Development of High-performance Thin-layer Chromatography


High-Performance Thin Layer Chromatography (HPTLC) is a reliable and rapid method useful for the detection of product quality and composition. In recent times, HPTLC instrumentation allows to obtain fingerprints suitable to determine composition and identity of plant raw materials. This study aimed at exploring the results of High-performance Thin Layer Chromatography (HPTLC) analysis as a useful means for the authentication of Allanblackia parviflora kernel and seed oils using the fingerprint approach. Sixteen bulked kernel and seed oil samples from 16 different communities were analyzed by HPTLC method and their fingerprints were compared. The optimum experimental conditions established were sample mass of 20 mg, hexane as appropriate dissolving solvent, toluene-ethyl acetate (85:15, v/v) as mobile phase solvent and p-anisaldehyde sulphuric acid as derivatization reagent. Using HPTLC fingerprinting, it was observed that no differences existed in fingerprints for all kernel and seed oil samples under illumination mode at 366 nm after derivatization. However, the seed oils showed clear, sharp and more intense bands indicating the presence of more phytochemicals. This suggests that there are phytochemicals in the seed shells/husks soluble in the seed oil.

Several plants from Lamiaceae family are used in the Saudi Arabia as a condiment and food preparation and are generally used in the traditional preparation to treat various diseases, including anti-inflammatory, antioxidant, and microbial infections. Some of Lamiaceae species such as Mentha longifolia, Rosemarinus officinalis and Salvia officinalis having pharmacological active compounds such as α-pinene and eucalyptol. The aim of present study was to develop an accurate and precise chromatographic technique for quantification of α-pinene and eucalyptol in the Lamiaceae plants.

The high-performance thin layer chromatography (HPTLC) method was developed as per International Conference on Harmonization (ICH) guideline. Simultaneous determination of α-pinene and eucalyptol was achieved by developing a densitometric analysis of high-performance thin layer chromatography (HPTLC). Silica gel 60 F254 glass-backed plates (E-Merck, Germany, 0.2 mm layers) as stationary phase and mixture n-hexane: ethyl acetate 8 : 2 (%, v/v) as mobile phase were used to produce a sharp, symmetrical and well-resolved peak at an Rf value of 0.19 ± 0.02 and 0.52 ± 0.04 for α-pinene and eucalyptol, respectively. Linearly range for α-pinene was 100–700 ng/spot (r2 = 0.9988), whereas that for eucalyptol was 1000–7000 ng/spot (r2 = 0.9987).

The developed method was found to be a simple, accurate, and precise, and it may be used to simultaneously analyses of many medicinal plants’ samples containing α-pinene and eucalyptol.

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Journal of Developing Drugs