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Assessing temporal flux of plant hormones in stored processing potatoes using high definition accurate mass spectrometry.

Ordaz-Ortiz JJ, Foukaraki S, Terry LA - Hortic Res (2015)

Bottom Line: Sylvana and Russet Burbank stored for up to 30 weeks at 6 °C under ambient air conditions.The method achieved a lowest limit of detection of 0.22 ng g(-1) of dry weight and a limit of quantification of 0.74 ng g(-1) dry weight (zeatin riboside), and was able to recover, detect and quantify a total of 12 plant hormones spiked on flesh and skin of potato tubers.In addition, the mass accuracy for all compounds (<5 ppm) was evaluated.

View Article: PubMed Central - PubMed

Affiliation: Plant Science Laboratory, Cranfield University , Bedfordshire, MK43 0AL, UK.

ABSTRACT
Plant hormones are important molecules which at low concentration can regulate various physiological processes. Mass spectrometry has become a powerful technique for the quantification of multiple classes of plant hormones because of its high sensitivity and selectivity. We developed a new ultrahigh pressure liquid chromatography-full-scan high-definition accurate mass spectrometry method, for simultaneous determination of abscisic acid and four metabolites phaseic acid, dihydrophaseic acid, 7'-hydroxy-abscisic acid and abscisic acid glucose ester, cytokinins zeatin, zeatin riboside, gibberellins (GA1, GA3, GA4 and GA7) and indole-3-acetyl-L-aspartic acid. We measured the amount of plant hormones in the flesh and skin of two processing potato cvs. Sylvana and Russet Burbank stored for up to 30 weeks at 6 °C under ambient air conditions. Herein, we report for the first time that abscisic acid glucose ester seems to accumulate in the skin of potato tubers throughout storage time. The method achieved a lowest limit of detection of 0.22 ng g(-1) of dry weight and a limit of quantification of 0.74 ng g(-1) dry weight (zeatin riboside), and was able to recover, detect and quantify a total of 12 plant hormones spiked on flesh and skin of potato tubers. In addition, the mass accuracy for all compounds (<5 ppm) was evaluated.

No MeSH data available.


Related in: MedlinePlus

(a) UHPLC Q-TOF MS overlaid EICs of a mixture of 14 authentic plant hormone standards, section 1 refers to the first time segment which was acquired in positive ionization mode and section 2 to the second time segment which was acquired in negative ionization mode. (b) Overlaid extracted ion chromatograms of skin tissue extract from potato sample. Compounds are as follows: 0.87, trans-zeatin; 0.98, cis-zeatin; 1.49, trans-zeatin riboside; 1.63, cis-zeatin riboside; 2.06, dihydrophaseic acid; 2.41, indole-3-acetyl-l-aspartic acid, 3.03, gibberellin 3; 3.14, gibberellin 1; 3.39, abscisic acid glucose ester; 3.49, phaseic acid; 4.19, 7′-hydroxy-abscisic acid; 5.50, abscisic acid; 8.77, gibberellin 7; 8.97, gibberellin 4.
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fig1: (a) UHPLC Q-TOF MS overlaid EICs of a mixture of 14 authentic plant hormone standards, section 1 refers to the first time segment which was acquired in positive ionization mode and section 2 to the second time segment which was acquired in negative ionization mode. (b) Overlaid extracted ion chromatograms of skin tissue extract from potato sample. Compounds are as follows: 0.87, trans-zeatin; 0.98, cis-zeatin; 1.49, trans-zeatin riboside; 1.63, cis-zeatin riboside; 2.06, dihydrophaseic acid; 2.41, indole-3-acetyl-l-aspartic acid, 3.03, gibberellin 3; 3.14, gibberellin 1; 3.39, abscisic acid glucose ester; 3.49, phaseic acid; 4.19, 7′-hydroxy-abscisic acid; 5.50, abscisic acid; 8.77, gibberellin 7; 8.97, gibberellin 4.

Mentions: The first step of the analysis for all 12 PHs was to record full-scan spectral data chromatograms. The exact mass to charge ratio (m/z) of the molecules was extracted from the raw data or total ion chromatogram generating an EIC (Figure 1); and the retention time and accurate mass of each compound (standard and internal standard) were obtained. The chromatograms shown on Figure 1 are both EICs, which are free of noise as each peak depicts only the area underneath the accurate mass of each targeted compound where it is most found. The accurate masses of the molecular ions in observed protonated molecule (M+H)+ and deprotonated molecule (M–H)− for positive and negative ionization modes respectively were used for both confirmation and quantification purposes. The chromatograms obtained (EICs) were processed through the quantitative analysis software, which provided the mass accuracy values (difference between measured mass and calculated exact or theoretical mass divided by exact mass and expressed in ppm). Accurate mass measurements were obtained (Table 1) and the errors obtained for most compounds were well below the established accuracy threshold of 5 ppm for unambiguous identification of the 12 PHs and their corresponding deuterated forms. Further unambiguous confirmation of the identity of the PHs was carried out using targeted MS/MS experiments under collision-induced dissociation conditions. The product ion spectra obtained were then compared to match with the standard solution samples (Supplementary Fig. S1) and with data from the published literature obtained with the multiple reaction monitoring method using a triple quadrupole instrument.


Assessing temporal flux of plant hormones in stored processing potatoes using high definition accurate mass spectrometry.

Ordaz-Ortiz JJ, Foukaraki S, Terry LA - Hortic Res (2015)

(a) UHPLC Q-TOF MS overlaid EICs of a mixture of 14 authentic plant hormone standards, section 1 refers to the first time segment which was acquired in positive ionization mode and section 2 to the second time segment which was acquired in negative ionization mode. (b) Overlaid extracted ion chromatograms of skin tissue extract from potato sample. Compounds are as follows: 0.87, trans-zeatin; 0.98, cis-zeatin; 1.49, trans-zeatin riboside; 1.63, cis-zeatin riboside; 2.06, dihydrophaseic acid; 2.41, indole-3-acetyl-l-aspartic acid, 3.03, gibberellin 3; 3.14, gibberellin 1; 3.39, abscisic acid glucose ester; 3.49, phaseic acid; 4.19, 7′-hydroxy-abscisic acid; 5.50, abscisic acid; 8.77, gibberellin 7; 8.97, gibberellin 4.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4595984&req=5

fig1: (a) UHPLC Q-TOF MS overlaid EICs of a mixture of 14 authentic plant hormone standards, section 1 refers to the first time segment which was acquired in positive ionization mode and section 2 to the second time segment which was acquired in negative ionization mode. (b) Overlaid extracted ion chromatograms of skin tissue extract from potato sample. Compounds are as follows: 0.87, trans-zeatin; 0.98, cis-zeatin; 1.49, trans-zeatin riboside; 1.63, cis-zeatin riboside; 2.06, dihydrophaseic acid; 2.41, indole-3-acetyl-l-aspartic acid, 3.03, gibberellin 3; 3.14, gibberellin 1; 3.39, abscisic acid glucose ester; 3.49, phaseic acid; 4.19, 7′-hydroxy-abscisic acid; 5.50, abscisic acid; 8.77, gibberellin 7; 8.97, gibberellin 4.
Mentions: The first step of the analysis for all 12 PHs was to record full-scan spectral data chromatograms. The exact mass to charge ratio (m/z) of the molecules was extracted from the raw data or total ion chromatogram generating an EIC (Figure 1); and the retention time and accurate mass of each compound (standard and internal standard) were obtained. The chromatograms shown on Figure 1 are both EICs, which are free of noise as each peak depicts only the area underneath the accurate mass of each targeted compound where it is most found. The accurate masses of the molecular ions in observed protonated molecule (M+H)+ and deprotonated molecule (M–H)− for positive and negative ionization modes respectively were used for both confirmation and quantification purposes. The chromatograms obtained (EICs) were processed through the quantitative analysis software, which provided the mass accuracy values (difference between measured mass and calculated exact or theoretical mass divided by exact mass and expressed in ppm). Accurate mass measurements were obtained (Table 1) and the errors obtained for most compounds were well below the established accuracy threshold of 5 ppm for unambiguous identification of the 12 PHs and their corresponding deuterated forms. Further unambiguous confirmation of the identity of the PHs was carried out using targeted MS/MS experiments under collision-induced dissociation conditions. The product ion spectra obtained were then compared to match with the standard solution samples (Supplementary Fig. S1) and with data from the published literature obtained with the multiple reaction monitoring method using a triple quadrupole instrument.

Bottom Line: Sylvana and Russet Burbank stored for up to 30 weeks at 6 °C under ambient air conditions.The method achieved a lowest limit of detection of 0.22 ng g(-1) of dry weight and a limit of quantification of 0.74 ng g(-1) dry weight (zeatin riboside), and was able to recover, detect and quantify a total of 12 plant hormones spiked on flesh and skin of potato tubers.In addition, the mass accuracy for all compounds (<5 ppm) was evaluated.

View Article: PubMed Central - PubMed

Affiliation: Plant Science Laboratory, Cranfield University , Bedfordshire, MK43 0AL, UK.

ABSTRACT
Plant hormones are important molecules which at low concentration can regulate various physiological processes. Mass spectrometry has become a powerful technique for the quantification of multiple classes of plant hormones because of its high sensitivity and selectivity. We developed a new ultrahigh pressure liquid chromatography-full-scan high-definition accurate mass spectrometry method, for simultaneous determination of abscisic acid and four metabolites phaseic acid, dihydrophaseic acid, 7'-hydroxy-abscisic acid and abscisic acid glucose ester, cytokinins zeatin, zeatin riboside, gibberellins (GA1, GA3, GA4 and GA7) and indole-3-acetyl-L-aspartic acid. We measured the amount of plant hormones in the flesh and skin of two processing potato cvs. Sylvana and Russet Burbank stored for up to 30 weeks at 6 °C under ambient air conditions. Herein, we report for the first time that abscisic acid glucose ester seems to accumulate in the skin of potato tubers throughout storage time. The method achieved a lowest limit of detection of 0.22 ng g(-1) of dry weight and a limit of quantification of 0.74 ng g(-1) dry weight (zeatin riboside), and was able to recover, detect and quantify a total of 12 plant hormones spiked on flesh and skin of potato tubers. In addition, the mass accuracy for all compounds (<5 ppm) was evaluated.

No MeSH data available.


Related in: MedlinePlus