Calcium 4-chloro-2-(5-hydroxy-3-methyl-1-(3-sulfonatophenyl)pyrazol-4-ylazo)-5-methylbenzenesulfonate

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: well performed study according to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Objective of study:

other: absorption, distribution, metabolism, excretion

Principles of method if other than guideline:

The purpose of the study was to analyse the content of the test item in rat plasma, samples obtained in the course of a one to two generation reproduction oral toxicity study (RTC study 46350, see 7.8.1)
Blood samples were collected on days 10 and 61 of the study for males and on days 5 and 15 post coitum for females. In addition, blood samples were collected from the pups (both sexes) on the last day of dosing at 1, 3, 6, 24 and 48 hours after dosing.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: bulk

Radiolabelling:

no

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy S.r.l., San Pietro al Natisone (UD)
- Age at study initiation: (P ) Males: 5 wks; Females: 8-9 wks at receipt
- Weight at study initiation: (P) Males: 134.1-159.3 g; Females: 200.6- 229.4 g at receipt
- Housing: In a limited access rodent facility. At arrival, the animals were housed up to 5 of one sex to a cage, after allocation, during the pre-pairing period, the animals were housed 4 of one sex to a cage, during the mating period, the rats were housed one male to one female, males were re-caged after mating as they were before mating, after mating, the females were transferred to individual breeding cages.

- Diet (e.g. ad libitum): laboratory rodent diet (4RF21, Mucedola S.r.l., Via G. Galilei 4, 20019 Settimo Milanese (MI), Italy) ad libitum
- Water (e.g. ad libitum): Drinking water ad libitum
- Acclimation period: approximately 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: carboxymethylcellulose 1% in water

Details on exposure:

see IUCLID chapter 7.8.1 or RTC study No 46350 "One to Two Generation Reproduction Oral Toxicity Study in Rats (Including Sperm Analysis)"

Duration and frequency of treatment / exposure:

see IUCLID chapter 7.8.1 or RTC study No 46350 "One to Two Generation Reproduction Oral Toxicity Study in Rats (Including Sperm Analysis)"

Doses / concentrations

Remarks:

Doses / Concentrations:
see IUCLID chapter 7.8.1 or RTC study No 46350 "One to Two Generation Reproduction Oral Toxicity Study in Rats (Including Sperm Analysis)"

No. of animals per sex per dose / concentration:

see IUCLID chapter 7.8.1 or RTC study No 46350 "One to Two Generation Reproduction Oral Toxicity Study in Rats (Including Sperm Analysis)"

Positive control reference chemical:

see IUCLID chapter 7.8.1 or RTC study No 46350 "One to Two Generation Reproduction Oral Toxicity Study in Rats (Including Sperm Analysis)"

Details on dosing and sampling:

Plasma samples of different rats treated with the test item were collected and stored at - 80°C until analysis.

Blood samples were collected from the tail vein of the first 5 high dose and 5 control male rats on days 10 and 61 of dosing, 4 h after administration.
For females, blood samples were collected from the tail vein of the first 5 high dose and 5 control rats, 4 h after administration on day 5 of dosing and during pregnancy (day 15 post coitum).
For the pups, blood samples were collected from the tail vein of the first 8 male and 8 female pups of the high dose group (week 5 of study) at 1, 3, 6, 24, 48 h after dosing (4 rats per sampling time; 2 or 3 blood samples from each animal).
Blood samples were transferred into tubes containing lithium heparin as anticoagulant and centrifuged at room temperature at 3000 rpm for 10 min. For each sample the content of the test item was determined.

Statistics:

Mean, standard deviation (SD) and coefficient of variation (CV) of the individual data of the test item were calculated and used for the assessment of toxicokinetic data at RTC S.p.A.
The toxicokinetic analysis was conducted according to a standard non-compartmental analysis.
The following toxicokinetic parameters were obtained or calculated from the mean plasma values instead of the individual plasma levels as specified in the study protocol:
Cmax: maximum observed plasma concentration
tmax: time to Cmax
AUC0-48 h: area under the concentration-time curve calculated by the linear trapezoidal rule.

All the toxicokinetic parameters were estimated or calculated by the Kinetica (TM), version 4.4.1, PK/PD Analysis (Thermo Electron Corporation Informatics, Philadelphia - USA) software


For statistical evaluation and assessment of toxicokinetic parameters all values below the limit of quantitation (LLOQ = 49.752 ng/mL) were set to 0 (zero).
For the AUC0-48h calculation, zero was set at a 24 hour time for the male pups and at a 48 hour time for the female pups. In addition, for female pups the mean from the two numerical values was set at 24 hours.

Results and discussion

Toxicokinetic / pharmacokinetic studies

Toxicokinetic parameters

Toxicokinetic parameters 1

Key result

Toxicokinetic parameters:

Cmax: 191.39 ng/mL; male pups, mean of 4 animals

Toxicokinetic parameters 2

Toxicokinetic parameters:

Tmax: 1 h after dosing; male pups

Toxicokinetic parameters 3

Key result

Toxicokinetic parameters:

Cmax: 169.66 ng/mL; female pups, mean of 4 animals

Toxicokinetic parameters 4

Toxicokinetic parameters:

Tmax: 1 h after dosing; female pups

Metabolite characterisation studies

Metabolites identified:

no

Bioaccessibility (or Bioavailability)

Bioaccessibility (or Bioavailability) testing results:

Detectable plasma concentrations of the test item were found in male and female pups from 1 hour after dosing. The test item was below the limit of quantitation (LLOQ = 49.752 ng/mL) in 3 out of 4 male pups at 24 h after dosing and in all male pups at 38 h after dosing. For female pups the test item was below LLOQ in 2 out of 4 at 24 h after dosing and in 3 out of 4 at 48 h after dosing. The maximum plasma concentration of the test item was found 1 hour after dosing in pubs of both sexes.

For male and female rats the test item was detectable 4 hours after dosing on days 5 and 15 (females) and on days 10 and 61 (males)

Any other information on results incl. tables

Table 1: individual plasma concentrations of the test item in male pups collected on the last day of dosing (1000 mg/kg bw/day)

Animal No	Pub sex	Sampling Time [hours after dosing]	Test item concentration [ng/mL]
66340386	male	1	219.49
66340388	male	1	158.63
66340390	male	1	245.57
66340392	male	1	141.88
mean			191.39
SD			49.16
CV %			25.69
66340394	male	3	111.50
66340396	male	3	232.35
66340398	male	3	169.05
66340400	male	3	147.04
mean			164.98
SD			50.78
CV %			30.78
66340386	male	6	127.19
66340388	male	6	105.85
66340390	male	6	269.95
66340392	male	6	97.30
mean			150.07
SD			80.90
CV %			53.91
66340394	male	24	BLOQ
66340396	male	24	BLOQ
66340398	male	24	BLOQ
66340400	male	24	98.26
mean			24.57
SD			49.13
CV %			200.00
66340386	male	48	BLOQ
66340388	male	48	BLOQ
66340390	male	48	BLOQ
66340392	male	48	BLOQ
mean			0
SD			0
CV %			0

 

Table 2: individual plasma concentrations of the test item in female pups collected on the last day of dosing (1000 mg/kg bw/day)

Animal No	Pub sex	Sampling Time [hours after dosing	Test item concentration [ng/mL]
66340385	female	1	144.16
66340387	female	1	185.17
66340389	female	1	179.39
66340391	female	1	169.92
mean			169.66
SD			18.13
CV %			10.68
66340393	female	3	99.80
66340395	female	3	75.78
66340397	female	3	86.23
66340399	female	3	95.89
mean			89.43
SD			10.74
CV %			12.00
66340385	female	6	182.98
66340387	female	6	59.13
66340389	female	6	82.36
66340391	female	6	192.88
mean			129.36
SD			68.35
CV %			52.84
66340393	female	24	94.42
66340395	female	24	BLOQ
66340397	female	24	66.43
66340399	female	24	BLOQ
mean			40.71
SD			48.58
CV %			119.32
66340385	female	48	BLOQ
66340387	female	48	BLOQ
66340389	female	48	BLOQ
66340391	female	48	113.24
mean			28.31
SD			56.62
CV %			200.00