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Description of key information

LD50 (oral) >3000 mg/kg bw (alfalfa-derived products oral suppplements)

LD50 (oral) >2250 mg/kg bw (phyto-preparation of alfalfa extract)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
no guideline followed
Principles of method if other than guideline:
Seven experimental groups (n = 3) were assigned randomly. The alfalfa derived products were orally administered at 1500 and 3000 mg/kg at a single administration in three independent experiments and one group was used as control (treated only with water). The survival rate was measured 24 and 48 h after treatment and results were reported as percentage of survival.
GLP compliance:
not specified
Remarks:
Study was performed in a research laboratory, probably none GLP compliant.
Test type:
standard acute method
Specific details on test material used for the study:
Test material
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Alfalfa-derived products for human consumption 1) freeze-dried juice obtained from fresh aerial parts by squeezing (FDJ), 2) dried and powdered residual material (RM). Both were manufactured from two different harvests, one in May (B1) and the other in August (B2) 2014.
- Expiration date of the lot/batch: 10.09.2019
- Purity test date: not available

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: not specified
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: stable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not available

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
- Preliminary purification step (if any): none
- Final dilution of a dissolved solid, stock liquid or gel: none
- Final preparation of a solid: not applicable

OTHER SPECIFICS: none

Species:
rat
Strain:
Wistar
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: not specified
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: not specified
- Weight at study initiation: 250-300 g
- Fasting period before study: not specified
- Housing: standard conditions as outlined in the Mexican Official Standard NOM-062-ZOO-1999
- Diet: ad libitum:
- Water: ad libitum
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
As outlined in the Mexican Official Standard NOM-062-ZOO-1999
Route of administration:
oral: gavage
Doses:
1500 and 3000 mg/kg as a single administration
Control animals:
yes
Details on study design:
Seven experimental groups (n = 3) were assigned randomly. The FDJ B1 and B2, and RM B1 and B2 were orally administered at 1500 and 3000 mg/kg at a single administration in three independent experiments. One group was used as control (treated only with water). The survival rate was measured 24 and 48 h after treatment and results were reported as percentage of survival.
Statistics:
Results were analyzed using Minitab 15. One-way analysis of variance (ANOVA) and two-way ANOVA were used to determine the difference between treatments. Post hoc comparisons of the means were performed according to Tukey and Bonferroni tests depending on the experiments. P < .05 was considered significant.
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 3 000 mg/kg bw
Mortality:
No altterations on survival were observed with 1500 mg/kg body weight for both FDJ and RM from both harvest times (B1 and B2).
88% survival rate was found when 3000 mg/kg body weight of FDJ from B1 harvest time was administered.
Clinical signs:
Not specified
Body weight:
Not specified
Gross pathology:
Not specified
Interpretation of results:
GHS criteria not met
Conclusions:
In an acute oral toxicity study in Wistar rats, a LD50 value for alfalfa-derived products was determined at >3000 mg/kg body weight.
Executive summary:

Adult Wistar rats were administered two different concentrations (1500 and 3000 mg/kg body weight) of two alfalfa-derived products (freeze-dried juice and dried and powdered residual material from 2 alfalfa harvest batches). Alfalfa-derived products were administered as a single dose in three independent experiments; and the control group animals received water. The survival rate was measured 24 and 48 hours after treatment. No alterations on survival were observed with 1500 mg/kg body weight dose after 24 and 48 hours. However, after 48 hours, only 88% of survival was found when 3000 mg/kg body weight of freeze-dried juice from one of the batches (B1) was administered. Based on the experiments, the alfalfa-derived products were considered of low acute toxicity under conditions of this experiment.

The value of LD50 in this study has been determined at >3000 mg/kg body weight, therefore, GHS criteria for acute oral toxicity were not met for alfalfa-derived products.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: study not well documented
Qualifier:
no guideline available
Principles of method if other than guideline:
Method poorly described.
GLP compliance:
not specified
Test type:
standard acute method
Specific details on test material used for the study:
Phytopreparation of Alfalfa (Medicago sativa) extract
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: gavage
Doses:
2250-7500 mg/kg body weight
No. of animals per sex per dose:
Not specified
Statistics:
Litchfield-Wilcoxon method
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 2 250 mg/kg bw
Mortality:
Not observed
Interpretation of results:
GHS criteria not met
Conclusions:
In an acute oral toxicity study in rats, a LD50 value for alfalfa extract phytopreparation was determined at >2250 mg/kg body weight.
Executive summary:

No symptoms of poisoning of death were observed when rats were administered a phyto-preparation of alfalfa extract at doses between 2250 and 7500 mg/kg body weight.

The value of LD50 in this study has been determined at >2250 mg/kg body weight, therefore, GHS criteria for acute oral toxicity were not met for alfalfa extract phytopreparation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
3 000 mg/kg bw

Additional information

Adult Wistar rats were administered two different concentrations (1500 and 3000 mg/kg body weight) of two alfalfa-derived products (freeze-dried juice and dried and powdered residual material from 2 alfalfa harvest batches B1/B2). Alfalfa-derived products were administered as a single dose in three independent experiments; and the control group animals received water. The survival rate was measured 24 and 48 hours after treatment. No alterations on survival were observed with 1500 mg/kg body weight dose after 24 and 48 hours. However, after 48 hours, only 88% of survival was found when 3000 mg/kg body weight of freeze-dried juice from one of the batches (B1) was administered. Based on the experiments, both alfalfa-derived products were considered of low acute toxicity under conditions of this experiment.

No symptoms of poisoning of death were observed when rats were administered a phyto-preparation of alfalfa extract at doses between 2250 and 7500 mg/kg body weight.

Additionally, there were no indications of adverse effects after administration of a single dose of 5000 mg/kg body weight of alfalfa protein concentrate (unpublished study cited in EFSA, 2009).

Justification for classification or non-classification

Appropriate tests were performed on the suitable analogues of Alfalfa, ext. to assess its potential to cause acute toxicity by oral route. The analogue substances were concidered of low acute toxicty under the conditions of the experiments (>2000 mg/kg bw/day), therefore, based on these results, it is concluded that Alfalfa, ext. does not meet the criteria for classification as acutely toxic by oral route in accordance with Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging.