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EC number: 439-210-6
CAS number: 554-62-1
The mobile phase was 60/40 (v/v) acetonitrile/Milli-Q water, and a
refractive index detector was used. The column oven temperature was set
to 22.0°C during the test. HPLC chromatograms of the Phytosphingosine
test solution, the test substance blank solution (methanol) and the
2,4-DDT reference solution are provided. In the chromatograms of the
1840 mg/L Phytosphingosine test solution, which were recorded for 90
minutes, no test substance peak was observed. Under the same
chromatographic conditions, the retention time of 2,4-DDT (log P. = 6.2)
was 57.2 minutes. It was assumed that Phytosphingosine elutes after the
reference substance 2,4 -DDT. The column oven temperature was set to
22.0°C during the test.
In order to show that the test substance can be passed through the HPLC
column and can be detected using a refractive index detector, a second
mobile phase (60/40/0.05 (v/v/v) acetonitrile/Milli-Q
water/trifluoracetic acid) was used. Methanol, the 2,4 -DDT solution,
the test solution (in duplicate), the 2,4-DDT solution and methanol were
injected using this mobile phase. One large test substance peak at a
retention time of 3.49 minutes was observed in the chromatograms of the
1836 mg/L Phytosphingosine test solution. Therefore, it was concluded
that at neutral pH, Phytosphingosine elutes
after the reference substance with known log P of 6.2. Hence, the log
Pow of Phytosphingosine is > 6.2 (P > 1.6 x 10^6). Under the conditions
used, a non-ionised substance will elute later than an ionised one.
Because Phytosphingosine is ionised at neutral pH and non-ionised at pH
10.5, it was concluded that the test substance will not elute before the
reference substance and that log Pow of Phytosphingosine is also > 6.2
(P ow > 1.6 x 10^6) at pH 10.5.
The results of the Calculation method and the HPLC method were not in
agreement. Since the HPLC method is a more accurate method than the
Calculation method, the result of the HPLC method is reported as the
partition coefficient (n-octanol/water), Pow, of Phytosphingosine. The
partition coefficient (n-octanol/water), Pow, of Phytosphingosine is
>1.6 x 10^6 (log Pow > 6.2).
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