Ying Nie,Yuan Luo,Shajie Luo,Xiujun Cao,Guoxin Song,Chunhui Deng
Polystyrene (PS) microsphere is a kind of attractive extracting medium due to its high stability in different matrices and its particle size can be controlled. The attachment of amphiphilic copolymers to the PS microsphere surface can overcome the drawback of PS relevant to its hydrophobic nature and low wettability. In our previous work, the magnetic composite based on PS microsphere (5 µm) and poly (divinylbenzene-co-N-vinylpyrrolidone, DVB-co-NVP) shell was successfully fabricated and applied for the extraction of safrole in cola drinks. However, the large size and ease of sedimentation limited its application in the enrichment of safrole from blood samples. Considering the adjustability of PS microsphere size, we synthesized the porous PS microspheres with the uniform size of 3 µm and then functionalized with Fe 3 O 4 nanoparticles and poly (DVB-co-NVP) layer in this work. Using the proposed material as extraction sorbent, a simple and fast magnetic solid phase extraction (MSPE) method coupled with HPLC was developed for quantification of safrole in the plasma. Under the optimized conditions, the response to safrole was linear in the range of 0.02–12 µg mL −1 , and the limit of detection (LOD) was 0.006 µg mL −1 . Satisfactory recoveries were obtained between 85.67% and 97.74% (spiked at 0.05, 0.2, 2 µg mL −1 ) and the relative standard deviations (RSDs) for the above spiked levels of the analyte were below 3.9% ( n = 6). The adsorbent can be reused for 6 cycles without a significant loss of extraction capability.
Safrole (SFL) is an IARC class 2B carcinogen. To better understand the mechanism involved in SFL toxicity, we explored the potential interactions between SFL metabolites and RNA. Three guanosine adducts (G1-G3), two adenosine adducts (A1-A2), and two cytosine adducts (C1–C2) were detected by LC-MS/MS in mouse liver S9 incubations, cultured mouse primary hepatocytes, and liver tissues of mice after exposure to SFL. These adducts were chemically synthesized, and one of the guanosine adducts was structurally characterized by 1 H-NMR. Studies in vitro and in vivo showed that SFL was oxidized by cytochrome P450 enzymes to the corresponding 1′-hydroxyl metabolite which was further metabolized by sulfotransferases to form allylic sulfate esters. The formed reactive intermediate(s) subsequently reacted with bases of RNA, leading to RNA adduction, which could play a partial role in the toxicities of SFL through the alteration of RNA biochemical properties and interruption of RNA functions.
Liu Yang,Wang Fang,Guo HuiWen,Zhang Dingkun,Zhang Xiaofei,Wu Zhenfeng,Li Huiting,Xian Yang,Yue Pengfei,Yang Ming
Asarum essential oil (AEO) has been shown to have good pharmacological activities for the anti-inflammatory and analgesic effects, but increasing the dose may cause toxicity. Therefore, we studied the toxic and pharmacodynamic components of AEO by molecular distillation (MD). Anti-inflammatory activity was assessed using RAW264.7 cells. Neurotoxicity was assessed in PC12 cells and the overall toxicity of AEO was evaluated in the mouse acute toxicity assay. The results showed that AEO is primarily composed of safrole, methyl eugenol, and 3,5-dimethoxytoluene. After MD, three fractions were obtained and contained different proportions of volatile compounds relative to the original oil. The heavy fraction had high concentrations of safrole and methyl eugenol, while the light fraction contained high concentrations of α-pinene and β- pinene. The original oil and all three fractions exhibited anti-inflammatory effects, but the light fraction demonstrated more excellent anti-inflammatory activity than the other fractions. Asarum virgin oil and MD products are all neurotoxic. The exposure of PC12 cells to high concentrations of AEO resulted in abnormal nuclei, an increased number of apoptotic cells, increased ROS formation, and decreased SOD levels. Moreover, the results of acute toxicity tests in mice revealed that the light fractions were less toxic than virgin oils and other fractions. In summary, the data suggest that the MD technology enables the enrichment and separation of essential oil components and contributes to the selection of safe concentrations of AEO.
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