Table 6.
Summary of in vivo genotoxicity studies evaluated in the present opinion
| Test material | Mutagenicity assay | Test object | Route | Dose | Reported result | Reference | Commentsa |
|---|---|---|---|---|---|---|---|
| Aloe vera extracts and/or gels | Micronucleus formation | ICR, male mouse bone marrow cells | Oral gavage/ | 1,250, 2,500, 5,000 mg/kg bw once daily on two consecutive days | Negative | Williams et al. (2010) |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed |
| Fructus sennae |
Mouse spot test (Induction of different mutations in target genes which control the pigmentation of the coat hairs in melanoblasts) |
NMRI mice melanoblasts in developing embryos | Oral gavage |
100 and 1,000 mg/kg bw (single administration) |
Negative | Heidemann et al. (1996) |
Insufficient reliability. No evidence or demonstration that target tissues were sufficiently exposed. The Mouse Spot Test is not currently used/recommended in genotoxicity testing for induction of gene mutation in vivo. The relevant OECD Guideline 484 has been permanently withdrawn on 2 April 2014 |
| Micronucleus formation | Wistar male and female rat bone marrow cells | Oral gavage |
15,00 mg/kg bw (single administration) |
Negative |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. A limited number of PCE in the micronucleus test (1,000 PCE/animal) and no evaluation of the ratio between polychromatic and normochromatic erythrocytes to assess bone marrow toxicity |
||
| Chromosome aberrations | Wistar male and female rat bone marrow cells | Oral gavage |
Up to 1,000 mg/kg bw (single administration) |
Negative |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. No bone marrow toxicity observed. Selection of the highest dose level (1,000 mg/kg bw) not consistent with the highest dose level used for the micronucleus test (1,500 mg/kg bw) |
||
| Senna extracts | Micronucleus formation | NMRI male and female mouse bone marrow cells | Oral gavage |
2,000 mg/kg bw (single administration) |
Negative | Mengs et al. (1999) |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. No bone marrow toxicity observed. Maximum plasma levels of aloe‐emodin and emodin, two components genotoxic in vitro were 0.065 μg/mL and below the limit of detection (LOD), respectively. Only 2,000 PCE/animal) were scored for evaluation of micronuclei induction |
| Sennosides | Micronucleus formation | Bone marrow of male and female NMRI mice | Oral gavage |
2,500 mg/kg bw (twice 24 h apart) |
Negative | Heidemann et al. (1996) |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. No bone marrow toxicity observed. A limited number of PCE (1,000 PCE/animal) was scored |
| Emodin |
Micronucleus formation |
NMRI male and female mouse bone marrow cells | Oral gavage |
2,000 mg/kg bw (single administration) |
Negative | Mengs et al. (1997) |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. No bone marrow toxicity observed. Only 2,000 PCE/animal were scored for evaluation of micronuclei induction. Only one dose level |
| Aloe‐emodin |
Mouse spot test (Induction of different mutations in target genes which control the pigmentation of the coat hairs in melanoblasts) |
NMRI mice melanoblasts in developing embryos | Oral gavage |
200 and 2,000 mg/kg bw (single administration) |
Negative | Heidemann et al. (1996) |
Insufficient reliability. No evidence or demonstration that target tissues were sufficiently exposed. The Mouse Spot Test is not currently used/recommended in genotoxicity testing for induction of gene mutation in vivo. The relevant OECD Guideline 484 has been permanently withdrawn on 2 April 2014 |
| Micronucleus formation | Wistar male and female rat bone marrow cells | Oral gavage |
1,500 mg/kg bw (single administration) |
Negative |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. A limited number of PCE (1,000 PCE/animal) were scored for evaluation of micronuclei induction. Only one dose level administered. No bone marrow toxicity observed |
||
| Chromosome aberrations | Wistar male and female rat bone marrow cells | Oral gavage |
200, 666 and 2,000 mg/kg bw (single administration) |
Negative |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. Only two dose level administered. No bone marrow toxicity observed |
||
| In vivo/vitro unscheduled DNA synthesis | Primary hepatocytes of Wistar rats | Oral gavage |
100 and 1,000 mg/kg bw (single administration) |
Negative |
Insufficient reliability. No evidence or demonstration that target tissue was sufficiently exposed. Only two dose level administered. Not recommended as an adequate in vivo follow‐up study in genotoxicity testing (EFSA, 2017) |
||
| DNA fragmentation | Colon and kidney cells of male OF1 mice | Oral gavage | 500, 1,000 and 2,000 mg/kg (twice 24 h apart) |
Positive (colon); Weakly positive (kidney) |
Nesslany et al. (2009) |
Reliable. Essentially compliant with the current OECD Guideline 489 ‘in vivo mammalian alkaline comet assay’ |
bw: body weight; PCE: polychromatic erythrocytes.
- Reliable.
- Limited reliability (e.g. if certain aspects are not in accordance with OECD guidelines or current standards and/or limited documentation).
- Insufficient reliability (e.g. if main aspects are not in accordance with any recognised guidelines (e.g. OECD) or current standards and/or inappropriate test system).
- Reliability cannot be evaluated (e.g. insufficient documentation, short abstract only, too few experimental details provided).