Toxicological Studies
The safety of ForsLean® for use as a dietary supplement was established by means of detailed toxicological evaluation of the product. Tests performed include acute, sub-acute oral toxicity, sub-chronic oral toxicity, and bacterial reverse mutation assay. The oral LD 50 was established from two independently performed studies to be greater than 2000 mg/kg in rats, and ForsLean® was found to be non-mutagenic.
a) Acute Oral Toxicity (LD50)1
Acute oral toxicity of Coleus forskohlii 10% was determined in Sprague Dawley rats. The test substance suspended in water was administered by oral route and the experimental animals were observed for 14 days for product-related symptoms. The test substance did not produce any signs of intoxication after dosing and all animals survived the study period of 14 days. The LD 50 value of the product in rats by oral route was found to be greater than 2000mg/kg body weight.
b) Sub-chronic Toxicity1
Sub-chronic oral toxicity study was designed and conducted to Determine the toxicity profile of Forslean® when administered daily for 28 days in Sprague Dawley rats. Forslean® suspended in 0.1% aqueous CMC was administered to animals at varied doses. Their body weight and biochemistry were monitored during the period of experimentation. Based on the findings the no observed effect level (NOEL) of Forslean® in these experimental animals over a period of 28 days was found to be 100 mg/kg for male and female animals.
c) Sub-Acute Oral Toxicity1
Sub-acute oral toxicity study was designed and conducted to determine the toxicity profile of Forslean® when administered daily for 28 days in Sprague Dawley rats. Test material suspended in 0.1% aqueous CMC was administered to the animals at various dose levels. Hematological and biochemical analyses were carried out at the end of experimentation. All the animals survived the experimentation period and did not present any signs of intoxication. No abnormalities were detected. Based on these findings it was concluded that the no observed effect level (NOEL) of this product administered to rats orally over a period of 28 days was found to be 1000 mg/kg body weight for both male and female animals.
d) Single Dose Oral Toxicity in rats/LD50 in rats2
Animal model studied:
Rats, n=10
Study duration : 14 days
Results
- LD 50
- The oral LD 50 of Forslean® is greater than 2000mg/kg.
- Mortality
- All animals survived the 2000mg/kg oral dose.
- Body Weights
- Body weight changes were normal in 7/10 animals. Three females lost weight during the second week of the study.
- Systemic Observations
- Physical signs included diarrhea, soiling of the anogenital area and wetness of the mouth and anogenital area.
- Necropsy Findings
- Necropsy results were normal in all animals.
e) Bacterial Reverse Mutation Assay with an Independent Repeat Assay. Coleus ForsLean extract (ForsLean®) 3
Test methodology:
- ForsLean® was tested in the Bacterial Reverse Mutation Assay with an Independent Repeat Assay using Salmonella typhimurium tester strains TA98, TA100, TA1535 and TA1537 and Escherichia coli tester strain WP2 uvrA in the presence and absence of Aroclor-induced rat liver S9.
- The assay was performed in two phases, using the plate incoropration method. The first phase, the preliminary toxicity assay, was used to establish the dose-range for the mutagenicity assay.
- The second phase, the mutagenicity assay, (initial and independent repeat assays), was used to determine the mutagenic potential of the test article.
In the preliminary toxicity assay, the maximum dose tested was 5000 mcg per plate; this dose was achieved using a concentration of 100 mg/ml and 50 uL plating aliquot.
The overall evaluation and dose ranges tested are as follows:
|
S9 Activation
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Overall Evaluation and Dose Range Tested ( m g/plate )
|
|||||||||
|
|
Low
|
High
|
Low
|
High
|
Low
|
High
|
Low
|
High
|
Low
|
High
|
|
None
|
negative
|
negative
|
negative
|
negative
|
negative
|
|||||
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
|
|
Rat
|
negative
|
negative
|
negative
|
negative
|
negative
|
|||||
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
25
|
5000
|
|
Conclusion
ForsLean® was found to be non-mutagenic in the Bacterial Reverse Mutation Assay with an Independent Repeat Assay.
1Performed at the Indian Institute of Toxicology, Pune , India
2 Performed at MB Research Laboratories Spinnerstown , PA 18968 . USA
3 Performed at BioReliance Corporation, Rockville MD , USA .
