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Evaluation of a high throughput starch analysis optimised for wood.

Bellasio C, Fini A, Ferrini F - PLoS ONE (2014)

Bottom Line: In this study, a standard method for starch analysis used for food and feed (AOAC standard method 996.11) was optimised to improve precision and accuracy for the analysis of starch in wood.The optimised protocol proved to be remarkably precise and accurate (3%), suitable for a high throughput routine analysis (35 samples a day) of specimens with a starch content between 40 mg and 21 µg.Samples may include lignified organs of coniferous and flowering plants and non-lignified organs, such as leaves, fruits and rhizomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Plant Sciences, University of Sheffield, Sheffield, United Kingdom ; Department of Plant Sciences, University of Cambridge, Cambridge, United Kingdom.

ABSTRACT
Starch is the most important long-term reserve in trees, and the analysis of starch is therefore useful source of physiological information. Currently published protocols for wood starch analysis impose several limitations, such as long procedures and a neutralization step. The high-throughput standard protocols for starch analysis in food and feed represent a valuable alternative. However, they have not been optimised or tested with woody samples. These have particular chemical and structural characteristics, including the presence of interfering secondary metabolites, low reactivity of starch, and low starch content. In this study, a standard method for starch analysis used for food and feed (AOAC standard method 996.11) was optimised to improve precision and accuracy for the analysis of starch in wood. Key modifications were introduced in the digestion conditions and in the glucose assay. The optimised protocol was then evaluated through 430 starch analyses of standards at known starch content, matrix polysaccharides, and wood collected from three organs (roots, twigs, mature wood) of four species (coniferous and flowering plants). The optimised protocol proved to be remarkably precise and accurate (3%), suitable for a high throughput routine analysis (35 samples a day) of specimens with a starch content between 40 mg and 21 µg. Samples may include lignified organs of coniferous and flowering plants and non-lignified organs, such as leaves, fruits and rhizomes.

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Calibration curve for glucose determination.
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pone-0086645-g004: Calibration curve for glucose determination.

Mentions: The glucose assay response was linear between (0 and 40) µg of glucose (Figure 4), corresponding to a concentration of (0 to 13.3) µg ml−1 in the cuvette and to a concentration of (0 to 67) µg ml−1 in the volume occupied by sample + water (0.6 ml, see Table 1). Similarly to the overall starch determination, we evaluated the precision of the glucose analysis by analysing the glucose standard in triplicate for 8 days (Table 6). The C.V. for the glucose assay averaged 0.8%, while the day effect was highly significant. To compensate for this effect, fresh glucose standards were prepared every day (as described in Box 2), instead of using a single calibration curve (Figure 4).


Evaluation of a high throughput starch analysis optimised for wood.

Bellasio C, Fini A, Ferrini F - PLoS ONE (2014)

Calibration curve for glucose determination.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921133&req=5

pone-0086645-g004: Calibration curve for glucose determination.
Mentions: The glucose assay response was linear between (0 and 40) µg of glucose (Figure 4), corresponding to a concentration of (0 to 13.3) µg ml−1 in the cuvette and to a concentration of (0 to 67) µg ml−1 in the volume occupied by sample + water (0.6 ml, see Table 1). Similarly to the overall starch determination, we evaluated the precision of the glucose analysis by analysing the glucose standard in triplicate for 8 days (Table 6). The C.V. for the glucose assay averaged 0.8%, while the day effect was highly significant. To compensate for this effect, fresh glucose standards were prepared every day (as described in Box 2), instead of using a single calibration curve (Figure 4).

Bottom Line: In this study, a standard method for starch analysis used for food and feed (AOAC standard method 996.11) was optimised to improve precision and accuracy for the analysis of starch in wood.The optimised protocol proved to be remarkably precise and accurate (3%), suitable for a high throughput routine analysis (35 samples a day) of specimens with a starch content between 40 mg and 21 µg.Samples may include lignified organs of coniferous and flowering plants and non-lignified organs, such as leaves, fruits and rhizomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Plant Sciences, University of Sheffield, Sheffield, United Kingdom ; Department of Plant Sciences, University of Cambridge, Cambridge, United Kingdom.

ABSTRACT
Starch is the most important long-term reserve in trees, and the analysis of starch is therefore useful source of physiological information. Currently published protocols for wood starch analysis impose several limitations, such as long procedures and a neutralization step. The high-throughput standard protocols for starch analysis in food and feed represent a valuable alternative. However, they have not been optimised or tested with woody samples. These have particular chemical and structural characteristics, including the presence of interfering secondary metabolites, low reactivity of starch, and low starch content. In this study, a standard method for starch analysis used for food and feed (AOAC standard method 996.11) was optimised to improve precision and accuracy for the analysis of starch in wood. Key modifications were introduced in the digestion conditions and in the glucose assay. The optimised protocol was then evaluated through 430 starch analyses of standards at known starch content, matrix polysaccharides, and wood collected from three organs (roots, twigs, mature wood) of four species (coniferous and flowering plants). The optimised protocol proved to be remarkably precise and accurate (3%), suitable for a high throughput routine analysis (35 samples a day) of specimens with a starch content between 40 mg and 21 µg. Samples may include lignified organs of coniferous and flowering plants and non-lignified organs, such as leaves, fruits and rhizomes.

Show MeSH
Related in: MedlinePlus