SUMILIZER GP

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 Dec 2007 - 28 Jul 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Objective of study:

absorption

distribution

excretion

mass balance

metabolism

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 417 (Toxicokinetics)

Version / remarks:

adopted in 1984

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 417 (Toxicokinetics)

Version / remarks:

adopted in 2010

Deviations:

yes

Remarks:

age of the animals not reported

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss Federal Office of Public Health, Consumer protection directorate, Notification authority for chemicals, Bern, Switzerland

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

442-450-4

EC Name:

-

Cas Number:

203255-81-6

Molecular formula:

C42H61O4P

IUPAC Name:

6-(3-(3-tert-Butyl-4-hydroxy-5-methylphenyl)propoxy)-2,4,8,1 0-tetra-tert-butyldibenz(d,f)(1,3,2)dioxaphosphepin

Radiolabelling:

yes

Test animals

Species:

rat

Strain:

other: CD (SD) IGS

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Kissleg, Germany
- Weight at study initiation: 238 ± 21 g (males) and 217 ± 21 g (femlaes)
- Housing: In groups of 1-3/sex under conventional hygienic conditions in Makrolon cages with standard soft wood bedding during acclimation. Individually in metabolism cages during the course of the study.
- Diet: Pelleted 3433 Kliba rat maintenance diet (Provimi Kliba AG, Kaiseraugstm Switzerland), ad libitum
- Water: Tap water, ad libitum
- Acclimation period: at least 5 days
- Health status: The health status of the treated animals was checked visually at daily intervals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: aqueous methyl cellulose

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Stock solutions of 14C-labeled test items were prepared by mixing aliquot amounts of unlabeled test item with 14C-labeled test item (MOH or TBPH-label) in 10 mL of hexane (10 mg/kg bw, groups 3, 4, 7, and 8) or dichloromethane (1000 mg/kg bw, groups 5 and 6) in a volumetric flask.

MOH-labelled test item, 10 mg/kg bw: (groups 3+4)
9.95 mL of the stock solution were transferred into a volumetric flask and the solvent was evaporated under a nitrogen stream. The test item was pre-dissolved in 1.5 mL of DMSO and 0.5% methocel solution was added to a total volume of 30 mL under stirring. Test item concentration and homogeneity was verified by LSC measurements of triplicate samples. The resulting test item concentration in the application solution was 0.98 mg/g.

TBPH-labelled test item, 10 mg/kg bw: (groups 7+8)
9.85 mL of the stock solution were transferred into a volumetric flask and the solvent was evaporated under a nitrogen stream. The test item was pre-dissolved in 1.5 mL of DMSO and 0.5% methocel solution was added to a total volume of 30 mL under stirring. Test item concentration and homogeneity was verified by LSC measurements of triplicate samples. The resulting test item concentration in the application solution was 0.98 mg/g

TBPH-labelled test item, 1000 mg/kg bw: (groups 5+6)
The complete stock solution was transferred into a 100 mL round bottom flask and 30 mL 0.5% methocel solution were added. The dichloromethane solvent was evaporated in a nitrogen stream under vigorous stirring and further 10-12 mL methocel solution added to replace loss by evaporation and to reach a suitable consistency for administration. The concentration of the test item solution was determined by LSC to be 67.8 mg/g. The formulation was kept under permanent stirring until administration.

Duration and frequency of treatment / exposure:

single administration

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

10 mg/kg bw/day (nominal)

Remarks:

equivalent to 9.80 ± 0.18 mg/kg bw in males and 9.98 ± 0.30 mg/kg bw in females;
low dose MOH-label, groups 3 (males) and 4 (females), corresponding to 25 MBq (0.67 mCi)

Doses / concentrations 2

Dose / conc.:

10 mg/kg bw/day (nominal)

Remarks:

equivalent to 9.65 ± 0.19 mg/kg bw in males and 10.10 ± 0.30 mg/kg bw in females;low dose TBPH-label, groups 7 (males) and 8 (females), corresponding to 25 MBq (0.67 mCi)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

equivalent to 992 ± 45 mg/kg bw in males and 991 ± 73 mg/kg bw in females;
high dose TBPH-label, groups 5 (males) and 6 (females), corresponding to 50 MBq (1.35 mCi)

No. of animals per sex per dose / concentration:

4

Control animals:

no

Details on study design:

- Dose selection rationale: Comparison of the excretion balance after administration of 2 labels (MOH and TBPH) and between a high dose (1000 mg/kg bw) and a low dose (10 mg/kg bw).

Details on dosing and sampling:

TOXICOKINETIC / PHARMACOKINETIC STUDY (Absorption, distribution, excretion)
- Tissues and body fluids sampled: urine, faeces, cage washes, blood, carcass and tissues. As tissues, the adrenal glands, blood, brain, epididymes , eyes , fat, femur, heart, kidney, large intestinal tract, liver, lung , muscle , ovaries, pancreas , pituitary gland, plasma , prostate gland , sciatic nerve, skin (back region), small intestinal tract, spinal cord, spleen, stomach, testes , thymus, thyroid gland, urinary bladder and uterus were collected. Additionally, the contents of the stomach, large intestine and small intestine of each animal were pooled and analysed.
- Time and frequency of sampling:
urine: 0-8, 8-24, 24-48, 48-72, 72-96, 96-120, 120-144, and 144-168 h after administration
faeces: 0-24, 24-48, 48-72, 72-96, 96-120, 120-144, and 144-168 h after administration
blood: at sacrifice after 168 h
organs/tissues/carcass: at sacrifice after 168 h
cage wash: 0-8 and 0-24 h after administration

SAMPLE PROCESSING
CAGE WASH: At the end of the each interval 0-8 and 0-24 h collection intervals, the metabolism cages were rinsed with a small amount of purified water (about 5-10 mL) to remove any remaining droplets of urine. The rinsings were added to the initially sampled urine and the radioactivity was evaluated. At the end of the study, the cages were washed with ethanol, water/acetone (50/50, v/v) and/or appropriate solvnts for the determination of any residual radioactivity.
URINE: Subsamples of urine were mixed at room temperature with liquid scintillation counting (LSC) cocktail (IRGA-SAFE PLUSTM, Perkin Elmer) analysed by LSC. Counting was performed using a Packard TRI-CARBTM Liquid Scintillation Analyser (model 2500 TR).
FAECES: Faeces samples were homogenised wet by addition of water (about 1+2, w/v). After homogenization, the radioactivity was determined from appropriate aliquots after solubilisation with tissue solubiliser (Solvable, Perkin Elmer).
CARCASS: The carcass was homogenized and the radioactivity was determined from appropriate aliquots of the homogenate after solubilisation with tissue solubiliser (Solvable, Perkin Elmer).
ORGANS / BLOOD: Subsamples were digested with tissue solubiliser (Solvable, Perkin Elmer), followed by determination of radioactivity.
INTESTINAL TRACT: The contents of the stomach, large intestine and small intestine of each animal were pooled, homogenised wet by addition of water (about 1+2, w/v and radioactivity was determined from appropriate aliquots after solubilisation with tissue solubiliser (Solvable, Perkin Elmer).
PLASMA: Plasma samples were directly measured for radioactivity.
BONE: The femur was combusted and radioactivity was determined thereafter.

EXTRACTION OF FAECES POOLS
Faeces samples of the 24 and 48 h collection time points were pooled per group and extracted 3 times with neural acetonitrile, followed by a further extraction with 0.1 M HCl/Acetonitrile (2:8). The 0-24 h faeces pools of the group 3 animals were extracted with ethyl acetate before the acidic acetonitrile extraction. As this additional step did not improve the recovery, all other pools were processed without ethyl acetate extraction. The extracts were combined, concentrated and analysed by HPLC.

PURIFICATION OF URINE POOLS
Four pools were generated of the individual urine samples of the group 3 and 4 animals, collection intervals 0-8 and 8-24 h with 2 mL urine per animal and collection interval. A Separtis Isolute RP18 solid phase extraction column1 (5 g sorbent mass) was pre-conditioned with 0.1% aqueous ammonium acetate. 8 mL of urine per group were transferred onto the column and eluted with 0.1% aqueous ammonium acetate (approximately 11 mL per pool) and 100% methanol (4.1 - 4.4 mL). Total activity in all fractions was determined, the total recovery was 88- 102%. The methanol fractions were analysed by HPLC.

METABOLITE CHARACTERISATION STUDIES
- Tissues and body fluids sampled: urine and faeces
- Time and frequency of sampling:
urine: 0-8, 8-24 and 24-48 h after administration
faeces: 0-24 and 24-48 h after administration
- From how many animals: sampled were pooled for each sex and dose group
- Method types for identification: HPLC with UV detector and radiodetector, Liquid scintillation counting (LSC)
- Limits of detection and quantification: Limits of detection (LOD) and limits of quantification (LOQ) for liquid scintillation counting were calculated according to L.A. Currie (Anal. Chem. 40, 586 (1968)) from the specific radioactivity of the test item.

ANALYTICAL METHODS :

Measurement of radioactivity
Radioactivity in all specimens was determined on Packard liquid scintillation counters (e.g. TRICARB 2500 TR, 2550 TR/LL or 2900 TR) using the Transformed Spectral Index of the External Standard Spectrum (tSIE) method for quench correction. All measurements were performed as far as possible at least in duplicate for a counting time allowing a counting error below 1% or for a maximal counting time of 10 minutes. All measurements were performed with scintillation background correction. In case duplicates differed by more than 10% from the mean value, as far as possible re-analyses were performed using fresh aliquots (except for measurements below 100 dpm).

The following scintillation mixtures were used:
A: Irga-Safe Plus (Perkin Elmer)
B: Solvable (Perkin Elmer)
Water: Milli-Q water (Milli-Q plus 185, Millipore)

HPLC analysis
HPLC analysis was used for the determination of the purity of the administration solutions and for the determination of metabolites in urine and feces. The retention time of parent compound was determined for comparison with chromatograms of faeces samples. Analyses were performed with a HPLC-system consisting of a gradient pump (Merck-Hitachi L-6200), an autosampler (Merck-Hitachi AS-2000 A), an UV-detector (Merck-Hitachi L-4000), a radiodetector and a data processing system (Packard FLOW-ONE, Beta A 500).
For details on the methods used, please refer to Table 1 under "Any other information on materials and methods incl. tables".

Statistics:

The concentration of radioactivity in blood and plasma were expressed as parent equivalents on a fresh weight basis (pg eq/mL or g), and compared with the limits of quantification. Limits of detection (LOD) and limits of quantification (LOQ) for liquid scintillation counting were calculated according to L.A. Currie (Anal. Chem. 40, 586 (1968)) from the specific radioactivity of the test item.

Results and discussion

Main ADME results

Main ADME results 1

Type:

absorption

Results:

Both the MOH and the TBPH label were completely recovered in excreta, organs, tissues and carcass and in the cage wash. The mass balance in all groups ranged between 91.7-101%.

Main ADME results 2

Type:

distribution

Results:

Residues in the tissues at sacrifice were generally low. With the MOH label, the highest concentrations were found in the liver & kidney. Residues of the TBPH label were primarily found in adrenal glands, thyroid gland, liver, fatty tissue & ovaries.

Main ADME results 3

Type:

excretion

Results:

Both labels of the test item were rapidly excreted. 91-100% of the applied dose was recovered in urine and faeces within the first 48 h after administration. The major route of excretion was via the faeces.

Main ADME results 4

Type:

metabolism

Results:

15 metabolites detected in faeces (5 attr to unchanged parent/references, 10 unknown). 10 mg/kg bw (MOH/TBPH): the major metabolite was attr to ref.compound NP-2018 (45-71%). 1000 mg/kg bw: the major metabolite was attr to unchanged parent (62-65%).

Toxicokinetic / pharmacokinetic studies

Details on absorption:

The mass balance in all test groups ranged from 91.7 - 101% (refer to Table 2 under "Any other information on results incl. tables"). As the main excretion route was via the faeces (>90% of the dose for all test groups), the extent of oral absorption could not be determined.

Details on distribution in tissues:

In males and females of both labels and in all dose groups, the major part of the administered dose was excreted via the faeces. At the time of sacrifice, 0.065 - 0.27% (males) and 0.03 – 0.195% (females) of the dose were found in the organs, tissues, carcass and the content of the gastrointestinal tract. After administration of 10 mg/kg bw MOH-labelled test item, the highest concentrations found were in the kidneys and in the liver.
After administration of 10 or 1000 mg/kg bw TBPH-labelled test item, the highest amounts were found in the adrenal glands, liver, thyroids, ovaries and in fat. There were no marked differences found between males and females and between the low and the high dose group. For details, please refer to Table 3 under "Any other information on results incl. tables".

Details on excretion:

For both labels and both dose groups, the major excretion route was via the faeces (90.5 - 101% of the dose). There was no difference between males and females. Urinary excretion accounted for 1.14 – 1.77% of the dose for the MOH label and 0.05% or less for the TBPH label. More than 90% of the dose was excreted within 48 h post dosing. For details, please refer to Table 2 under "Any other information on results incl. tables".

Metabolite characterisation studies

Metabolites identified:

yes

Details on metabolites:

Metabolites were identified in the faeces (all treatment groups) and in the urine (MOH-labelled treatment group only).
In the faeces, 87 – 115% of the administered dose were recovered (refer to Table 4 under “Any other information on results incl. tables”). A total of 15 metabolite fractions were identified, 10 of them with unknown origin. Within the low dose groups, about 3.6 – 6.3% (groups 3 and 4 with 10 mg/kg bw MOH-labelled substance) and 17-23% (groups 7 and 8 with 10 mg/kg bw TBPH-labelled substance) of the parent compound were identified in the faeces. The main metabolite in the low dose group of both labels was F4, attributed to reference substance NP-2018.
In the high dose group (groups 5 and 6 with 1000 mg/kg bw TBPH-labelled substance), 62-65% of the radioactivity were attributed to unchanged parent substance and the main metabolite F4 accounted for 12-14% of the dose. The MOH label (groups 3 and 4) formed a more complex metabolite pattern as compared to the TBPH label. The relatively polar fractions F10-F15 were only observed with the MOH label, only F11 (2.4 – 3.6%), attributed to MOH, was more abundant than 1% of the dose. For details, refer to Table 6 under “Any other information on results incl. tables”.

In the urine samples, only the MOH-labelled test item groups (3 and 4) contained sufficient radioactivity for investigation. Recoveries were 88 – 102% (refer to Table 5 under “Any other information on results incl. tables”). Only two polar fractions were identified, with shorter retention times than any of the reference compounds. Parent and reference compounds were not found. For details, refer to Table 7 under “Any other information on results incl. tables”.

Any other information on results incl. tables

Table 2: Mean mass balance after a single oral administration of radiolabelled test material

Label		MOH				TBPH
Dose group (mg/kg bw)		10				10				1000
Sex		male		female		male		female		male		female
Test group		3		4		7		8		5		6
Compartment	Interval (h)	Mean	SD	Mean	SD	Mean	SD	Mean	SD	Mean	SD	Mean	SD
Urine	0-8	0.432	0.136	0.327	0.116	0.002	0.000	0.001	0.000	0.015	0.008	0.001	0.000
	8-24	1.060	0.250	0.650	0.248	0.005	0.002	0.045	0.086	0.006	0.005	0.002	0.002
	24-48	0.189	0.062	0.103	0.044	0.002	0.001	0.001	0.000	0.004	0.002	0.001	0.000
	48-72	0.041	0.012	0.025	0.010	0.001	0 .000	<0.001	-	0.003	0.003	0.001	0.000
	72-96	0.021	0.004	0.013	0.004	<0.001	-	<0.001	-	0.006	0.003	0.001	-
	96-120	0.012	0.002	0.009	0.002	<0.001	-	<0.001	-	0.001	0.001	0.001	-
	120-144	0.009	0.002	0.006	0.002	<0.001	-	<0.001	-	0.003	0.003	0.001	-
	144-168	0.007	0.001	0.005	0.001	<0.001	-	<0.001	-	0.002	0.001	<0.001	-
Subtotal		1.770	0.370	1.140	0.420	0.010	0.004	0.047	0.087	0.040	0.025	0.006	0.003
Faeces	0-24	89.2	3.3	83.1	5.2	89.2	5.9	90.5	3.6	90.6	2.0	83.2	9.7
	24-48	3.16	1.67	6.89	3.30	7.10	4.39	4.71	1.74	8.89	2.10	13.50	9.20
	48-72	0.391	0.572	0.301	0.217	0.198	0.112	0.118	0.020	0.615	0.339	0.703	1.119
	72-96	0.068	0.053	0.196	0.328	0.046	0.020	0.038	0.019	0.169	0.173	0.027	0.034
	96-120	0.033	0.008	0.013	0.008	0.024	0.008	0.929	1.814	0.148	0.102	0.016	-
	120-144	0.021	0.007	0.006	0.002	0.018	0.006	0.018	0.005	0.086	0.062	0.008	-
	144-168	0.016	0.004	0.005	0.001	0.014	0.005	0.016	0.004	0.080	0.050	0.012	-
Subtotal		92.9	2.5	90.5	2.3	96.6	1.7	96.3	2.3	101.0	1.0	97.5	2.4
Cage wash		0.013	0.005	0.018	0.012	0.002	0.001	0.008	0.014	0.062	0.044	0.002	0.000
Total excreted		94.7	2.5	91.7	2.1	96.6	1.7	96.3	2.36	101.0	1.0	97.5	2.4
Liver		0.044	0.012	0.016	0.004	0.037	0.012	0.052	0.011	0.008	0.003	0.006	0.001
Kidney		0.014	0.005	0.010	0.003	0.002	0.001	0.002	0.001	<0.005	-	<0.001	-
Organs/tissues		0.059	0.016	0.027	0.007	0.056	0.017	0.075	0.017	0.007	0.005	0.006	0.001
Carcass		0.031	0.007	0.016	0.002	0.097	0.026	0.114	0.020	0.252	0.253	0.047	0.055
Intestinal tract contents		0.006	0.002	0.002	0.000	0.006	0.002	0.006	0.001	0.009	0.007	0.002	0.001
Subtotal		0.065	0.025	0.029	0.010	0.159	0.045	0.195	0.037	0.268	0.263	0.056	0.057
Total		94.8	2.5	91.7	2.1	96.7	1.7	96.5	2.4	101.0	1.0	97.5	2.4

Table 3: Distribution in tissues [µg eq/g]

Label	MOH				TBPH
Dose group (mg/kg bw)	10				10				1000
Sex	male		female		male		female		male		female
Test group	3		4		7		8		5		6
Tissue	Mean	SD	Mean	SD	Mean	SD	Mean	SD	Mean	SD	Mean	SD
Adrenal Glands	<0.036	-	<0.036	-	0.084	0.027	0.238	0.073	<3.461	-	7.370	5.290
Blood	0.008	-	<0.007	-	<0.007	-	0.008	-	<0.692	-	<0.692	-
Brain	<0.007	-	<0.007	-	<0.007	-	<0.007	-	<0.692	-	<0.692	-
Epididymes	<0.007	-	-	-	0.013	0.004	-	-	<0.692	-	-	-
Eyes	<0.007	-	-	-	<0.007	-	<0.007	-	<0.692	-	<0.692	-
Fat	0.011	-	0.009	-	0.031	0.006	0.033	0.009	0.867	0.059	1.580	1.140
Femur	<0.008	-	<0.008	-	0.008	-	0.009	0.002	<0.733	-	<0.733	-
Heart	<0.008	-	<0.008	-	0.013	0.003	0.019	0.007	<0.733	-	1.100	-
Kidney	0.165	0.045	0.130	0.042	0.018	0.006	0.026	0.007	<0.733	-	1.590	-
Large intestinal tract	<0.008	-	<0.008	-	0.013	0.002	0.014	0.003	<0.733	-	<0.733	-
Liver	0.089	0.021	0.035	0.008	0.072	0.025	0.117	0.032	1.380	0.689	2.13	2.04
Lung	<0.008	-	<0.008	-	0.020	0.009	0.031	0.009	0.769	-	2.000	-
Muscle	<0.007	-	<0.007	-	0.008	0.001	0.011	0.002	<0.692	-	<0.692	-
Ovaries	-	-	0.007	-	-	_	0.126	0.054	_	-	2.980	3.440
Pancreas	<0.007	-	<0.007	-	0.026	0.008	0.032	0.009	0.930	-	2.060	-
Pituitary gland	<0.072	-	<0.072	-	<0.070	-	<0.070	-	<6.921	-	<6.921	-
Plasma	<0.007	-	<0.007	-	<0.007	-	0.008	-	<0.692	-	<0.692	-
Prostat gland	<0.007	-	-	-	0.016	0.004	-	-	<0.692	-	-	-
Sciatic nerve	<0.144	-	<0.144	-	<0.141	-	<0.141	-	<13.843	-	<13.843	-
Skin backregion	<0.007	-	<0.007	-	0.011	0.001	0.016	0.002	<0.692	-	1.030	-
Small intestinal tract	0.011	0.002	0.008	0.000	0.023	0.006	0.024	0.008	<0.692	-	1.200	-
Spinal cord	<0.007	-	<0.007	-	<0.007	-	<0.007	-	<0.692	-	<0.692	-
Spleen	<0.008	-	<0.008	-	0.024	0.010	0.030	0.008	<0.779	-	2.120	-
Stomach	<0.007	-	<0.007	-	0.014	0.005	0.018	0.004	<0.692	-	1.300	-
Testes	<0.007	-	-	-	0.010	0.002	-	-	<0.692	-	-	-
Thymus	<0.007	-	<0.007	-	0.015	0.005	0.017	0.004	<0.692	-	0.928	-
Thyroid gland	0.030	0.008	0.030	0.004	0.038	0.013	0.045	0.017	3.640	0.880	3.430	1.160
Urinary bladder	<0.072	-	<0.072	-	<0.070	-	<0.070	-	<6.921	-	<6.921	-
Uterus	-	-	<0.072	-	-	-	<0.070	-	-	-	<6.921	-
< below lower limit of quantification

Table 4: Recovery of radioactivity from the faeces

Label	MOH				TBPH
Dose group (mg/kg bw)	10				10				1000
Sex	male		female		male		female		male		female
Test group	3		4		7		8		5		6
Tissue	% in faeces	% of dose	% in faeces	% of dose	% in faeces	% of dose	% in faeces	% of dose	% in faeces	% of dose	% in faeces	% of dose
0-24 h	92	82	87	73	85	75	88	73	93	83	88	73
24-48 h	97	3	115	8	94	5	103	7	94	5	103	7

Table 5: Recovery of radioactivity from the urine of rats administered 10 mg/kg bw test item labelled with MOH

Label	MOH
Dose group (mg/kg bw)	10
Sex	male		female
Test group	3		4
Tissue	% in urine	% of dose	% in urine	% of dose
0-8 h	88	0.38	100	0.32
8-24 h	92	0.98	102	0.67

Table 6: Metabolites identified in the faeces

Label			MOH		TBPH
Dose group (mg/kg bw)			10		10		1000
Sex			male	female	male	female	male	female
Test group			3	4	7	8	5	6
Fraction	Designation	Retention time	0-48 h	0-48 h	0-48 h	0-48 h	0-48 h	0-48 h
F1	Parent compound	23.2-25.0	3.6	6.3	17.0	23.3	65.3	61.6
F2		20.9-23.3	3.6	3.9	3.1	3.1	<0.05	0.4
F3		20.1-22.4	1.2	1.0	-	-	0.5	0.5
F4	NP-2018	18.8-21.1	70.6	63.9	51.8	45.3	14.1	11.8
F5		17.5-18.9	1.9	1.6	1.1	1.0	0.6	0.4
F6	NP-1907/TBPH	16.6-18.1	<0.05	-	2.0	1.2	1.2	1.0
F7	NP-2017	11.8-13.0	-	-	8.5	4.9	3.1	3.8
F8		11.8	-	-	0.4	-	-	-
F9		10.0-11.1	-	-	1.0	0.6	-	-
F10		8.1-9.1	0.1	0.1	-	-	-	-
F11	MOH	6.3-7.1	3.6	2.4	-	-	-	-
F12		4.9-5.5	0.1	<0.05	-	-	-	-
F13		4.6	<0.05	<0.05	-	-	-	-
F14		2.9	0.2	0.1	-	-	-	-
F15		2.4	<0.05	0.1	-	-	-	-
#: sum of 0 - 24 h and 24 - 48 h sampling

Table 7: Metabolites identified in the urine of rats administered 10 mg/kg bw test item labelled with MOH

Sex		male				female
Test group		3				4
Fraction	Retention time	% in sample		% of dose		% in sample		% of dose
		0-8 h	8-24 h	0-8 h	8-24 h	0-8 h	8-24 h	0-8 h	8-24 h
U2	2.5-2.6	58.6	68.0	0.2	0.7	83.9	62.9	0.3	0.4
U1	3.2-3.3	41.4	32.0	0.2	0.3	16.2	37.1	0.1	0.2