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Abstract
Aim: Organic substances in leaves of several southwest Australian native species interfere with sensitive colorimetric assays and prevent quantification of inorganic phosphate concentration ([Pi]). We aimed to develop a reproducible routine procedure for treating leaf extracts with activated charcoal (AC) to remove interfering substances, allowing the determination of [Pi] by the malachite green spectrophotometric assay. Methods: Leaf extracts of native plants from southwest Australia in 1% (v/v) acetic acid were treated with 10 mg mL−1 acid-washed AC for removal of interfering substances. Standard solutions (0 to 18 μM Pi) with and without AC treatment were compared to quantify Pi loss. A spiking and recovery test was performed to validate the AC treatment. Results: Leaf extracts treated with AC exhibited distinguishable absorbance peaks for the malachite green-orthophosphate complex between 630 and 650 nm, as opposed to untreated samples. The Pi-adsorption by AC represented a relatively larger fraction of [Pi] in solutions at 0–4 μM Pi range and stabilised at higher [Pi] when maximum adsorption capacity of AC reached at 11.7 μg Pi g−1AC. The Pi recovery after AC treatment in spiked samples ranged between 100 and 111%. Conclusion: The AC treatment successfully removed interfering substances from samples but caused Pi loss. Thus, quantification of [Pi] in AC-treated extracts requires sample [Pi] ≥ 6 μM Pi and the use of AC-treated standards. The error of the AC treatment was minor compared with environmental variability of leaf [Pi]. The AC treatment was a reproducible time- and cost-effective method to remove interfering substances from leaf extracts.
Original language | English |
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Pages (from-to) | 755-764 |
Number of pages | 10 |
Journal | Plant and Soil |
Volume | 476 |
Issue number | 1-2 |
Early online date | 2021 |
DOIs | |
Publication status | Published - Jul 2022 |
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Dive into the research topics of 'Using activated charcoal to remove substances interfering with the colorimetric assay of inorganic phosphate in plant extracts'. Together they form a unique fingerprint.Projects
- 2 Finished
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Facilitation of high leaf phosphorus-use efficiency by nitrate restraint
Lambers, H. (Investigator 01), Finnegan, P. (Investigator 02) & Dassanayake, M. (Investigator 03)
ARC Australian Research Council
1/07/20 → 1/07/24
Project: Research
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Phosphorus-efficient Australian plants: applications for crop improvement
Ranathunge, K. (Investigator 01)
ARC Australian Research Council
1/10/17 → 30/09/22
Project: Research