Extraction of gypsophila L. rhisomes with supercritical fluid systems, obtaining of nanoparticles and investigation of their emulsiter properties

In this study, the bioactive compounds of Gypsophila Arrostii, which naturally occurs endemically in our country, were extracted and their characteristics were examined in detail. It is significant as the first original study in which valuable bioactive compounds in the rhizomes of Gypsophila were obtained using the supercritical fluid extraction method, and their types and quantities were determined. Hydrophilic bioactive components of Gypsophila rhizomes were obtained by the conventional solvent method, while lipophilic bioactive components were extracted by the supercritical fluid extraction method. The rhizomes were first subjected to supercritical fluid extraction, primarily yielding lipophilic extracts. Then, using the solid residue remaining after SFE and crude Gypsophila rhizomes, solvent extractions were performed with water and ethanol mixtures to obtain hydrophilic extracts. The extracts were analyzed for total phenolic content, total flavonoid content, antioxidant capacity, volatile components, fatty acid composition, tocopherol and tocotrienol contents, types and amounts of triglycerides, phenolic compounds, and saponin contents and types. Nanoemulsions were prepared using the extracts obtained from the extractions, encapsulation techniques were tested to preserve the valuable bioactive compounds in the SFE extracts, and successful results were achieved. In extractions performed with the SFE system, an extraction efficiency of 0.56% was achieved, while solvent extractions from the solid residue remaining after the SFE system yielded an extraction efficiency ranging from 4.8% to 42.5%. From the raw soapwort plant, extraction efficiencies ranging from 4.2% to 42.1% were obtained using solvent extractions. The saponin content in extracts obtained from raw soapwort using solvent extraction systems ranged between 64.3% and 66.1%. In extracts obtained from the solid residue remaining after SFE using solvent extraction systems, the saponin content was found to be between 46.6% and 65.1%. The total phenolic content in the extracts was found to be in the range of 12-16 mg GAE/kg extract for extracts obtained by the solvent extraction system, while it was detected in the range of 59-317 mg GAE/kg extract for those obtained by the SFE system. The total flavonoid content in the extracts was detected in the range of 41-59 mg rutin/kg extract in extracts obtained by the solvent extraction system and in the range of 113-204 mg rutin/kg extract in extracts obtained by the SFE system. The antioxidant activities of the extracts were determined as trolox equivalent amounts, with inhibition percentages ranging from 29-45% in extracts obtained by solvent extraction and from 12-59% in extracts obtained by the SFE system. The volatile components were found to be alpha-Pinene, Eucalyptol (1,8-Cineole), beta-Pinene, delta-3-Carene, Isolongifolene, Caryophyllene, and Beta-Tumerone. The extracts were found to be rich in fatty acid types such as Palmitic acid, Oleic acid, and Linoleic acid. The triglyceride types LLLn, OLLn, OLL, and PLL were determined at high levels. Among tocopherols in the extracts, gamma-tocopherol was found in greater amounts, and among tocotrienols, alpha and beta tocotrienols were higher than others. Phenolic compounds detected in the extracts included gallic acid, procathechin, caffeic acid, 4-dihydroxybenzoic acid, vanillin, rutin, apigenin, and salicylic acid. The presence of steroidal and triterpenic saponins in the Gypsophila has been identified by the color change in the samples. According to the saponin type analysis results carried out by GC-MS, steroidal types such as cholestanol, ergostenol, stigma, sitosterol, and triterpenoid types such as oleanane, ursanoic, lupene saponosides were detected. The amounts of Gypsogenin 3-O glucuronide in the extracts were determined using HPLC and LC systems. The saponin content in the extract obtained by solvent extraction was determined as 325 mg Gypsogenin 3-O glucuronide/kg extract by the LC-MS system. Using the HPLC-UV system, the saponin contents in the extracts obtained by solvent extractions from the crude Gypsophila rhizomes and the solid residues remaining after SFE were found to range between 22-43 mg Gypsogenin 3-O glucuronide/kg extract. The saponin amounts in the extracts obtained from SFE were determined to be 34.2 mg, 22.4 mg, and 18.5 mg Gypsogenin 3-O glucuronide/kg extract in SEP1, SEP2, and SEP3, respectively. Oil/water nanoemulsions were formed using Gypsophila rhizome extracts; the mean particle size distribution of the emulsions prepared with extracts obtained by SFE was determined to be in the range of 125-194 nm, with PDI values between 0.25-0.37. According to these results, a homogeneous appearance and a stable structure were observed. Essential oils present in the extracts obtained by supercritical fluid extraction were encapsulated using a chitosan matrix with TPP, tween 80, and saponin. The average particle sizes of the capsules were determined by DLS, and their morphological structures were identified by FESEM + EDS. Nanoparticles obtained using natural emulsifiers, saponins, are considered to be very useful in the preparation and application of new industrial products. Keywords: Gypsophila arrostii, Supercritical fluid extraction, Chromatography, Nanoemulsion, Saponin, Tocopherol, Terpenoid, Triglyceride, Fatty acid composition, DLS, FESEM