Analysis of mannitol in pharmaceutical formulations using hydrophilic interaction liquid chromatography with evaporative light-scattering detection

Abstract

This study demonstrates the use of hydrophilic interaction liquid chromatography (HILIC) for the separation of both active and inactive ingredients in pharmaceuticals from a single injection. Excipients commonly used in parenteral formulations were separated using a gradient method employing increasing aqueous composition. An evaporative light-scattering detector (ELSD) provided direct detection of inactive excipients and inorganic salts lacking UV chromophores. Analyses of Gemzar parenteral formulations using optimized isocratic HILIC-ELSD method conditions were performed based on retention time screening from the gradient assay. All of the components were efficiently separated using a TSK-Gel Amide 80 column including gemcitabine, mannitol, and sodium cation demonstrating the qualitative capability of the technique. The method was thoroughly validated for mannitol content to access the quantitative potential of the technique. Validation parameters included linearity, accuracy, specificity, solution stability, repeatability, and intermediate precision. Overall, the method described in this report proved to be very robust and represents a novel technique to conveniently separate and detect the active and inactive components in pharmaceuticals both quickly and accurately.

Hydrophilic interaction chromatography

“…Certain solutes seem to be more affected than others. [54,233,266]; pyridylaminated sialo sugar chains [219]; quaternary ammonium compounds [133]; small polar compounds in food analysis by HILIC/ESI-MS [155]; hydroxybenzoate saxitoxin analogs in dinoflagellates [160]; chitine fragments [198]; cyanobacterial toxins by HILIC/MS [164]; 1-deoxynojirimycin in mulberry leaves [150]; polar compounds contributing to umami taste and mouth-drying oral sensation of morel mushrooms [156]; elucidation of yeast sulfur metabolism [253]; PSP toxins [166,165,167]; monosaccharides [227,267]; identification of a bitter inhibitor [157] Peptides, nucleic acids, and other polar compounds [13]; peptides [67]; tyrosine protein-kinase specificity [234]; desalting of electroeluted proteins [241]; glycopeptides [268]; site-specific glycosylation microheterogeneity in recombinant human interferon-c [269]; urea, allantoin and lysine pyroglutamate in cosmetic samples [134]; glucosinolates in vegetables [201]; folates in human plasma by HILIC/ MS/MS [137]; citrulline in dried blood spots [138]; SPE enrichment of avoparcin from kidney samples [254]: Ionic compounds by pressurized flow CEC [270]; sucrose in a plasma-derived process solution [271]; glycoalkaloids in transgenic potatoes [104]; glycopeptide microheterogeneity [272] Peptides [67,68,276]; amphipathic a-helical peptides …”

Method development for HILIC assays

“…Separations of monosaccharides (L-fucose, D-galactose, D-mannose, N-acetyl-D-glucosamine, N-acetylneuraminic acid, D-glucuronic acid) common in N-linked oligosaccharides in glycoproteins [28], gemcitabine, mannitol, and sodium cation in pharmaceutical formulations [30], spectinomycin and lincomycin in liquid hog manure and run-off waters from manure-treated cropland [32], 2-aminopyridine derivatized N-glycans and N-glycopeptides of human serum immunoglobulin [34], a fine particle dose of lactose in a pharmaceutical dry inhalation product [37], and 2-aminopyridine derivatized sialylated N-glycans and N-glycopeptides of a-1-acid glycoprotein [38], were also determined with HILIC.…”

Simultaneous determination of nucleosides, myriocin, and carbohydrates in Cordyceps by HPLC coupled with diode array detection and evaporative light scattering detection

“…Sterols, especially ergosterol, have also been determined using HPLC-DAD [5], LC-MS [10] and GC-MS [11]. However, a few reports [12][13][14] focus on the carbohydrates such as mannitol, a characteristic component in Cordyceps, which have diuretic, decrease neutrophil apoptosis and antioxidant activities [15][16][17].…”

“…Therefore, ELSD is increasingly being used in LC as a quasiuniversal detector, which has been successfully applied to analyze carbohydrates in traditional Chinese medicines such as Picrorhiza [29], Chinese artichoke [30] or in pharmaceutical fomulations [13] and drinks [31]. ELSD can be [32][33][34].…”