Alfalfa, ext.

Acute Toxicity: oral

Administrative data

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

Data source

Reference

Reference Type:

publication

Title:

Assessment of beneficial and possible toxic effects of two new alfalfa-derived shelf products.

Author:

Soto-Zarazua MG et al.,

Year:

2016

Bibliographic source:

Journal of Medicinal Food, 19 (10), 970-977

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

not applicable - High molecular weight constituents likely to include lignins, proteins, polysaccharides and other natural polymer present in Alfalfa leaf

Molecular formula:

not applicable - High molecular weight constituents likely to include lignins, proteins, polysaccharides and other natural polymer present in Alfalfa leaf

IUPAC Name:

not applicable - High molecular weight constituents likely to include lignins, proteins, polysaccharides and other natural polymer present in Alfalfa leaf

Constituent 2

Reference substance name:

not applicable - Residue on ignition (including K, Mg and Ca) of alfalfa leaves

Molecular formula:

not applicable - Residue on ignition (including K, Mg and Ca) of alfalfa leaves

IUPAC Name:

not applicable - Residue on ignition (including K, Mg and Ca) of alfalfa leaves

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

Test animals

Species:

rat

Strain:

Wistar

Sex:

not specified

Details on test animals or test system 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

Administration / exposure

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.

Results and discussion

Effect levels

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:

other: Not specified

Gross pathology:

Not specified

Applicant's summary and conclusion

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.