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Administrative data

other: in-vitro
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Short communication on sodium bisulphite toxicity to human neuroblastoma cells.
Reason / purpose:
reference to same study

Data source

Reference Type:
Toxicity of chloroprocaine and sodium bisulfite on human neuroblastoma cells.
Seravalli, E.; Lear, E.
Bibliographic source:
Anesth. Analg. 66, 954-958

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
Plates of human neuroblastoma cells were incubated with sodium bisulphite concentrations in the range from 0.08 to 0.8 mM at pH 7.4 for 3 or 20 hours. After the exposure, the medium was discarded and drug-free medium was added to the experimental and control cultures. The cultures were left undisturbed at 37˚C for 3, 6, and 9 days. Thereafter, the cultures were washed with saline and stained with Wright-Giemsa for cell counting. Cell multiplication was measured by the number of colonies that developed in each experimental and untreated culture.
GLP compliance:
not specified
Limit test:

Test material

Details on test material:
- Name of test material (as cited in study report): sodium bisulfite
- Molecular formula (if other than submission substance):NaHSO3
- Molecular weight (if other than submission substance): 104 g/mol
- Substance type: technical product
- Physical state: solid, white powder
- Other: sodium bisulfite was obtained from two commercial sources, hencefore identified as SB-1 and SB-2
No further details are given.

Test animals

other: not applicable, in-vitro test
other: not applicable, in-vitro test
Details on test animals and environmental conditions:
not applicable, in-vitro test

Administration / exposure

Route of administration:
other: not applicable, in-vitro test
other: not applicable, in-vitro test
Details on exposure:
- sodium bisulfite was diluted in serum-free medium (DMEM-vide supra)
- dilution were made as follows: SB concentrations ranging from 0.08 mM to 0.8 mM
- dilutions of all drugs were prepared immediately before addition to the cells
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
3 or 20 hours
Frequency of treatment:
not applicable
Doses / concentrations
Doses / Concentrations:
0.08 - 0.8 mM sodium bisulfite
nominal conc.
No. of animals per sex per dose:
not applicable, in-vitro test
Details on study design:
- Dose selection rationale: the concentrations were chosen within limits that approximate the concentrations achieved in clinical practice


Reduction of colony forming ability was determined by expressing the number of colonies in drug-treated cultures as a percentage of the number of colonies in its control culture. Inhibition was considered definite when it was ≥25 %. Student’s t-test (two-tailed) was used for statistical analysis. P values <0.05 were considered statistically significant.

Results and discussion

Results of examinations

Clinical signs:
not examined
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Clinical biochemistry findings:
not examined
Behaviour (functional findings):
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined

Effect levels

Dose descriptor:
Remarks on result:
not determinable
no NOAEL identified

Any other information on results incl. tables

- Sodium bisulphite reduced the cell multiplication in neuroblastoma cells.

- The colony-forming ability (CFA) was inhibited by 72% to 92% at a 3-h exposure to one commercial sample of sodium bisulphite and by 57% to 72% with another sample (both tested at concentrations of 0.8 mM).

- no difference in the inhibition of CFA was detected between both commercial samples at concentrations below 0.8 mM

- Prolongation of exposure time to 20 hours resulted in a CFA inhibition of 98% by both samples.

- cells exposed to either sample of sodium bisulfite for 20 hr did not resume ability to form colonies during the study period of 9 days

- The authors concluded that sodium bisulphite may have neurotoxicity of potential clinical significance because it is used as antioxidant in commercial solutions of chloroprocaine.

Applicant's summary and conclusion

In this study sodium bisulfite was found to have an inhibitory effect on CFA of neuroblastoma that varied with concnetration and exposure time. Cell multiplication in human neuroblastoma cells was reduced by sodium bisulfite treatment (0.08-0.8 mM) for 3 or 20 hours.