Bis[[R-(R*,S*)]-β-hydroxy-α-methylphenethyl)methylammonium] sulphate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Information from a published industry source (NTP), restrictions in design and/or reporting but otherwise adequate for assessment

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1986

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Ten Fischer 344/N rats were given diets containing 0, 125, 250, 500, 1000, or 2000 ppm of the test substance. Animals were observed twice per day and weighed once per week. After an exposure period of 13 weeks, animals were necropsied, blood was taken for the determination of packed cell volume and pupil diameters were measured.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Bis[[R-(R*,S*)]-β-hydroxy-α-methylphenethyl)methylammonium] sulphate

EC Number:

205-154-4

EC Name:

Bis[[R-(R*,S*)]-β-hydroxy-α-methylphenethyl)methylammonium] sulphate

Cas Number:

134-72-5

Molecular formula:

C10H15NO.1/2H2O4S

IUPAC Name:

bis([(1R,2S)-1-hydroxy-1-phenylpropan-2-yl](methyl)azanium) sulfate

Details on test material:

- Name of test material (as cited): L-ephedrine sulfate
- Lot No.: 926EW
- Purity: >99%
- Storage: in the dark at 25°C

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories (Portage, MI)
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: group mean males 129 - 158 g and females 109 - 115 g
- Housing: 5 animals per cage, Polycarbonate cages (Hazleton Systems, Inc., Aberdeen, MD), Reeman Spun-bonded polyester filters (Snow Filtration, Cincinnati, OH); Aspen wood chips as bedding.
- Diet: ad libitum, NIH 07 Rat and Mouse Ration (Zeigler Bros., Garners, PA)
- Water: ad libitum, Automatic watering system (Edstrom Idnustries, Waterford, WI)
- Acclimation period: 20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 26.6
- Humidity (%): 20 - 48
- Air changes (per hr): 120
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: 1979-12-31 to 1980-04-04

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Premix mixed with feed in a twin-shell blender for 5 min with intensifier bar and then 10 min without intensifier bar
- Maximum storage time: 14 days
- Storage temperature of food: Room temperature

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
125, 250, 500, 1000, 2000 ppm
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
Males: 4.6, 9.6, 22.2, 43.4, 87.4 mg/kg bw/day (mean)
Basis:
actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:
Females: 8.1, 16.6, 35.6, 67.2, 144.9 mg/kg bw/day (mean)
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, sham-exposed

Examinations

Observations and examinations performed and frequency:

- CAGE SIDE OBSERVATIONS AND DETAILED CLINICAL OBSERVATIONS: twice daily
- FOOD CONSUMPTION AND COMPOUND INTAKE: measured once a week. Food consumption for each animal determined and divided by final body weight to estimate the daily intake of the test substance in mg/kg body weight/day
- BODY WEIGHT: measured once a week.
- OPHTHALMOSCOPIC EXAMINATION: Pupil diameter measured at the end of the study.

Sacrifice and pathology:

- Necropsy and histologic exam performed on all animals. Tissues examined: tissue masses, regional lymph node, blood smear, skin, mandibular lymph node, mammary gland, salivary gland, thigh muscle, sciatic nerve, bone marrow, costochondral junction (rib), thymus, larynx, trachea, lungs and bronchi, heart, thyroid gland, parathyroids, esophagus, stomach, duodenum, jejunum, ileum, colon, rectum, mesenteric lymph node, liver, gallbladder, pancreas, spleen, kidneys, adrenal glands, urinary bladder, seminal vesicles/prostate/tested or ovaries/uterus, nasal cavity, brain, pituitary gland, eyes, external and middle ear and spinal cord.
- Adrenal gland and heart weight measured for control and highest dose groups.

Other examinations:

Determination of packed cell volume. Blood was taken from the external jugular vein at the scheduled kill.

Statistics:

t-test was used to compare packed cell volumes and organ weights of the controls with animals exposed to the test substance.

Results and discussion

Results of examinations

Details on results:

MORTALITY AND CLINICAL SIGNS
- No mortality observed in all exposed animals.
- No mortality observed in the control group.
- Animals that received 2000 ppm were hyperexitable and had rough coats.

BODY WEIGHT AND WEIGHT GAIN
- Final mean body weights of rats that received 1,000 or 2,000 ppm were 20% and 23% lower than those of the controls for males and 10% and 17% lower for females.
- The mean final body weights for males were 359, 347, 339, 315, 286, and 277 g for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean final body weights for females were 201, 189, 184, 184, 180, and 167 g for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean body weight gains for males were 218, 189, 199, 186, 146, and 142 g for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean body weight gains for females were 90, 80, 74, 73, 68, and 52 g for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.

FOOD CONSUMPTION AND COMPOUND INTAKE
- Feed consumption by dosed and control groups was generally comparable.
- The mean feed consumption of males in week 6 was 15.1, 14.2, 14.4, 15.4, 14.1, and 12.7 g per animal per day for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean feed consumption of males in week 12 was 14.8, 12.9, 13, 14, 12.4, and 12.1 g per animal per day for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean feed consumption of females in week 6 was 9.7, 9.3, 8.6, 8.9, 8.8, and 8.5 g per animal per day for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The mean feed consumption of females in week 12 was 12.7, 12.2, 12.2, 13.1, 12.1, and 12.1 g per animal per day for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.

OPHTHALMOSCOPIC EXAMINATION
- The mean pupil diameters for dosed and control animals were comparable.
- The pupil diameter, right and left combined, of males was 1.46, 1.46, 1.29, 1.26, 1.06, and 1.49 mm for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
- The pupil diameter, right and left combined, of females was 1.04, 1.34*, 1.01, 0.86, 1.01, and 0.97 mm for the control, 125, 250, 500, 1000, and 2000 ppm exposed animals respectively.
* = statistically significant difference compared to the control group (p<0.05).

ORGAN WEIGHTS
- The mean adrenal gland weights of males were 39.9 and 37.9 mg for the control and 2000 ppm exposed animals respectively.
- The relative adrenal gland weights of males were 1.11 and 1.37 for the control and 2000 ppm exposed animals respectively, which is a statistically significant difference (p<0.01).
- The mean heart weights of males were 0.828 and 0.772 g for the control and 2000 ppm exposed animals respectively.
- The relative heart weights of males were 2.3 and 2.79 for the control and 2000 ppm exposed animals respectively.
- The mean adrenal gland weights of females were 51.3 and 37.1 mg for the control and 2000 ppm exposed animals respectively, which is a statistically significant difference (p<0.01).
- The relative adrenal gland weights of females were 2.55 and 2.27 for the control and 2000 ppm exposed animals respectively.
- The mean heart weights of females were 0.66 and 0.451 g for the control and 2000 ppm exposed animals respectively, which is a statistically significant difference (p<0.01).
- The relative heart weights of females were 3.27 and 2.69 for the control and 2000 ppm exposed animals respectively.

GROSS PATHOLOGY and HISTOPATHOLOGY
No compound related histopathological effects were observed.

OTHER FINDINGS: Packed cell volume
- The mean packed cell volume of male rats that received 2,000 ppm was 6% greater than that of the controls
- The mean packed cell volume in males was 47.1 and 49.7 % for the control and 2000 ppm exposed animals respectively, which is a statistically significant difference (p=0.004)
- The mean packed cell volume in females was 46.2 and 46.6 % for the control and 2000 ppm exposed animals respectively.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

500 other: ppm (nominal in diet)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Lower body weights of 10% and 17% for males and 20% and 23% for females compared to the control, at 1000 and 2000 ppm, and the clinical signs observed at 2000 ppm.

2

Dose descriptor:

NOAEL

Effect level:

22 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Clinical signs and lower final body weights in comparison with controls.

3

Dose descriptor:

NOAEL

Effect level:

35 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Clinical signs and lower final body weights in comparison with controls

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In a 90-day oral repeated dose toxicity study in Fischer 344/N rat exposed via diet, the NOAELs were determined to be 500 ppm (nominal in diet); 22 mg/kg bw/day (actual ingested for males) and 35 mg/kg bw/day (actual ingested for females).

Executive summary:

Ten Fischer 344/N rats were given diets containing 0, 125, 250, 500, 1000, or 2000 ppm of the test substance. Animals were observed twice per day and weighed once per week. After an exposure period of 13 weeks, animals were necropsied, blood was taken for the determination of packed cell volume and pupil diameters were measured. None of the rats died before the end of the studies. Final mean body weights of rats that received 1000 or 2000 ppm were 20% and 23% lower than those of the controls for males and 10% and 17% lower for females. Feed consumption by dosed and control groups was generally comparable. Rats that received 2000 ppm were hyperexcitable and had rough coats. No compound-related histopathologic effects were observed. The mean pupil diameters for dosed and control animals were comparable. The mean packed cell volume of male rats that received 2000 ppm was 6% greater than that of the controls. The relative adrenal gland weight of male rats that received 2000 ppm was significantly greater than that of the controls, and the adrenal gland weight and heart weight of female rats that received 2000 ppm were significantly lower than those of the controls. Based on the a lower body weights compared to controls and clinical signs observed the NOAELs were determined to be 500 ppm (nominal in diet), 22 mg/kg bw/day (actual ingested for males) and 35 mg/kg bw/day (actual ingested for females).