Application of Scion image software to the simultaneous determination of curcuminoids in turmeric (<i>Curcuma longa</i>)

The eukaryotic initiation factor 3 (eIF3) is an essential, highly conserved multi-protein complex that is a key component in the recruitment and assembly of the translation initiation machinery. To better understand the molecular function of eIF3, we examined its composition and phosphorylation status in Saccharomyces cerevisiae. The yeast eIF3 complex contains five core components: Rpg1, Nip1, Prt1, Tif34, and Tif35. 2-D LC-MS/MS mass spectrometry analysis of affinity purified eIF3 complexes showed that several other initiation factors (Fun12, Tif5, Sui3, Pab1, Hcr1, and Sui1) and the casein kinase 2 complex (CK2) co-purify. In vivo metabolic labeling of proteins with 32P revealed that Nip1 is phosphorylated. Using 2-D LC-MS/MS analysis of eIF3 complexes, we identified Prt1 phosphopeptides indicating phosphorylation at S22 and T707 and a Tif5 phosphopeptide with phosphorylation at T191. Additionally, we used immobilized metal affinity chromatography (IMAC) to enrich for eIF3 phosphopeptides and tandem mass spectrometry to identify phosphorylated residues. We found that three CK2 consensus sequences in Nip1 are phosphorylated: S98, S99, and S103. Using in vitro kinase assays, we showed that CK2 phophorylates Nip1 and that a synthetic Nip1 peptide containing S98, S99, and S103 competitively inhibits the reaction. Replacement of these three Nip1 serines with alanines causes a slow growth phenotype.

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“…32 P labeled-proteins were detected by autoradiography. Densitometric measurements were quantified using Scion image software 48 .…”

Section: Methodsmentioning

confidence: 99%

Assessing the Components of the eIF3 Complex and their Phosphorylation Status

Farley¹,

Powell²,

Weaver³

et al. 2011

J. Proteome Res.

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“…Chromatographic methods are extensively used for quantitative analysis of curcuminoids and also for fingerprint analysis of Curcuma species. A number of qualitative and quantitative analysis of curcuminoids using thin layer chromatography (Sotanaphun et al 2009), HPLC with UV or fluorescence detector (Jadhav et al 2007;Wichitnithad et al 2009;Cheng et al 2010;Li et al 2011), liquid chromatography-mass spectrometry (LC-MS) (Verma et al 2013), micellar electrokinetic chromatography (Watanabe et al 2000), and capillary electrophoresis (Lechtenberg et al 2004;Anubala et al 2014) have been reported in various Curcuma samples. In the chromatographic fingerprint analysis of Curcuma species, the reported analytical instruments used are HPLC (Li et al 2011;Lee et al 2014) and GC (Lee et al 2014;Hu et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Curcuminoid’s Content and Fingerprint Analysis for Authentication and Discrimination of Curcuma xanthorrhiza from Curcuma longa by High-Performance Liquid Chromatography-Diode Array Detector

et al. 2015

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An accurate and reliable method for authentication and discrimination of Curcuma xanthorrhiza (CX) from Curcuma longa (CL) by determining the curcuminoid's content and analyzing the HPLC fingerprint combined with discriminant analysis (DA) was developed. By using the proposed method, it was found that CL had higher amount of all curcuminoid compounds compared to CX. Therefore, these two closely related species could be authenticated and discriminated by the amount of curcuminoids present in the samples. Authentication and discrimination of the two species were also achieved by comparing their HPLC fingerprint chromatograms using their typical marker peaks. To be more convincing, an aid from DA was also used. Combination of HPLC fingerprint analysis and DA gave excellent result that the two species were separated clearly, including CX samples adulterated with CL. The developed method was successfully used for quality control of the two plants.

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“…For example, Photoshop 7.0 and Scion image software were chosen to determine the amount of three curcuminoid in Curcuma longa. The techniques were validated and showed to be accurate and reliable method [16][17] .…”

Section: Discussionmentioning

confidence: 99%

Pharmacognostic evaluation and chrysazin quantitation of Xyris indica flowering heads

Khuniad¹,

Thitikornpong²,

Palanuvej³

et al. 2014

Phcog J.

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Objectives: The present study aimed to establish quality specification of Xyris indica L. flowering heads. The pharmacognostic parameters were investigated. Chrysazin contents were analyzed by TLC image analysis using ImageJ software compared to TLC-densitometry. Methods: X. indica flowering heads from 15 different sources in Thailand were collected. Morphological and physicochemical parameters were characterized. Chrysazin was successively extracted and determined by TLC image analysis using ImageJ software and TLC-densitometry. Results: Macroscopic study was illustrated as whole plant drawing. The microscopic study showed fragment of corolla, seeds, pollen grain and staminode. The pharmacognostic parameters revealed that the loss on drying, total ash, acid-insoluble ash and water content should be not more than 6.90, 2.50, 0.41, and 11.12 of % dry weight respectively while water and ethanol-soluble extractive values should be not less than 6.59 and 4.03 of % dry weight respectively. TLC fingerprint revealed clearly chrysazin yellow fluorescent band at 365 nm. Chrysazin quantitation by TLC image analysis and TLC densitometry were developed and validated. Chrysazin content was 0.022 ± 0.001 % dry weight by both methods. There was no statistically significantly difference between these methods. Conclusion: This study provided pharmacognostic specification and chrysazin content of X. indica flowering heads that can be used for basic quality control and standardization of plant material. TLC image analysis using ImageJ software showed reliable and convenient for analysis of chrysazin content in this crude drug.

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Application of Scion image software to the simultaneous determination of curcuminoids in turmeric (Curcuma longa)

Abstract: TLC image analysis using Scion Image is shown to be a reliable method for the simultaneous analysis of the content of each curcuminoid in turmeric.

Cited by 18 publications

References 15 publications

Assessing the Components of the eIF3 Complex and their Phosphorylation Status

Assessing the Components of the eIF3 Complex and their Phosphorylation Status

Curcuminoid’s Content and Fingerprint Analysis for Authentication and Discrimination of Curcuma xanthorrhiza from Curcuma longa by High-Performance Liquid Chromatography-Diode Array Detector

Pharmacognostic evaluation and chrysazin quantitation of Xyris indica flowering heads

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