Quantitation of the main constituents of vanilla by reverse phase HPLC and ultra‐high‐pressure‐liquid‐chromatography with UV detection: Method validation and performance comparison

Abstract

Vanilla's main constituents, i. e., vanillin, para-hydroxybenzaldehyde, and their corresponding acids, can be easily quantified by RP LC with UV detection and external calibration. This paper describes two methods that were developed using HPLC and ultra-high-pressure LC (UHPLC), respectively, and validated according to the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). Both methods were highly specific, exhibited good linearities with high precision, and achieved good accuracies of quantitative results. The UHPLC method was more sensitive, five times shorter, and gave better peak resolutions than the HPLC alternative.

Procedure for the isolation of vanillin from vanilla extracts prior to isotopic authentication by quantitative 13C‐NMR

“…Their initial vanillin amount was determined after multistep ASE[3] and HPLC quantification. [15] Based on this amount, the vanillin recovery in the alcoholic extract was 93%, and the recoveries of isolation/purification steps were identical to those that were obtained when starting from the model mixture (Figure 1).…”

“…All solutions were quantified using the UHPLC method described elsewhere[15] after filtration on 25 mm PTFE syringe filters with a pore size of 0.45 µm (Teknokroma, Sant Cugat del Vallès, Spain). Overall detection was achieved in the range of 210–600 nm, and the quantitative determinations were achieved at 260 (vanillic and pHB acids) and 280 nm (vanillin and pHB).…”

Methoxybenzaldehydes in Plants:Insight to the Natural Resources, Isolation, Application and Biosynthesis

“…In this method, ACN/methanol (solvent A, 1 : 1 v/v) and water/acetic acid (solvent B, 99.8 : 0.2 v/v, pH 2.88) were used as mobile phase with a gradient elution 29 . Further, an ultra-high performance liquid chromatography (UHPLC) method was developed for rapid and better separation of vanillin from vanilla bean extract using a C 18 RP column (ACQUITY UPLC BEH C 18 , 50 mm  2.1 mm, 1.7 m particle size) 30 . Another rapid HPLC method for separation of vanillin was established, where Synergi Hydro-RP reverse phase column (4 , 250 mm  4.60 mm) was used as stationary phase and water/methanol (68 : 32, v/v) as a mobile phase with an isocratic elution 2 .…”

Recent developments in the HPLC separation of phenolic compounds

“…Cicchetti and Chaintreau 87 compared HPLC and UHPLC analyses of vanilla constituents. UHPLC analysis employed a 50 mm 1.7 μm C18 column, whereas a conventional 250 mm 5 μm C18 column was used in HPLC analysis.…”