Amara Amro Abd Al Fattah, El-Masry Mohamed Hesham, Salem Gamal Ali, Baghdadi Hoda Hassan
Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications (SRTA-City), Alexandria P.O. Box 21934, Egypt.
Biotechnology Department, Graduate Studies and Research Institute, Alexandria University, Alexandria P.O. Box 21526, Egypt.
Gels. 2024 Jun 29;10(7):432. doi: 10.3390/gels10070432.
Several cottonseed varieties are cultivated in different countries. Each variety produces a different amount of gossypol as a natural toxic compound. The rising interest in cottonseed products (oil and feed) increases the demand for establishing simple methods for gossypol detection. Silica gel-based methods are ideal for its isolation, purification, and characterization. Silica gel-based methods are variants and can be used as simple methods for tracking plants' compounds. In this study, gossypol was isolated, characterized, and purified as gossypol acetic acid in the form of yellow crystals. Methods used for its characterization were TLC, preparative TLC, silica gel column, UV/IR spectrophotometer, and HPLC (robust spherical silica gel). A comparative study between its amount in both the Egyptian and Chinese varieties was performed. Under the experimental conditions, the Egyptian's cottonseed contains 8.705 gm/kg, while the Chinese's cottonseed contains 5.395 gm/kg. The TLC used in this study proved to be fast, accurate, and inexpensive. It can be used for gossypol acetic acid evaluation and quantification. Additionally, using TLC as a pre-purification step will give a pre-judgment for the sample's purity and quality. This step will protect the expensive HPLC silica gel-based column from any unexpected impurities. During each step, the silica gel itself could be simply removed by paper filtration. Collectively, the different silica gel-based methods as well as the other used methods are recommended for better Gossypol acetic acid isolation, purification, and characterization, as well as for maintaining HPLC columns.
不同国家种植了几种棉花品种。每个品种作为天然有毒化合物产生的棉酚量不同。对棉籽产品（油和饲料）兴趣的增加，使得建立简单的棉酚检测方法的需求也随之增加。基于硅胶的方法非常适合其分离、纯化和表征。基于硅胶的方法有多种变体，可作为追踪植物化合物的简单方法。在本研究中，棉酚被分离、表征并纯化成为黄色晶体形式的棉酚乙酸。用于其表征的方法有薄层色谱法（TLC）、制备型薄层色谱法、硅胶柱法、紫外/红外分光光度计法和高效液相色谱法（坚固球形硅胶）。对埃及和中国品种中的棉酚含量进行了比较研究。在实验条件下，埃及棉籽含棉酚8.705克/千克，而中国棉籽含棉酚5.395克/千克。本研究中使用的薄层色谱法被证明快速、准确且成本低廉。它可用于棉酚乙酸的评估和定量。此外，将薄层色谱法用作预纯化步骤将对样品的纯度和质量给出预判。这一步骤将保护昂贵的基于硅胶的高效液相色谱柱免受任何意外杂质的影响。在每个步骤中，硅胶本身可通过滤纸过滤简单去除。总体而言，推荐使用不同的基于硅胶的方法以及其他使用的方法，以更好地实现棉酚乙酸的分离、纯化和表征，以及维护高效液相色谱柱。
