Spinflam MF 83 PP-25

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is well described and scientifically acceptable. Justification for read across approach is explained in the endpoint summary.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1973

Materials and methods

Objective of study:

excretion

Principles of method if other than guideline:

The aim of this study was to compare the amount of piperazine excreted in the urine over 24 hrs after oral administration of a newly developed syrup formula, equivalent to 3.5 g. piperazine hexahydrate, against a commercial syrup sample. This dose was chosen according to the normal adult dose range for the treatment of ascariasis.

GLP compliance:

no

Remarks:

Pre GLP

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test animals

Species:

human

Strain:

other: Caucasian

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: ranging from 23 to 38 years of age.
- Health: normal state of health, determined by physical and laboratory medical examination.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: syrup

Duration and frequency of treatment / exposure:

Each experiment was repeated five times.

Doses / concentrations

Remarks:

Doses / Concentrations:
Each subject was administered an oral dosage of 35 ml of piperazine citrate syrup equivalent to 3.5 g. piperazine hexahydrate.

No. of animals per sex per dose / concentration:

Three men and two women.

Control animals:

other: Prior to the oral administration of either syrup sample, a urine sample was collected from each individual to be used as an internal control.

Details on study design:

In this work, a modified colorimetric method of Rogers* was used to achieve higher sensitivity and more accurate results. In Rogers' method, readings were taken at 480 nm. after standing for 10 min. The present work has shown that this length of time would allow neither for the optimum stability and intensity of the colour derivative nor for the reading at its maximum wavelength.

*Rogers E. W., Brit. Med. J., 1, 13 (1958).

DETERMINATION of PIPERAZINE in URINE
Piperazine was determined quantitatively in urine by its colour reaction with Folin's amino acid reagent (Sodium 1.2-naphthoquinone-4-sulfate).
The colour intensity increased rapidly from 0 to 18 min and was stable for 10 min. The maximum absorbance of the colour was determined by scanning the developed colour under visible light at a range of from 800 to 320 nm and was found to be at 490 nm.
The colour obeyed Beer's law in concentrations between 0 and 200 mcg/ml.
Each urine sample was measured for the exact volume and a dilution of 1 :25 with water was made for each sample including the control urine. This dilution was used for testing using the various reagents in the tabulating order (see the table below).
After standing for 20 min, readings were carried out within 5 min at 490 nm. The value of the unknown sample was then compared with the standard sample reading.
The validity of this method was verified by adding various known amounts of piperazine citrate to the urine. This urine was tested and assayed for the amount of piperazine present according to the described method.

CALCULATION of PIPERAZINE in URINE
The values obtained for the samples were corrected by subtracting the value of the normal urine reading for each individual. Piperazine concentration in the diluted urine sample was then calculated by comparing these corrected values with the standard reading carried out simultaneously.
The total piperazine concentration in the urine sample was calculated by multiplying its dilution factor and urine volume. The total amount of piperazine excreted and the percentage of excretion of each individual was then ascertained as piperazine hexahydrate.

Details on dosing and sampling:

Urine samples were collected, whenever possible, at 1, 2.5, 4.5. 6.5, 9, 13, 20, and 24 hrs after oral administration.

Results and discussion

Any other information on results incl. tables

Results of the total urinary excretion of piperazine calculated as hexahydrate after the oral administration of an amount equivalent to 3.5 g of piperazine hexahydrate

Subject (a)	Dose (b)	1 hrs	2.5 hrs	4.5 hr.	6.5 hrs	9 hrs	13 hrs	20 hrs	24 hrs	Total percentage excretion	t (c) Value
A	I	50	633	761	528	429	127	(d)	(d)	72	0.0982
27, 110	II	76	682	927	458	412	109	(d)	(d)	75
B	I	156	479	604	305	216	356	347	(d)	70
33, 73	II	196	176	530	340	275	391	353	77	75
C	I	153	562	647	372	261	177	50	18	64
38, 79	II	66	596	588	318	306	220	70	27	63
D	I	103	502	688	293	223	158	65	41	59
23, 57	II	46	334	582	413	486	104	73	61	60
E	I	54	263	160	30	(d)	(d)	(d)	(d)	15
29, 56	II	96	127	219	106	(d)	(d)	(d)	(d)	15

(a) The first number is for age (years) and the second number is for body weight (kilograms).

(b) I = new syrup formula, 3.5 g. piperazine hexahydrate. II = commercial syrup sample, 3.5 g piperazine hexahydrate.

(c) The r test of significance, where t = mean1 - (mean2/S) √ (n1n2/n1 + n2); p = 0.05, and df = 8 [“Remington’s Pharmaceutical Sciences,” 14th ed., Mack Publishing Co., Easton, Pa., 1970, p. 129]. The t value was calculated with the pooled variance of the two syrup samples.

(d) Zero excretion.

The percentage and pattern of urinary excretion of piperazine calculated as hexahydrate varied from individual to individual, as seen in the five subjects. The total urinary recovery varied from 15 to 75 %. However, the results and pattern of urinary excretion for both syrup formulas compared favorably within each individual.

Urinary excretion began 1 hrs after the oral administration, and the rate was maximal between 2 and 6 hrs. Excretion was nearly completed in 24 hrs.

CONCLUSION

1. The modified colorimetric method of analysis of piperazine in urine is accurate and simple to perform.

2. The percentage and pattern of urinary excretion of piperazine varied from individual to individual.

3. The percentage and pattern of urinary excretion after oral administration of the newly developed syrup formula, equivalent to 3.5 g. piperazine hexahydrate, compared favourably with that of the commercial syrup sample with each individual.

4. The maximum urinary excretion occurred 2-6 hrs after oral administration and was nearly completed at 24 hrs for both syrup formulas under investigation.

5. The t test of significance, where the t-value was calculated with the pooled variance of the two syrup samples, showed no significant difference.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): bioaccumulation potential cannot be judged based on study results
Within 24 hours between 60 to 75 % of the administered dose was excreted and the total recovery in urine collected during 24 hours varied from 15 to 75%.

Executive summary:

A modified colorimetric method using Folin's amino acid reagent was used for the quantitative determination of piperazine in human urine.

The aim of this study was to compare the amount of piperazine excreted in the urine over 24 hrs after oral administration of a newly developed syrup formula, equivalent to 3.5 g. piperazine hexahydrate, against a commercial syrup sample. This dose was chosen according to the normal adult dose range for the treatment of ascariasis.

The modified colorimetric method of analysis of piperazine in urine resulted accurate and simple to perform.

The percentage and pattern of urinary excretion after oral administration of the newly developed syrup formula, equivalent to 3.5 g. piperazine hexahydrate, compared favourably with that of the commercial syrup sample with each individual.

The percentage and pattern of urinary excretion of piperazine varied from individual to individual, nevertheless the maximum urinary excretion occurred 2-6 hrs after oral administration and was nearly completed at 24 hrs for both syrup formulas under investigation.

Conclusion

Within 24 hours between 60 to 75 % of the administered dose was excreted and the total recovery in urine collected during 24 hours varied from 15 to 75%.