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H NMR-based metabolomics combined with HPLC-PDA-MS-SPE-NMR for investigation of standardized Ginkgo biloba preparations.

Agnolet S, Jaroszewski JW, Verpoorte R, Staerk D - Metabolomics (2010)

Bottom Line: The standardized extracts originated from Denmark, Italy, Sweden, and United Kingdom, and the results show that (1)H NMR spectra allow simultaneous assessment of the content as well as identity of flavonoid glycosides and TTLs based on a very simple sample-preparation procedure consisting of extraction, evaporation and reconstitution in acetone-d(6).The present study shows that (1)H NMR-based metabolomics is an attractive method for non-selective and comprehensive analysis of Ginkgo extracts.ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-009-0195-x) contains supplementary material, which is available to authorized users.

View Article: PubMed Central - PubMed

ABSTRACT
Commercial preparations of Ginkgo biloba are very complex mixtures prepared from raw leaf extracts by a series of extraction and prepurification steps. The pharmacological activity is attributed to a number of flavonoid glycosides and unique terpene trilactones (TTLs), with largely uncharacterized pharmacological profiles on targets involved in neurological disorders. It is therefore important to complement existing targeted analytical methods for analysis of Ginkgo biloba preparations with alternative technology platforms for their comprehensive and global characterization. In this work, (1)H NMR-based metabolomics and hyphenation of high-performance liquid chromatography, photo-diode array detection, mass spectrometry, solid-phase extraction, and nuclear magnetic resonance spectroscopy (HPLC-PDA-MS-SPE-NMR) were used for investigation of 16 commercially available preparations of Ginkgo biloba. The standardized extracts originated from Denmark, Italy, Sweden, and United Kingdom, and the results show that (1)H NMR spectra allow simultaneous assessment of the content as well as identity of flavonoid glycosides and TTLs based on a very simple sample-preparation procedure consisting of extraction, evaporation and reconstitution in acetone-d(6). Unexpected or unwanted extract constituents were also easily identified in the (1)H NMR spectra, which contrasts traditional methods that depend on UV absorption or MS ionizability and usually require availability of reference standards. Automated integration of (1)H NMR spectral segments (buckets or bins of 0.02 ppm width) provides relative distribution plots of TTLs based on their H-12 resonances. The present study shows that (1)H NMR-based metabolomics is an attractive method for non-selective and comprehensive analysis of Ginkgo extracts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-009-0195-x) contains supplementary material, which is available to authorized users.

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Related in: MedlinePlus

Score plots of four-component PCA model of the region δ 0.5–9.0 of the 1H NMR spectral data of preparation 1–10 and 12–16. a PC1 vs. PC2. b PC2 vs. PC3 [preparation 11 excluded and δ 1.28–1.40, δ 2.04–2.12, and δ 3.28–3.36 regions excluded, 0.01 ppm buckets, data mean-centered]
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Fig3: Score plots of four-component PCA model of the region δ 0.5–9.0 of the 1H NMR spectral data of preparation 1–10 and 12–16. a PC1 vs. PC2. b PC2 vs. PC3 [preparation 11 excluded and δ 1.28–1.40, δ 2.04–2.12, and δ 3.28–3.36 regions excluded, 0.01 ppm buckets, data mean-centered]

Mentions: Preparation 11 was subsequently excluded from the dataset, and so were regions corresponding to residual acetone-d6, acetone, methanol, and the methylene envelope. A new PCA model with four components accounting for 86.3% of the variance was constructed using the rest of the preparations without normalization but with mean-centering. The resulting score plots of PC1 vs. PC2 and PC2 vs. PC3 are shown in Fig. 3a and b, respectively, and the corresponding loading plots in Supplementary Fig. 3. Although a few of the samples displays a large variance between the three replicates, supposedly caused by matrix effects, the inter-group variance is more significant than the intra-group variance. This was shown using an ANOVA test at 95% probability level, and high F-values were confined to spectral regions corresponding to constituents that in the remainder of the work display the highest discriminant power. Other scaling methods like univariate scaling and Pareto scaling were also attempted, but they all resulted in increased intra-group variance without any change in inter-group variance.Fig. 3


H NMR-based metabolomics combined with HPLC-PDA-MS-SPE-NMR for investigation of standardized Ginkgo biloba preparations.

Agnolet S, Jaroszewski JW, Verpoorte R, Staerk D - Metabolomics (2010)

Score plots of four-component PCA model of the region δ 0.5–9.0 of the 1H NMR spectral data of preparation 1–10 and 12–16. a PC1 vs. PC2. b PC2 vs. PC3 [preparation 11 excluded and δ 1.28–1.40, δ 2.04–2.12, and δ 3.28–3.36 regions excluded, 0.01 ppm buckets, data mean-centered]
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2874492&req=5

Fig3: Score plots of four-component PCA model of the region δ 0.5–9.0 of the 1H NMR spectral data of preparation 1–10 and 12–16. a PC1 vs. PC2. b PC2 vs. PC3 [preparation 11 excluded and δ 1.28–1.40, δ 2.04–2.12, and δ 3.28–3.36 regions excluded, 0.01 ppm buckets, data mean-centered]
Mentions: Preparation 11 was subsequently excluded from the dataset, and so were regions corresponding to residual acetone-d6, acetone, methanol, and the methylene envelope. A new PCA model with four components accounting for 86.3% of the variance was constructed using the rest of the preparations without normalization but with mean-centering. The resulting score plots of PC1 vs. PC2 and PC2 vs. PC3 are shown in Fig. 3a and b, respectively, and the corresponding loading plots in Supplementary Fig. 3. Although a few of the samples displays a large variance between the three replicates, supposedly caused by matrix effects, the inter-group variance is more significant than the intra-group variance. This was shown using an ANOVA test at 95% probability level, and high F-values were confined to spectral regions corresponding to constituents that in the remainder of the work display the highest discriminant power. Other scaling methods like univariate scaling and Pareto scaling were also attempted, but they all resulted in increased intra-group variance without any change in inter-group variance.Fig. 3

Bottom Line: The standardized extracts originated from Denmark, Italy, Sweden, and United Kingdom, and the results show that (1)H NMR spectra allow simultaneous assessment of the content as well as identity of flavonoid glycosides and TTLs based on a very simple sample-preparation procedure consisting of extraction, evaporation and reconstitution in acetone-d(6).The present study shows that (1)H NMR-based metabolomics is an attractive method for non-selective and comprehensive analysis of Ginkgo extracts.ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-009-0195-x) contains supplementary material, which is available to authorized users.

View Article: PubMed Central - PubMed

ABSTRACT
Commercial preparations of Ginkgo biloba are very complex mixtures prepared from raw leaf extracts by a series of extraction and prepurification steps. The pharmacological activity is attributed to a number of flavonoid glycosides and unique terpene trilactones (TTLs), with largely uncharacterized pharmacological profiles on targets involved in neurological disorders. It is therefore important to complement existing targeted analytical methods for analysis of Ginkgo biloba preparations with alternative technology platforms for their comprehensive and global characterization. In this work, (1)H NMR-based metabolomics and hyphenation of high-performance liquid chromatography, photo-diode array detection, mass spectrometry, solid-phase extraction, and nuclear magnetic resonance spectroscopy (HPLC-PDA-MS-SPE-NMR) were used for investigation of 16 commercially available preparations of Ginkgo biloba. The standardized extracts originated from Denmark, Italy, Sweden, and United Kingdom, and the results show that (1)H NMR spectra allow simultaneous assessment of the content as well as identity of flavonoid glycosides and TTLs based on a very simple sample-preparation procedure consisting of extraction, evaporation and reconstitution in acetone-d(6). Unexpected or unwanted extract constituents were also easily identified in the (1)H NMR spectra, which contrasts traditional methods that depend on UV absorption or MS ionizability and usually require availability of reference standards. Automated integration of (1)H NMR spectral segments (buckets or bins of 0.02 ppm width) provides relative distribution plots of TTLs based on their H-12 resonances. The present study shows that (1)H NMR-based metabolomics is an attractive method for non-selective and comprehensive analysis of Ginkgo extracts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-009-0195-x) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus