The sap expelled from the fresh harvest of Kappaphycus alvarezii , a red seaweed growing in tropical waters, has been reported to be a potent foliar spray. Tandem mass spectrometry of various organic extracts of the sap confirmed the presence of the plant growth regulators (PGRs) indole 3-acetic acid, gibberellin GA(3), kinetin, and zeatin. These PGRs were quantified in fresh state and after 1 year of storage by ESI-MS without recourse to chromatographic separation. Quantification was validated against HPLC data. The results may be useful in correlating with the efficacy of the sap. The methodology was extended to two other seaweeds. The method developed is convenient and precise and may find application in other agricultural formulations containing these growth hormones.