## 5 Ficin Use as an Exfoliant Agent in Cosmetic Applications
### Methods of Extraction and Purification of Ficin  
Ficin extraction and purification typically start with latex collection of a complex mix of ficin and other metabolites [74], which is usually collected from immature green fruits or via incisions on the plant [69; 75]. Following latex collection, several methods are employed to isolate, extract, and purify ficin, each offering distinct advantages. The extraction and purification procedure involves various previous steps, similarly to bromelain, including pre-treatment (cleaning, peeling, and size reduction), cell disruption, and debris removal, including filtration. Conventional ficin extraction begins with latex collection, as mentioned, and the latex is then diluted in a buffer solution in order to prevent enzyme inactivation. Mechanical homogenization, sonication, or enzyme-assisted extraction (EAE) may be employed to enhance yield and preserve enzymatic activity. The crude extract undergoes ultrafiltration and centrifugation to remove insoluble debris and determine the ficin concentration. Precipitation methods, using ammonium sulfate and t-butanol, separate ficin from other proteins, and then dialysis is performed to remove salts and lyophilization or spray drying in order to produce a stable powder [44; 69; 76; 77]. MAE, EAE, and UAE are believed to provide high enzyme recovery, as well (Figure 4) [78; 79; 80]. Three-phase partitioning (TPP) is an efficacious method using ammonium sulfate and tert-butanol to separate ficin from other substances in the crude extract. In TPP, ficin is selectively partitioned into the interfacial precipitate, while contaminants like pigments and lipids move into other phases. TPP is favored for its simplicity, cost-effectiveness, and scalability, and also for providing a high recovery rate of ficin while preventing autolysis [69]. Other less commonly employed techniques, like ATPS [81], RME [82], and supercritical fluid extraction (SFE) [83], have also been confirmed as efficient for ficin extraction, but more recent trials need to be conducted in this context (Figure 4). Chromatographic techniques are also widely used for ficin extraction and purification, and separate proteins based on their physical and chemical properties. Gel filtration-associated chromatography separates proteins according to their specific size, whereas ion exchange chromatography distinguishes them by charge. One example of an effective ion exchange technique is sulphopropyl (SP)-Sepharose [76]. Affinity chromatography is yet another method using specific binding ligands to isolate ficin with high yields [84; 85]. Chromatographic methods may achieve high purity, but are often more expensive and time-consuming than TPP; hence, they are a less preferred option in industry (Figure 4) [68]. Precipitation methods, such as using ammonium sulfate and ethanol or t-butanol, are also employed to selectively precipitate ficin from a crude extract [70]. Ultrafiltration, which  
Figure 4: Schematic representation of ficin extraction and purification. (Parts of this figure were obtained from the free database [[https://smart.servier.com](https://smart.servier.com)]([https://smart.servier.com](https://smart.servier.com)) (accessed on 29 January 2025)). uses membranes with specific molecular weight cut-offs, can concentrate ficin by separating it from smaller molecules and other peptides [86; 87]. Finally, ficin can also be produced from _Ficus carica_ cell cultures, in a more controlled and reproducible way than direct latex extraction. Cell cultures are optimized for enzyme formation and micropropagation techniques, and may be used for large-scale plant production (Figure 4) [88]. The choice of extraction and purification method is dependent on the desired purity, yield, scale, and cost. TPP is particularly useful for its simplicity and cost-effectiveness, while chromatography is valuable for achieving high purity; however, it requires more resources and time.