Development and validation of a novel UPLC-ELSD method for the assessment of lipid composition of nanomedicine formulation

Abstract

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Quantification of Surface GalNAc Ligands Decorating Nanostructured Lipid Carriers by UPLC-ELSD

“…To analyze each component of the formulations, NLC were disassembled by precipitating the lipids in acetonitrile. Therefore, components of NLC were first separately analyzed by UPLC-ELSD according to previously described analytical methods [39]. Briefly, two UPLC-ELSD methods were previously developed for the analytical qualification of NLC, one using a reverse-phase (RP) C18 column (method A), separating and quantifying Myrj TM S40 and lipids, and the second one, a HILIC column (method B) for the assessment of lecithin components.…”

“…Briefly, two UPLC-ELSD methods were previously developed for the analytical qualification of NLC, one using a reverse-phase (RP) C18 column (method A), separating and quantifying Myrj TM S40 and lipids, and the second one, a HILIC column (method B) for the assessment of lecithin components. Analytical method A, which comprised a two-step gradient elution program, succeeded in analyzing the exact content of Myrj™S40 (three compounds), soybean oil (15 compounds), and Suppocire™NB (15 compounds) [39]. To specifically analyze additional SA-PEG 100 -GalNAc, the proposed RP-UPLC-ELSD method was herein optimized by extending the first gradient step (from 3 to 15 min), thus allowing better separation of Myrj™S40 and SA-PEG 100 -GalNAc.…”

“…According to the supplier information, Myrj TM S40 is a non-ionic surface active agent which was calculated to contain 71.3% ( w / w ) of three different compounds: PEG-C 18 with 18 carbon lipophilic chain (PEG-stearate), PEG-C 16 with 16 carbon lipophilic chain (PEG-palmitate) and PEG-OH without lipophilic chain [39]. They appeared in the chromatogram (Figure 1) as three peaks at 1.9, 6.4, and 7.2 min respectively.…”

“…It appeared in the chromatogram as 15 peak clusters in the range of 8 to 26 min, and elution occurred according to the equivalent carbon number. Super refined soybean oil TM was identified as being a mixture of C 16 –C 18 unsaturated triglycerides, and appeared in the chromatogram as eight peak clusters in the range of 17 to 24 min [39]. Lipoid TM s75 is a phospholipid and is then mainly composed of phosphatidylcholine with a slight proportion of phosphatidylethanolamine and lysophosphatidylcholine.…”

“…The peak areas of SA-PEG 100 -GalNAc 6 , PEG 40 -OH, PEG 40 -C 16 and PEG 40 -C 18 were plotted against surfactant concentration. As previously used, a second-order polynomial (quadratic) was used as a fitting model [39], producing an excellent correlation coefficient (R 2 values comprised between 0.9999 and 0.9994) (Figure 2, Figures S3 and S4).…”