Seaweed is a macroalgae that is rich in compounds beneficial to health, such as the pigment chlorophyll a. Species Kappaphycus alvarezii is a group of Rhodophyceae which contains a lot of chlorophyll a. Research on seaweed tissue culture on K. alvarezii species in vitro over the last few decades has provided many new discoveries in seaweed cultivation activities. One aspect that will be studied through this research is how to characterize the chlorophyll a content of propagule K alvarezii extract that resulted by tissue culture. This study aimed to characterize and identify the chlorophyll-a content in K. alvarezii propagules resulting from tissue culture to optimize the utilization of seaweed as a raw material for functional food. This research was conducted at the Balai Perikanan Budidaya Laut Lombok (BPBLL) Sekotong, NTB, Indonesia and Laboratorium Kimia Organik Universitas Islam Negeri (UIN) Maulana Malik Ibrahim Malang, Indonesia. This research used experimental methods to characterize the content of chlorophyll a propagule extract of K. alvarezii from tissue culture including the value of water content, extract yield value, isolation results (chromatography, spectrophotometry and FTIR) and identification with LCMS. The results of the identification of chlorophyll a by TLC on isolates of K. alvarezzi obtained blue green spots which have a Rf value of 0.55. The spectral pattern has a maximum absorption of blue waves at a wavelength of 430nm and a maximum absorption of red waves at a wavelength of 662nm with methanol as solvent. In the LCMS method, it was shown that the molecular weight of chlorophyll a was 893.98m/ z.