VALIDATION OF GC-MS METHOD FOR STANDARDIZATION OF CURCUMA XANTHORRHIZA EXTRACTS USING BIOCHEMICAL MARKERS, AR-CURCUMENE AND XANTHORRHIZOL
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Date
2014-08
Authors
AB HALIM, MOHD ROHAIMI
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Abstract
In this thesis, the chemical constituents from rhizomes of C. xanthorrhiza Roxb. were investigated. Extraction of rhizomes of C. xanthorrhizawas conducted using maceration and Soxhlet extraction method that gives three different extracts namely ethanol, aqueous and hexane extracts. Isolation and purification of chemical constituents were done on the C. xanthorrhiza hexane extract. Two pure compounds was successfully isolated and characterized as ar-curcumene and xanthorrhizol. These isolated compounds were chosen as the chemical markers in the standardization of C. xanthorrhiza extracts. A new Gas Chromatography Mass Spectrometry analytical method was developed and validated for the assay of chemical markers in C. xanthorrhiza extracts and its pharmaceutical products. The GC-MS in the single ion monitoring (SIM) mode gave short analysis time, sufficient limit of detection (LOD) and limit of quantification (LOQ) and good assay precision and accuracy. Ar-curcumene content in the ethanol and aqueous extract was 136.02 ± 5.11 μg mL-1 and 21.08 ± 0.10 μg mL-1, respectively. The xanthorrhizol content for ethanol extract was found to be 228.86 ± 16.10 μg mL-1and 34.09 ± 0.93 μg mL-1 for the aqueous extract. Identification of compounds using the validated method revealed the present of ar-curcumene, α-cedrene, β-elemenone, xanthorrhizol, camphor, zingiberene, γ-elemene, transβ-farnesene and benzofuran. Ar-curcumene and xanthorrhizol were found to be the major compounds in the extracts. Initial qualitative phytochemical screening showed that C. xanthorrhiza extracts contain terpenoids, phenols, saponins, flavonoids, cardiac glycosides, alkaloids and coumarins while anthraquinones, anthrones and tannins were absent. Quantitative analysis of the total phenol content (TPC) showed that ethanol extract (199.00 ± 1.31 mg GAE g-1) contained higherpolyphenolic compounds compared to aqueous extract (19.99 ± 0.16 mg GAE g-1). A similar pattern as TPC was observed for total flavonoid content (TFC) with ethanol extract showing a higher amount of TFC compared to the aqueous extract with values of 101.66 ± 0.83 mg CE g-1 and 10.58 ± 0.83 mg CE g-1, respectively. Total saponin content was 80.90 mg g-1 and total alkaloid content was14.06 mg g-1. The result from stability studies revealed that the marker compounds (ar-curcumene and xanthorrhizol) were pH sensitive and degraded rapidly at higher temperature but were slight stable under photochemical exposure. Profiling of chemical markers based on developed High Performance Liquid Chromatography method showed ar-curcumene and xanthorrhizol in ethanol exhibited acceptable absorption peaks at 270 nm with retention times of 9.03 min and 3.21 min, respectively.
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BIOCHEMICAL , XANTHORRHIZOL