	14.57 ± 0.15	14.09 ±0.14b	13.36 ± 0.20bc
MCV (fL)	54.35 ±0.44	53.49±0.29b	52.38 ±0.46
MCH (pg)	18.87±0.14	18.29±0.84	17.98±0.05b
MCHC	34.73±0.28	34.14±0.14	34.34±0.23

a: Mean±S.E.M

b: Mean different from control at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

c: Statistically different from 10mg/m3 group at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

Table 5. Clinical Chemistry Values (a) of Male Rats after 42 or 90 Days of Exposure to MIL-H-19457B:

42 Days	Control	MIL-H-19457B
	(N=10)	10mg/m3 (N=10)	100mg/m3 (N=10)
BUN (m/dL)	22.68±1.31	20.94±1.34	19.21±1.45b
Creatinine (mg/dL)	0.75±0.04	0.72±0.04	0.65±0.05
Total Protein (g/dL)	8.27±0.27	8.54±0.12	8.15±0.19
Albium (g/dL)	3.79±0.37	4.72±0.31	5.07±0.27b
Globulin (g/dL)	4.48±0.48	3.82 ±0.30	3.09±0.27b
Alb/Glob Ratio	1.04±0.22	1.36±0.18	1.76±0.16b
Calcium (g/dL)	8.36±0.56	14.83±1.76b	21.26±0.71b,c
SGOT/AST (IU/L)	69.78±15.27	54.15±3.55	63.82±5.62
SGPT/ALT (IU/L)d	--------------	--------------	-----------------
Alkaline Phos (IU/L)	166.40±15.27	54.15±3.55	63.82±5.62
90 Days	Control	MIL-H-19457B
	(N=10)	10mg/m3 (N=10)	100mg/m3
BUN(mg/dL)	13.87±0.71	14.49±0.49	15.86±1.04
Creatinine (mg/dL)	0.56±0.03	0.65±0.02b	0.63±0.02
Total Protein (g/dL)	9.15±0.10	9.50±0.15	9.11±0.12
Albium (g/dL)	4.85±0.06	4.68±0.05b	4.62±0.06b
Globulin (g/dL)	4.29±0.08	4.82±0.15b	4.49±0.09
Alb/Glob Ratio	1.13±0.02	0.98±0.03b	1.03±0.02b
Calcium (g/dL)	8.59±0.47	9.92±0.41	8.30±0.19c
SGOT/AST (IU/L)	58.29±3.01	58.96±4.02	59.06±2.21
SGPT/ALT (IU/L)d	25.07±1.20	23.49±1.28	45.14±2.27b,c
Alkaline Phos (IU/L)	57.00±2.03	48.00±1.26b	40.40±1.34b,c

a: Mean±S.E.M

b: Mean different from control at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

c: Statistically different from 10mg/m3 group at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

d: This test was not performed due to an insufficient volume of serum.

Table 6: Clinical Chemistry Values (a) of Female Rats after 42 or 90 Days of Exposure to MIL-H-19457B:

42 Days	Control	MIL-H-19457B
	(N=10)	10mg/m3 (N=10)	100mg/m3 (N=10)
BUN (m/dL)	26.34±1.34	25.04±0.82b	18.67±0.96b,c
Creatinine (mg/dL)	0.70±0.03	0.75±0.04	0.81±0.05
Total Protein (g/dL)	9.79±0.56	10.23±0.27	10.46±0.53
Albium (g/dL)	5.78±0.13	5.09±0.32b	5.40±0.28
Globulin (g/dL)	4.13±0.61	5.21 ±0.38	5.32±0.47
Alb/Glob Ratio	1.57±0.21	1.03±0.13	1.00±0.10b
Calcium (g/dL)	11.00±0.32	12.93±0.73b	13.13±0.54b
SGOT/AST (IU/L)	71.18±7.90	83.04±4.21	80.51±5.85
SGPT/ALT (IU/L)d	--------------	--------------	-----------------
Alkaline Phos (IU/L)	114.40±6.09	102.60±2.09	75.40±2.79b,c
90 Days	Control	MIL-H-19457B
	(N=10)	10mg/m3 (N=10)	100mg/m3 (N=9)
BUN(mg/dL)	19.36±0.90	17.13±0.99	28.79±1.85b,c
Creatinine (mg/dL)	0.61±0.03	0.62±0.02	0.59±0.02
Total Protein (g/dL)	0.62±0.26	8.53±0.16	8.24±0.22
Albium (g/dL)	5.04±0.06	5.01±0.06	4.69±0.13b,c
Globulin (g/dL)	3.58±0.21	3.53±0.12	3.55±0.17
Alb/Glob Ratio	1.44±0.07	1.43±0.04	1.35±0.08
Calcium (g/dL)	12.21±0.38	11.88±0.18	11.44±0.29
SGOT/AST (IU/L)	77.42±4.37	85.60±8.94	81.50±6.45
SGPT/ALT (IU/L)d	27.05±2.17	32.17±4.12	36.10±2.18b
Alkaline Phos (IU/L)	34.60±1.87	34.20±2.00	27.44±1.89b,c

a: Mean±S.E.M

b: Mean different from control at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

c: Statistically different from 10mg/m3 group at p<0.05, using a two factorial analysis of variance and the Ryan-Einot-Gabriel-Welsch Range Test.

d: This test was not performed due to an insufficient volume of serum.

- Further tables can be found as attached background material.

Applicant's summary and conclusion

Conclusions:

It was dertermined that MIL-H-19457B was the more toxic of the two triarylphosphate compounds based on the mortality data, gross findings and histopathology. The toxicity of each compound at 10 mg/ml, the predicted shipbord concentration, appeared to be minimal.

Executive summary:

It was determined that MIL-H-19457B was the more toxic of the two triarylphosphate compounds based on the mortality data, gross findings and histopathology. The toxicity of each compound at 10 mg/m3, the predicted shipbord concentration, appeared to be minimal.