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EC number: 202-903-7
CAS number: 100-94-7
Table 5: Peptide depletion of NC, PC and the test substance for
Reaction with cysteine-peptide
Peptide depletion [%]
PC: EGDMA in ACN
Table 6: Peptide depletion of NC, PC and the test substance for
Table 7: Mean peptide depletions of Cysteine, Lysine and both peptides
Mean of both depletions [%]
Mean depletion [%]
The reactivity of the test substance towards synthetic cysteine (C)- or
lysine (K)-containing peptides was evaluated in the Direct Peptide
Reactivity Assay (DPRA). For this purpose, the test substance was
incubated with synthetic peptides for ca. 24 hours at ca. 25°C and the
remaining non-depleted peptide concentrations were determined by high
performance liquid chromatography (HPLC) with gradient elution and
UV-detection at 220 nm.
The test substance was dissolved at 100 mM in acetonitrile. Three
samples of the test substance were incubated with each peptide in ratios
of 1:10 (for C-containing peptide) or 1:50 (for K-containing peptide).
Additionally, triplicates of the concurrent vehicle control (= VC) were
incubated with the peptides.
Further, in order to detect possible interference of the test substance
with the peptides, a coelution control was performed and the samples
were analyzed by measuring UV absorbance at 258 nm in order to calculate
the area ratio 220 nm / 258 nm.
The following results were obtained in the DPRA:
The test substance was dissolved in acetonitrile at a concentration of
100 mM. The samples of the test substance with the peptides were
solutions at the time of preparation. Visual observation after the
24-hour incubation time did not reveal precipitates in any samples of
the test substance with the peptides.
No co-elution of test substance and peptides was present.
The mean C-peptide depletion, caused by the test substance was
determined to be - 0.12 %.
The mean K-peptide depletion, caused by the test substance was
determined to be 0.08 %.
Negative depletions were considered to be “zero” for calculation of the
mean peptide depletion, which was thus calculated to be 0.04 %.
Based on the observed results and applying the cysteine 1:10 / lysine
1:50 prediction model it was concluded that the test substance shows
minimal or no chemical reactivity in the DPRA under the test conditions
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