2016
DOI: 10.1039/c6ay01026a
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A fast, adaptable piecewise gradient method for high-throughput quantification of leaf carotenoids using RP-HPLC-PDA

Abstract: Lutein and b-carotene are the major carotenoids in plant leaves, playing crucial roles in photosynthesis.These pigments also have important applications in human and animal health; therefore, it is essential to accurately determine their quantities in crops. A high-throughput RP-HPLC technique was developed for quantifying the levels of these carotenoids in leaf tissue using a piecewise gradient system. The analysis time was reduced to only 10 min, including column re-equilibration, through the use of a short,… Show more

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Cited by 2 publications
(3 citation statements)
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“…Leaf chlorophyll was extracted thrice from 50 mg of ground leaf tissue using 2:1:1 hexane:acetone:ethanol and reconstituted in 5:4:1 acetonitrile:dichloromethane:methanol with 0.5% butylated hydroxytoluene (BHT) after evaporation under a stream of nitrogen (Kormendi et al ). Non‐enzymatic antioxidants were extracted from 100 mg leaf tissue for the Trolox equivalent antioxidant capacity (TEAC) assay twice using 1:1 methanol:water and evaporated by vacuum (Li et al ).…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Leaf chlorophyll was extracted thrice from 50 mg of ground leaf tissue using 2:1:1 hexane:acetone:ethanol and reconstituted in 5:4:1 acetonitrile:dichloromethane:methanol with 0.5% butylated hydroxytoluene (BHT) after evaporation under a stream of nitrogen (Kormendi et al ). Non‐enzymatic antioxidants were extracted from 100 mg leaf tissue for the Trolox equivalent antioxidant capacity (TEAC) assay twice using 1:1 methanol:water and evaporated by vacuum (Li et al ).…”
Section: Methodsmentioning
confidence: 99%
“…To determine chlorophyll content, 5 μl of reconstituted chlorophyll/carotenoid extracts was filtered into 2‐ml high‐performance liquid chromatography (HPLC) vials and analyzed by HPLC using an Agilent 1200 Series HPLC, and separated on a Poroshell 120, EC‐C18, 4.6 × 75 mm, 2.7 μm particle size column with a Poroshell 120 Fast guard column (Agilent Technologies, Mississauga, Canada). Chlorophyll HPLC mobile phase gradients were run according to Kormendi et al (). Chlorophylls were monitored at 645 and 663 nm, and concentrations were determined based on peak area ratios (Ni et al ).…”
Section: Methodsmentioning
confidence: 99%
“…Carotenoids extracted in triplicate batches from Arabidopsis seed or 14-d seedling leaves were analysed as described by Kormendi et al [ 46 ] with minor modifications. Briefly, 50 mg frozen seeds or leaf tissues were ground using RETCH-mill (Retsch Gmbh, 42,787 Haan, Germany) and extracted twice using 750 μL of extraction solvent (2:1:1 hexanes:acetone:ethanol).…”
Section: Methodsmentioning
confidence: 99%