Determination of Chemical Constituents Content in Bentong and Non-bentong Ginger Using Reversed-Phase High Liquid Chromatography (RP-HPLC) Method

Chong, Yee Chen (2019) Determination of Chemical Constituents Content in Bentong and Non-bentong Ginger Using Reversed-Phase High Liquid Chromatography (RP-HPLC) Method. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Ginger scientifically known as Zingiber officinale Roscoe. Gingerols were identified as the major pungent components in the fresh ginger rhizome and will be transformed into shogaols when heating or storage. In this research, conditions of reversed phase high performance liquid chromatography (RP-HPLC) were optimized for the separation and determination of some chemical constituents, namely, [6]-gingerol, [8]-gingerol [10]-gingerol, [6]-shogaol and [8]-shogaol in bentong and non-bentong ginger powder. The objectives of this study are i) to determine the chemical constituent’s content of bentong and non-bentong ginger with different planting approaches using RP-HPLC method, ii) to validate the reversed-phase high performance liquid chromatography (RP-HPLC) for the determination of chemical constituents content of bentong and non-bentong gingers. iii) to determine proximate composition of bentong ginger that cultivated for 5 months in TARUC using soilless plantation method. The optimum condition for HPLC was a Luna C18 column, gradient elution using mobile phase of acetonitrile, injection with a 10 μL fixed loop and ultra-pure water and ultraviolet (UV) detection at 282 nm. Proximate analysis was conducted to identify the characteristics of selected 5-months cultivated bentong ginger powder that harvested from TARUC. Under the optimum condition, five chemical constituents were separated for all the bentong and non-bentong gingers within 30 minutes as the following order: [6]-gingerol (RT= 11.9 minutes), [8]-gingerol (RT= 18.3 minutes), [10]-gingerol (RT= 24.7 minutes), [6]-shogaol (RT=19.7 minutes), and [8]-shogaol (RT= 26.5 minutes). Linearity, sensitivity (limit of detection), and precision were investigated. The proposed HPLC method showed good analytical performance such as wide linearity with R2>0.9, LOD (0.05-0.3 μg/mL) and high precision (RT: RSD <2%) and peak area (RSD <5%). Among five bentong gingers and one non-bentong ginger sample investigated, four chemical constituents were found to contain [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol. [8]-shogaol was found not detected among the samples. For EB, ET, EN, ENE, LN (5), LN (9) sample the concentrations of [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol were ranged from 3.03-7.27 mg/100g, 0.27-0.85 mg/100g, 0.86- 2.79 mg/100g and 0.37-1.68 mg/100, respectively. Moisture content, crude fat, crude protein, crude fibre and ash content of 5 months bentong ginger powder were 90.93%, 9.62%, 1.22%, 5.92% and 1.36% respectively. Four chemical constituents present in these samples were successfully determined and validated using RP-HPLC. Keywords: Ginger, Gingerols, Shogaols, HPLC, validation, Proximate analysis