Simultaneous determination of different endogenetic plant growth regulators in common green seaweeds using dispersive liquid–liquid microextraction method |
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Authors: | Vishal Gupta Manoj Kumar Harshad Brahmbhatt CRK Reddy Abhiram Seth Bhavanath Jha |
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Institution: | aDiscipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India;bAnalytical Sciences, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India;cAquagri Processing Pvt Ltd, 18, Anand Lok, New Delhi 110049, India |
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Abstract: | A simple and rapid HPLC-based method was developed for simultaneous determination of major classes of plant growth regulators (PGRs) in Monostroma and different species of Ulva. The plant growth regulators determined included gibberellic acid (GA3), indole-3-acetic acid (IAA), abscisic acid (ABA), indole-3-butyric acid (IBA), salicylic acid and kinetin riboside (KR) and their respective elution time was 2.75, 3.3, 3.91, 4.95, 5.39 and 6.59 min. The parameters optimized for distinct separation of PGRs were mobile phase (60:40 methanol and 0.6% acetic acid in water), column temperature (35 °C) and flow rate (1 ml/min). This method presented an excellent linearity (0.2–100 μg/ml) with limit of detection (LOD) as 0.2 μg/ml for ABA, 0.5 μg/ml for KR and salicylic acid, and 1 μg/ml for IAA, IBA and GA3. The precision and accuracy of the method was evaluated after inter and intra day analysis in triplicates. The effect of plant matrix was compensated after spiking and the resultant recoveries estimated were in the range of 80–120%. Each PGR thereby detected were further characterized by ESI-MS analysis. The method optimized in this study determined IBA along with IAA for the first time in the seaweed species investigated except Ulva linza where the former was not detected. In all the species studied, ABA level was detected to be the highest while kinetin riboside was the lowest. In comparison to earlier methods of PGR analysis, sample preparation and analysis time were substantially reduced while allowing determination of more classes of PGRs simultaneously. |
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Keywords: | Plant growth regulator Green seaweed High performance liquid chromatography Dispersive liquid&ndash liquid micro-extraction |
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