Acute Toxicity Test for Eezapein^TM

(Maximum Tolerance Measurement)

Abstract:

 

Mice were administered Eezapein^TM extract four times a day until the maximum tolerance dosage (MTD) was determined to be 186.37 times the clinical adult dosage (crude drugs 180g/day).

 

Objective:

 

Since LD₅₀ could not be measured due to the low toxicity of Eezapein^TM; MTD was determined instead.

 

Materials:

 

1. Eezapein^TM, provided by LIFEnhance, Inc., is a brown extract, batch number 20020928. 1g extract is equivalent to 6g crude drugs. Extract is dissolved in distilled water to a final concentration of 93.18%, and stored at 4°C until use.

 

2.    Kunming mice were obtained from Hubei Animal Experiment Center.  Kunming mice weighed 18.5±0.5g, equal numbers ♂(male) and ♀(female).  The quality certificate number is 19-082. Mice feed also obtained from Hubei Animal Experiment Center.

 

Methods:

 

Adherent to established protocols [1], 24 Kunming mice (weight 18.5±0.5g) half ♂(male) and half♀(female), were starved for 16 hours, then fed 27.955g Eezapein^TM extract per dose (30ml/kg) four times a day.  Mice were inspected daily for 7 consecutive days.  In total 111.82g/kg (equivalent to 670.92g/kg crude drugs) of Eezapein^TM extract was administered; 186.37 times clinical dosage.

 

Result: 

No fatalities were observed nor were there any behavioral abnormalities.  Mice feces seemed softer and were of a slightly darker color.  After two dosages feed consumption in three ♀ declined somewhat but recovered by the following day.  Feed consumption decline was accompanied by loose stools in these mice.  Tolerance levels were calculated as follows:

 

                       Per mice’s tolerance   adult average weight (50,000g)

The mice’s tolerance times=—————————×—————————————

                       Average weight (20g)      adult’s daily dosage

 

                        0.559g×4   50000g

=————×————=186.367 times

                                                   20g     30g

 

i.e. The mice tolerance is 186.37 times adult dosage, which is far beyond 100 times.

The tolerance is the total amount of 27.955/Kg medicine x 3=111.82g/kg, equal to 670.92g/kg crude drugs.

 

Reference:

[1] Guidebook for New Medicine Research of Chinese Herbal Medicine. Medicine Management Department of Public Health Ministry of P.R China, P203-204

[2] Research Methodology of Pharmacology of Chinese Herbal Medicine by Chen Qi, People Health Press, P118-119

Testing Facility: Chinese and Western Medicine Institute of Hubei Traditional Chinese Medicine Academy

Studies conducted from October to December of 2002