A rapid and sensitive method for determination of carotenoids in plant tissues by high performance liquid chromatography

Gupta, Prahlad; Sreelakshmi, Yellamaraju; Sharma, Rameshwar

doi:10.1186/s13007-015-0051-0

Cited by 138 publications

(114 citation statements)

References 35 publications

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“…Chlorophylls and carotenoids were extracted from leaves from 7–8 internodes of 45-day-old plants in 80% (v/v) acetone using the protocol of Makeen et al (2007) and their amounts were calculated using the equation of Lichtenthaler (1987). Carotenoids were extracted from the pericarp of MG, BR and RR fruits using the procedure of Gupta et al (2015). To avoid photooxidation, the entire procedure was performed under dim light.…”

Section: Methodsmentioning

confidence: 99%

Nitric Oxide Overproduction in Tomato shr Mutant Shifts Metabolic Profiles and Suppresses Fruit Growth and Ripening

et al. 2016

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Nitric oxide (NO) plays a pivotal role in growth and disease resistance in plants. It also acts as a secondary messenger in signaling pathways for several plant hormones. Despite its clear role in regulating plant development, its role in fruit development is not known. In an earlier study, we described a short root (shr) mutant of tomato, whose phenotype results from hyperaccumulation of NO. The molecular mapping localized shr locus in 2.5 Mb region of chromosome 9. The shr mutant showed sluggish growth, with smaller leaves, flowers and was less fertile than wild type. The shr mutant also showed reduced fruit size and slower ripening of the fruits post-mature green stage to the red ripe stage. Comparison of the metabolite profiles of shr fruits with wild-type fruits during ripening revealed a significant shift in the patterns. In shr fruits intermediates of the tricarboxylic acid (TCA) cycle were differentially regulated than WT indicating NO affected the regulation of TCA cycle. The accumulation of several amino acids, particularly tyrosine, was higher, whereas most fatty acids were downregulated in shr fruits. Among the plant hormones at one or more stages of ripening, ethylene, Indole-3-acetic acid and Indole-3-butyric acid increased in shr, whereas abscisic acid declined. Our analyses indicate that the retardation of fruit growth and ripening in shr mutant likely results from the influence of NO on central carbon metabolism and endogenous phytohormones levels.

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Section: Methodsmentioning

confidence: 99%

Nitric Oxide Overproduction in Tomato shr Mutant Shifts Metabolic Profiles and Suppresses Fruit Growth and Ripening

et al. 2016

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“…This report Gupta 4 Yu 15 Maurer 25 Arnold 29 Wehrens 32 and 109.4% for lutein and b-carotene, respectively. Precision was evaluated using % CV and was less than 9% for unspiked, spiked, and standard samples.…”

Section: Referencementioning

confidence: 68%

“…compared to gradient methods that typically involve 3-13 min of added post-run time (Table 3). 4,15,29 Although both methanol and MTBE are miscible, in our analysis, different chromatograms were produced depending on whether the solvents were premixed (offline) or mixed online via the HPLC pump (Fig. This simple solvent system has been utilized by various gradient methods 4,15,29 (Table 3), where at least 25 min of separation time is required, not including post-conditioning.…”

Section: Evaluation Of Mobile Phase and Solvent Systemmentioning

confidence: 99%

“…1,2 Their catabolic products include the growth regulators strigolactones and abscisic acid as well as volatile compounds, such as b-ionone which has been implicated in plant-insect interactions. 1,2,4 The ability of carotenoids to attenuate oxidative damage to DNA and proteins has provided numerous potential nutritional and health applications. 1,2,4 The ability of carotenoids to attenuate oxidative damage to DNA and proteins has provided numerous potential nutritional and health applications.…”

Section: Introductionmentioning

confidence: 99%

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A fast, adaptable piecewise gradient method for high-throughput quantification of leaf carotenoids using RP-HPLC-PDA

et al. 2016

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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, efficientPoroshell column with an HPLC-PDA instrument. The method was successfully validated, resulting in sub-mg mL À1 quantification limits for both analytes. To demonstrate the adaptability of the method, application to both saponified and unsaponified leaf samples was performed. The piecewise gradient allows for the customizability of the mobile phase selectivity towards specific sample components and can potentially be adapted to screen multiple tissues of plant varieties and mutants with more extensive carotenoid profiles.

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“…However, recently Gupta et al. resolved efficiently 15 CARs and their isomers in shorter runtime of 20 min.…”

Section: Resultsmentioning

confidence: 99%

Optimization and development of a high‐performance liquid chromatography method for the simultaneous determination of vitamin E and carotenoids in tomato fruits

Irakli¹,

Chatzopoulou²,

Kadoglidou³

et al. 2016

J of Separation Science

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A simple and reliable high-performance liquid chromatography method was developed and validated for the simultaneous determination of lipophilic antioxidants in tomato fruits using C30 column operated at 15°C and a gradient mobile phase based on acetonitrile/methanol/dichloromethane in a total run time of 30 min. Diode array and fluorescence detectors were used respectively for the detection of carotenoids (lutein, zeaxanthin, cryptoxanthin, lycopene, and β-carotene) and vitamin E analogs (α-, β-, γ-, and δ-tocopherols, and tocotrienols). The best extraction yield of analytes in tomato fruits was achieved by employing ethyl acetate/hexane (1:1, v/v) after several treatments with various solvents. In addition, low extraction yields were obtained for carotenoids compared to tocopherols by adopting solid-phase extraction as a second clean-up step. The method was validated on the basis of recovery, precision, linearity, and limit of detection and quantification using spiked tomato samples. The method was applied to cherry and medium-sized tomato fruits. Lycopene was found to be present in largest amount in tomato pulp, followed by β-carotene and lutein. Due to its simplicity, rapidity, and efficiency, the method is suitable for routine analysis of lipophilic antioxidants in tomato fruits, and may also be applied to other vegetables of similar phytochemical profiles.

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A rapid and sensitive method for determination of carotenoids in plant tissues by high performance liquid chromatography

Cited by 138 publications

References 35 publications

Nitric Oxide Overproduction in Tomato shr Mutant Shifts Metabolic Profiles and Suppresses Fruit Growth and Ripening

Nitric Oxide Overproduction in Tomato shr Mutant Shifts Metabolic Profiles and Suppresses Fruit Growth and Ripening

A fast, adaptable piecewise gradient method for high-throughput quantification of leaf carotenoids using RP-HPLC-PDA

Optimization and development of a high‐performance liquid chromatography method for the simultaneous determination of vitamin E and carotenoids in tomato fruits

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