Glycine

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with restrictions (only two dose levels, limited documentation (clinical signs, gross pathology))

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

carcinogenicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

only two dose levels, limited documentation (clinical signs, gross pathology)

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

The carcinogenicity of glycine was assessed in rats by application in the drinking water.

GLP compliance:

no

Species:

rat

Strain:

Fischer 344/DuCrj

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Co. Japan, Inc. Atsugi
- Age at study initiation: 6 wks
- Housing: in plastic cages (3/sex/cage)
- Diet (ad libitum): pellet diet (CRF-1, Charles River, Japan)
- Water (ad libitum)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 2
- Humidity (%): 55 ± 5

Route of administration:

oral: drinking water

Vehicle:

unchanged (no vehicle)

Duration of treatment / exposure:

108 wks

Frequency of treatment:

continuously via drinking water

Dose / conc.:

2.5 other: % (nominal in water)

Remarks:

equivalent to 1534 mg/kg bw/day for males and 1587 mg/kg bw/day for females (actual ingested)

Dose / conc.:

5 other: % (nominal in water)

Remarks:

equivalent to 3280 mg/kg bw/day for males and 3082 mg/kg bw/day for females (actual ingested)

No. of animals per sex per dose:

50

Control animals:

other: drinking water without test substance

Details on study design:

- Dose selection rationale: on the basis of a preliminary subacute experiment (not further specified)

Observations and examinations performed and frequency:

BODY WEIGHT: Yes
- Time schedule for examinations: every one or two weeks

FOOD CONSUMPTION: Yes
- Time schedule for examinations: every four weeks

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at week 108
- How many animals: all survivors
- Parameters checked: white/red blood cells, heamoglobin (Hb), haematocrit (Ht)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at week 108
- How many animals: all survivors
- Parameters checked: glutamic oxalacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine phosphokinase (CPK), blood urea nitrogen (BUN), creatinine (CRE), Urea (UA), total protein (TP), albumin/globulin rate (A/G rate), blood glucose (BG), phospholipid (PL), total cholesterol (Tch), triglyceride (TG), natrium, potassium, calcium, chloride

URINALYSIS: Yes
- Parameters checked: leukocytes, nitrite, urobilinogen, protein, pH, ketone, glucose

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (all animals, including those that died or were sacrificed upon becoming moribund during the experiment)
HISTOPATHOLOGY: Yes (all organs were examined and samples were fixed in 10% buffered neutral formalin, embedded in paraffin, and routinely stained with hematoxylin and eosin; histological analysis of tumors in renal pelvis)

Statistics:

Wilcoxon t-test or chi-squared test

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

5% dose group: slightly increased mortality in females compared to controls

Mortality:

mortality observed, treatment-related

Description (incidence):

5% dose group: slightly increased mortality in females compared to controls

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group: slight dose-dependent inhibition of mean body weight gain in both sexes

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not examined

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

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

5% dose group: significant changes of Hb and HT in males and females

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group, males and females: significantly changes in CPK, BUN, TP, Tch and TG were observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group: papillary necrosis and transitional hyperplasia was observed in male and female animals (no cases seen in the controls)

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group, females: kidney papillomas were seen, which were not observed in controls

Details on results:

CLINICAL SIGNS AND MORTALITY
Mortality of females of the high dose group was slightly increased when compared to controls (number of surviving females and males: 31, 46, 41 and 40, 45, 40 for the high, low, control group).

BODY WEIGHT AND WEIGHT GAIN
2.5 and 5% dose group, males and females: Mean body-weights were slightly but dose-dependently inhibited when compared to controls over the hole study period (only growth curves were given).

HAEMATOLOGY
5% dose group, males: A slight but not significant increase in white blood cells was observed. The Hb was significantly increased when compared to controls.
5% dose group, females: A slight but not significant increase in white blood cells and a slight but not significant decrease in red blood cells was observed. The Hb and HT were significantly decreased when compared to controls.
2.5% dose group, females: A slight but not significant increase in white blood cells and a slight but not significant decrease in red blood cells was observed.

CLINICAL CHEMISTRY
2.5 and 5% dose group, males and females: The CPK values were significantly decreased when compared to controls in both sexes and both dose groups. BUN was significantly increased in both sexes of the 5% dose group.
2.5 and 5% dose group, females: A significant and dose-dependent decrease of TP and Tch and a significant and dose-dependent increase in TG was observed.
Some sporadic effects on isolated parameters were observed which were considered as not treatement-related (see Table 1 "Any other information on results incl. tables").

URINALYSIS
No significant differences in urinalysis data between treated and control male and female animals were observed. The only change was increased numbers of erythrocytes in glycine-treated male and female rats. This was expected from haemorrhage in necrotic renal papillae (see histopathology).

ORGAN WEIGHTS
No significant effects on the relative organ weights were observed in the treated groups when compared to controls.

HISTOPATHOLOGY: NON-NEOPLASTIC
The incidences of papillary necrosis in the kidney of male rats fed 2.5 and 5% glycine were 4 and 7%, while the corresponding figures for females were 30 and 32%. Since necrosis of the renal papillae was limited to glycine-treated rats it was very likely linked to the induction of papillomas. Transitional hyperplasia in renal pelvis was observed in 2 female animals of the 2.5% dose group. Calcification indices in kidney were 39, 32 and 42% of the 0, 2.5 and 5% groups. Endometrial polyps in the uterus were increased in both treatment group females (see Table 2/3 "Any other information on results incl. tables").
Overall, for the males, tumors were found in 90% of the high dose group, 97% of the low dose group and in 90% of the controls, while the corresponding figures for females were 59, 65 and 39%. These tumors were considered to be spontaneous in nature (see Table 3 "Any other information on results incl. tables").

HISTOPATHOLOGY: NEOPLASTIC (if applicable)
One renal cell tumor was seen in one male of the high dose group. Renal pelvis papillomas were found in 8% of the low dose group and in 6% of the high dose group females, but not in the controls. Since spontaneous tumors in this tissues are very rare, they were considered as treatment-related (see Table 3 "Any other information on results incl. tables").

Dose descriptor:

LOAEL

Remarks:

carcinogenicity

Effect level:

1 561 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: histopathology

Dose descriptor:

NOAEL

Remarks:

carcinogenicity

Effect level:

< 1 561 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: histopathology

Critical effects observed:

not specified

Table 1: Clinical chemistry data for rats treated with the test substance (mean values ± standard deviation)

	Males			Females
Group (%)	0	2.5	5	0	2.5	5
No. of rats	40	45	40	41	46	31
GOT (K-U)	122 ± 72	120 ± 57	114 ± 27	130 ± 53	117 ± 40	136 ± 68
GPT (K-U)	51 ± 34	55 ± 37	53 ± 12	59 ± 22	56 ± 12	66 ± 16
LDH (K-U)	2879 ± 772	2710 ± 972	2514 ± 819	2572 ± 1369	2666 ± 836	2556 ± 952
ALP (KA-U)	16 ± 6	15 ± 5	17 ± 5	19 ± 9	19 ± 5	25 ± 7*
CPK (IU/L)	754 ± 321	563 ± 217*	509 ± 197*	706 ± 392	545 ± 160*	446 ± 158*
BUN (mg/dL)	18 ± 3	17 ± 2	21 ± 3*	18 ± 2	17 ± 2	21 ± 3*
CRE (mg/dL)	0.6 ± 0.07	0.5 ± 0.10*	0.5 ± 0.11*	0.5 ± 0.06	0.5 ± 0.06	0.5 ± 0.07
UA (mg/dL)	2.3 ± 1.5	1.9 ± 0.6	2.5 ± 1.3	2.8 ± 1.8	2.0 ± 0.9*	2.5 ± 1.7
TP (g/dL)	6.9 ± 0.5	6.6 ± 0.3*	6.8 ± 0.4	7.5 ± 0.7	7.1 ± 0.5*	6.8 ± 0.7*
A/G (rate)	0.6 ± 0.09	0.7 ± 0.06*	0.7 ± 0.06*	0.8 ± 0.07	0.8 ± 0.06	0.8 ± 0.09
BG (mg/dL)	131 ± 46	127 ± 30	140 ± 51	136 ± 35	127 ± 15	125± 27
PL (mg/dL)	216 ± 62	205 ± 54	214 ± 61	211 ± 37	205 ± 36	202 ± 34
Tch (mg/dL)	150 ± 51	130 ± 39	129 ± 42	117 ± 22	104 ± 19*	98 ± 17*
TG (mg/dL)	175 ± 82	162 ± 59	198 ± 78	154 ± 73	209 ± 102*	226 ± 105*
Na (mEq/L)	144 ± 2	141 ± 2*	143 ± 2	141 ± 3	139 ± 1*	141 ± 4
K (mEq/L)	5.3 ± 0.7	4.6 ± 0.8*	5.3 ± 1.0	5.0 ± 0.9	4.4 ± 0.5*	4.7 ± 0.8
Ca (mg/dL)	11.1 ± 0.9	10.3 ± 0.5*	11.2 ± 1.1	11.2 ± 1.4	10.6 ± 0.6*	10.7 ± 1.1
Cl (mEq/L)	99 ± 1.9	101 ± 2.3*	99 ± 3.5	99 ± 2.0	98 ± 2.3	97± 4.3

*p<0.05, compared to control

Table 2: Histological findings for the urinary system for rats treated with the test substance

		No. of affected rats (%)
		Males			Females
Test substance concentration (%)		0	2.5	5	0	2.5	5
Effective No. of rats		40	45	40	41	46	31
Urinary system
Kidney	renal cell tumor	0	0	1(2)	0	0	0
	papillary necrosis	0	2(4)	3(7)	0	14(30)	10(32)
	calcification	0	0	0	16(39)	15(32)	13(42)
Pelvis	transitional hyperplasia	0	0	0	0	2(4)	0
	transitional cell tumor	1(2)	0	0	0	0	0
	papilloma	0	0	0	0	4(8)	2(6)

Table 3: Organ distribution and histological diagnosis for tumors in rats treated with the test substance

		No. of affected rats (%)
		Males			Females
Test substance concentration (%)		0	2.5	5	0	2.5	5
Effective No. of rats		40	45	40	41	46	31
Tumor development		36(90)	44(97)	36(90)	27(59)	30(65)	12(39)
Genital system
Testis	interstitial cell tumor	35(87)	44(97)	36(90)	-	-	-
Prostate	adenoma	1(2)	2(4)	2(4)	-	-	-
	calcification	-	-	-	-	-	-
Uterus	endometrial polyp	-	-	-	5(12)	18(39)	12(38)
	endometrial sarcoma	-	-	-	0	1(2)	0
	adenocarcinoma	-	-	-	1(2)	0	0
Mammary gland	firboma	0	0	0	1(2)	0	0
	adenoma	0	0	0	0	1(2)	1(3)
Endocrine system
Pituitary gland	adenoma	9(22)	6(13)	7(17)	13(31)	12(26)	9(29)
Thyroid gland	C-cell adenoma	4(10)	1(2)	3(7)	4(9)	3(6)	2(6)
	C-cell carcinoma	0	2(4)	1(2)	1(2)	1(2)	0
	papillary adenoma	1(2)	0	1(2)	0	2(4)	0
	papillary adenocarcinoma	0	1(2)	0	0	0	0
Adrenal gland	pheochromocytoma	5(13)	7(15)	5(13)	0	1(2)	0
Pancreas	islet cell tumor	6(15)	1(2)	2(4)	0	1(2)	0
Digestive system
Liver	neoplastic nodule	1(2)	0	0	0	0	1(3)
Respiratory system
Lung	adenoma	0	2(4)	0	0	1(2)	0
Urinary system
Kidney	renal cell tumor	0	0	1(2)	0	0	0
	transitional cell tumor	1(2)	0	0	0	0	0
	papilloma	0	0	0	0	4(8)	2(6)
Urinary bladder	papilloma	0	0	0	0	0	0
Body cavity
Peritoneum	mesothelioma	2(5)	2(4)	1(2)	0	0	0
Hematopoietic system
Leukemia		4(10)	6(13)	3(7)	6(14)	9(17)	3(9)
Integument, musculoskeletal system
Subcutis	fibroma	3(8)	0	0	4(10)	2(4)	0
	lipoma	0	1(2)	0	0	0	1(2)
	preputial/clitoral gland tumor	3(6)	2(4)	2(4)	0	0	1(2)
	fibrosarcoma	0	0	1(2)	0	0	0

Data source

Reference

Reference Type:

publication

Title:

Carcinogenicity study of glycine in Fischer 344 rats

Author:

Kitahori, Y. et al.

Year:

1994

Bibliographic source:

J Toxicol Pathol 7: 471-480

Materials and methods

Principles of method if other than guideline:

The carcinogenicity of glycine was assessed in rats by application in the drinking water.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Glycine

EC Number:

200-272-2

EC Name:

Glycine

Cas Number:

56-40-6

Molecular formula:

C2H5NO2

IUPAC Name:

glycine

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

Fischer 344/DuCrj

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Co. Japan, Inc. Atsugi
- Age at study initiation: 6 wks
- Housing: in plastic cages (3/sex/cage)
- Diet (ad libitum): pellet diet (CRF-1, Charles River, Japan)
- Water (ad libitum)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 2
- Humidity (%): 55 ± 5

Administration / exposure

Route of administration:

oral: drinking water

Vehicle:

unchanged (no vehicle)

Duration of treatment / exposure:

108 wks

Frequency of treatment:

continuously via drinking water

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2.5 other: %

Remarks:

nominal in water

Doses / concentrations 2

Dose / conc.:

5 other: %

Remarks:

nominal in water

Doses / concentrations 3

Dose / conc.:

1 534 mg/kg bw/day (actual dose received)

Remarks:

males

Doses / concentrations 4

Dose / conc.:

3 280 mg/kg bw/day (actual dose received)

Remarks:

males

Doses / concentrations 5

Dose / conc.:

1 587 mg/kg bw/day (actual dose received)

Remarks:

females

Doses / concentrations 6

Dose / conc.:

3 082 mg/kg bw/day (actual dose received)

Remarks:

females

No. of animals per sex per dose:

50

Control animals:

other: drinking water without test substance

Details on study design:

- Dose selection rationale: on the basis of a preliminary subacute experiment (not further specified)

Examinations

Observations and examinations performed and frequency:

BODY WEIGHT: Yes
- Time schedule for examinations: every one or two weeks

FOOD CONSUMPTION: Yes
- Time schedule for examinations: every four weeks

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at week 108
- How many animals: all survivors
- Parameters checked: white/red blood cells, heamoglobin (Hb), haematocrit (Ht)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at week 108
- How many animals: all survivors
- Parameters checked: glutamic oxalacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine phosphokinase (CPK), blood urea nitrogen (BUN), creatinine (CRE), Urea (UA), total protein (TP), albumin/globulin rate (A/G rate), blood glucose (BG), phospholipid (PL), total cholesterol (Tch), triglyceride (TG), natrium, potassium, calcium, chloride

URINALYSIS: Yes
- Parameters checked: leukocytes, nitrite, urobilinogen, protein, pH, ketone, glucose

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (all animals, including those that died or were sacrificed upon becoming moribund during the experiment)
HISTOPATHOLOGY: Yes (all organs were examined and samples were fixed in 10% buffered neutral formalin, embedded in paraffin, and routinely stained with hematoxylin and eosin; histological analysis of tumors in renal pelvis)

Statistics:

Wilcoxon t-test or chi-squared test

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

5% dose group: slightly increased mortality in females compared to controls

Mortality:

mortality observed, treatment-related

Description (incidence):

5% dose group: slightly increased mortality in females compared to controls

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group: slight dose-dependent inhibition of mean body weight gain in both sexes

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not examined

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

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

5% dose group: significant changes of Hb and HT in males and females

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group, males and females: significantly changes in CPK, BUN, TP, Tch and TG were observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group: papillary necrosis and transitional hyperplasia was observed in male and female animals (no cases seen in the controls)

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

2.5 and 5% dose group, females: kidney papillomas were seen, which were not observed in controls

Details on results:

CLINICAL SIGNS AND MORTALITY
Mortality of females of the high dose group was slightly increased when compared to controls (number of surviving females and males: 31, 46, 41 and 40, 45, 40 for the high, low, control group).

BODY WEIGHT AND WEIGHT GAIN
2.5 and 5% dose group, males and females: Mean body-weights were slightly but dose-dependently inhibited when compared to controls over the hole study period (only growth curves were given).

HAEMATOLOGY
5% dose group, males: A slight but not significant increase in white blood cells was observed. The Hb was significantly increased when compared to controls.
5% dose group, females: A slight but not significant increase in white blood cells and a slight but not significant decrease in red blood cells was observed. The Hb and HT were significantly decreased when compared to controls.
2.5% dose group, females: A slight but not significant increase in white blood cells and a slight but not significant decrease in red blood cells was observed.

CLINICAL CHEMISTRY
2.5 and 5% dose group, males and females: The CPK values were significantly decreased when compared to controls in both sexes and both dose groups. BUN was significantly increased in both sexes of the 5% dose group.
2.5 and 5% dose group, females: A significant and dose-dependent decrease of TP and Tch and a significant and dose-dependent increase in TG was observed.
Some sporadic effects on isolated parameters were observed which were considered as not treatement-related (see Table 1 "Any other information on results incl. tables").

URINALYSIS
No significant differences in urinalysis data between treated and control male and female animals were observed.

ORGAN WEIGHTS
No significant effects on the relative organ weights were observed in the treated groups when compared to controls.

HISTOPATHOLOGY: NON-NEOPLASTIC
The incidences of papillary necrosis in male rats fed 2.5 and 5% glycine were 4 and 7%, while the corresponding figures for females were 30 and 32%. Since necrosis of the renal papillae was limited to glycine-treated rats it was very likely linked to the induction of papillomas. Transitional hyperplasia in renal pelvis was observed in 2 female animals of the 2.5% dose group. Calcification indices in kidney were 39, 32 and 42% of the 0, 2.5 and 5% groups (see Table 2 "Any other information on results incl. tables").

HISTOPATHOLOGY: NEOPLASTIC (if applicable)
For the males, tumors were found in 90% of the high dose group, 97% of the low dose group and in 90% of the controls, while the corresponding figures for females were 59, 65 and 39%. These tumors were considered to be spontaneous in nature (see Table 3 "Any other information on results incl. tables").
One renal cell tumor was seen in one male of the high dose group. Kidney papillomas were found in 8% of the low dose group and in 6% of the high dose group females, but not in the controls. Since spontaneous tumors in this tissues are very rare, they were considered as treatment-related (see Table 3 "Any other information on results incl. tables").

Effect levels

1

Dose descriptor:

LOAEL

Effect level:

1 561 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: clinical chemistry; histopathology

2

Dose descriptor:

NOAEL

Effect level:

< 1 561 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

histopathology: neoplastic

histopathology: non-neoplastic

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1: Clinical chemistry data for rats treated with the test substance (mean values± standard deviation)

	Males			Females
Group (%)	0	2.5	5	0	2.5	5
No. of rats	40	45	40	41	46	31
GOT (K-U)	122 ± 72	120 ± 57	114 ± 27	130 ± 53	117 ± 40	136 ± 68
GPT (K-U)	51 ± 34	55 ± 37	53 ± 12	59 ± 22	56 ± 12	66 ± 16
LDH (K-U)	2879 ± 772	2710 ± 972	2514 ± 819	2572 ± 1369	2666 ± 836	2556 ± 952
ALP (KA-U)	16 ± 6	15 ± 5	17 ± 5	19 ± 9	19 ± 5	25 ± 7*
CPK (IU/L)	754 ± 321	563 ± 217*	509 ± 197*	706 ± 392	545 ± 160*	446 ± 158*
BUN (mg/dL)	18 ± 3	17 ± 2	21 ± 3*	18 ± 2	17 ± 2	21 ± 3*
CRE (mg/dL)	0.6 ± 0.07	0.5 ± 0.10*	0.5 ± 0.11*	0.5 ± 0.06	0.5 ± 0.06	0.5 ± 0.07
UA (mg/dL)	2.3 ± 1.5	1.9 ± 0.6	2.5 ± 1.3	2.8 ± 1.8	2.0 ± 0.9*	2.5 ± 1.7
TP (g/dL)	6.9 ± 0.5	6.6 ± 0.3*	6.8 ± 0.4	7.5 ± 0.7	7.1 ± 0.5*	6.8 ± 0.7*
A/G (rate)	0.6 ± 0.09	0.7 ± 0.06*	0.7 ± 0.06*	0.8 ± 0.07	0.8 ± 0.06	0.8 ± 0.09
BG (mg/dL)	131 ± 46	127 ± 30	140 ± 51	136 ± 35	127 ± 15	125± 27
PL (mg/dL)	216 ± 62	205 ± 54	214 ± 61	211 ± 37	205 ± 36	202 ± 34
Tch (mg/dL)	150 ± 51	130 ± 39	129 ± 42	117 ± 22	104 ± 19*	98 ± 17*
TG (mg/dL)	175 ± 82	162 ± 59	198 ± 78	154 ± 73	209 ± 102*	226 ± 105*
Na (mEq/L)	144 ± 2	141 ± 2*	143 ± 2	141 ± 3	139 ± 1*	141 ± 4
K (mEq/L)	5.3 ± 0.7	4.6 ± 0.8*	5.3 ± 1.0	5.0 ± 0.9	4.4 ± 0.5*	4.7 ± 0.8
Ca (mg/dL)	11.1 ± 0.9	10.3 ± 0.5*	11.2 ± 1.1	11.2 ± 1.4	10.6 ± 0.6*	10.7 ± 1.1
Cl (mEq/L)	99 ± 1.9	101 ± 2.3*	99 ± 3.5	99 ± 2.0	98 ± 2.3	97± 4.3

*p<0.05, compared to control

Table 2: Histological findings for the urinary system for rats treated with the test substance

		No. of affected rats (%)
		Males			Females
Test substance concentration (%)		0	2.5	5	0	2.5	5
Effective No. of rats		40	45	40	41	46	31
Urinary system
Kidney	renal cell tumor	0	0	1(2)	0	0	0
	papillary necrosis	0	2(4)	3(7)	0	14(30)	10(32)
	calcification	0	0	0	16(39)	15(32)	13(42)
Pelvis	transitional hyperplasia	0	0	0	0	2(4)	0
	transitional cell tumor	1(2)	0	0	0	0	0
	papilloma	0	0	0	0	4(8)	2(6)

Table 3: Organ distribution and histological diagnosis for tumors in rats treated with the test substance

		No. of affected rats (%)
		Males			Females
Test substance concentration (%)		0	2.5	5	0	2.5	5
Effective No. of rats		40	45	40	41	46	31
Tumor development
Genital system
Testis	interstitial cell tumor	36(90)	44(97)	36(90)	27(59)	30(65)	12(39)
Prostate	adenoma	35(87)	44(97)	36(90)	-	-	-
	calcification	1(2)	2(4)	2(4)	-	-	-
Uterus	endometrial polyp	-	-	-	5(12)	18(39)	12(38)
	endometrial sarcoma	-	-	-	0	1(2)	0
	adenocarcinoma	-	-	-	1(2)	0	0
Mammary gland	firboma	0	0	0	1(2)	0	0
	adenoma	0	0	0	0	1(2)	1(3)
Endocrine system
Pituitary gland	adenoma	9(22)	6(13)	7(17)	13(31)	12(26)	9(29)
Thyroid gland	C-cell adenoma	4(10)	1(2)	3(7)	4(9)	3(6)	2(6)
	C-cell carcinoma	0	2(4)	1(2)	1(2)	1(2)	0
	papillary adenoma	1(2)	0	1(2)	0	2(4)	0
	papillary adenocarcinoma	0	1(2)	0	0	0	0
Adrenal gland	pheochromocytoma	5(13)	7(15)	5(13)	0	1(2)	0
Pancreas	islet cell tumor	6(15)	1(2)	2(4)	0	1(2)	0
Digestive system
Liver	neoplastic nodule	1(2)	0	0	0	0	1(3)
Respiratory system
Lung	adenoma	0	2(4)	0	0	1(2)	0
Urinary system
Kidney	renal cell tumor	0	0	1(2)	0	0	0
	transitional cell tumor	1(2)	0	0	0	0	0
	papilloma	0	0	0	0	4(8)	2(6)
Urinary bladder	papilloma	0	0	0	0	0	0
Body cavity
Peritoneum	mesothelioma	2(5)	2(4)	1(2)	0	0	0
Hematopoietic system
Leukemia		4(10)	6(13)	3(7)	6(14)	9(17)	3(9)
Integument, musculoskeletal system
Subcutis	fibroma	3(8)	0	0	4(10)	2(4)	0
	lipoma	0	1(2)	0	0	0	1(2)
	preputial/clitoral gland tumor	3(6)	2(4)	2(4)	0	0	1(2)
	fibrsarcoma	0	0	1(2)	0	0	0