Analytical methodologies applied to the D003, new active ingredient obtained from cane wax of sugar

Abstract

D003 is a new active ingredient made-up of a mixture of saturated
linear fatty acids ranging from 24 and 36 carbon atoms. This product has been
isolated and purified from sugar cane (Saccharum officinarum L.) wax and has
shown antiosteoporotic, antioxidant, antiplatelet, and cholesterol-lowering effects.
Aqueous suspensions for preclinical studies and 5 and 10 mg dose tablets using
this active ingredient have been elaborated. In that sense, the development and
validation of gas chromatographic analytical methodologies for their determination
has been required. These methodologies are based on the employment of a
DB-5 wide-bore column and 1-nonadecanoic acid as internal standard. The acids
were analyzed as methyl ester derivatives, prepared using 5 % aqueous HCl-methanol.
Validated methods were specific, because no interference was detected, even
when samples were subject to stress conditions. Good linearity (correlation coefficient
> 0.99) and accuracy (total average recovery between 99.6 and 100.3 %) were
proved over a range 50-150 % of the nominal concentration. Repeatability, intermediate
precision and reproducibility at the nominal dose (100 %) were < 2.1 %. Robustness
studies showed that quantification and resolution were only affected by
the injection volume increase and no other of the 7 evaluated operational changes
affected the results. The methods proved to be suitable for quality controls and
stability studies of the active ingredient, tablets and suspensions.