Guo Qingquan, Wu Shaojuan, Liang Wenyao, Tan Jianhua, Liu Xiangmei, Yuan Yuxi, Li Xiaohong, Zhao Haishan
School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China.
Guangzhou Quality Supervision and Testing Institute, Guangzhou 511447, China.
J Ethnopharmacol. 2022 Apr 6;287:114963. doi: 10.1016/j.jep.2021.114963. Epub 2021 Dec 28.
Glabridin, extracted from Glycyrrhiza glabra L., is widely used for the treatment of hyperpigmentation because of its anti-inflammatory and antioxidant activities and its ability to inhibit melanin synthesis. This led to the strict regulation of its quality and safety. However, traditional quality control methods used for plant extracts cannot reflect the product quality owing to multiple unknown impurities, which necessitates the further analysis of impurities.
The study identified the toxic impurities of glabridin and their toxicological mechanism.
In total, 10 glabridin samples from different sources were quantified using high-performance liquid chromatography. Sample toxicities were evaluated using zebrafish and cell models. To identify impurities, samples with different toxicity were analyzed by ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry. The toxicity of related impurities was verified in the zebrafish model. Phalloidin stain was used to evaluate subtle changes in myofibril alignment.
Although glabridin content in the samples was similar, there were significant differences in toxicity. The results were verified using four different mammalian cell lines. Higher contents of glabrone and glabrol were identified in the sample with the highest toxicity. In the zebrafish model, the addition of glabrol reduced the LC of glabridin to 9.224, 6.229, and 5.370 μM at 48, 72, and 96 h post-fertilization, respectively, whereas glabrone did not have any toxic effect. Phalloidin staining indicated that a glabrol impurity exacerbates the myotoxicity of glabridin in zebrafish embryos.
Glabrol, but not glabrone, was identified as a key impurity that increased glabridin toxicity. This finding indicates that controlling glabrol content is necessary during glabridin product production.
光甘草定是从光果甘草中提取的，因其具有抗炎、抗氧化活性以及抑制黑色素合成的能力，被广泛用于治疗色素沉着。这导致了对其质量和安全性的严格监管。然而，由于存在多种未知杂质，用于植物提取物的传统质量控制方法无法反映产品质量，因此有必要对杂质进行进一步分析。
本研究旨在鉴定光甘草定的有毒杂质及其毒理学机制。
总共使用高效液相色谱法对10个不同来源的光甘草定样品进行了定量分析。使用斑马鱼和细胞模型评估样品的毒性。为了鉴定杂质，采用超高效液相色谱-四极杆-轨道阱质谱联用技术对具有不同毒性的样品进行了分析。在斑马鱼模型中验证了相关杂质的毒性。使用鬼笔环肽染色评估肌原纤维排列的细微变化。
尽管样品中的光甘草定含量相似，但毒性存在显著差异。使用四种不同的哺乳动物细胞系验证了结果。在毒性最高的样品中鉴定出较高含量的光甘草酮和光甘草醇。在斑马鱼模型中，添加光甘草醇后，受精后48、72和96小时时光甘草定的半数致死浓度分别降至9.224、6.229和5.370μM，而光甘草酮没有任何毒性作用。鬼笔环肽染色表明，光甘草醇杂质会加剧光甘草定对斑马鱼胚胎的肌毒性。
光甘草醇而非光甘草酮被鉴定为增加光甘草定毒性的关键杂质。这一发现表明，在光甘草定产品生产过程中控制光甘草醇含量是必要的。
