Ke Zhong-Cheng, Yang Nan, Hou Xue-Feng, Wang Ai-Dong, Feng Liang, Jia Xiao-Bin
College of Chemistry, Huangshan University, Huangshan 245041, China.
College of Third Clinical Medical, Nanjing University of Chinese Medicine, Nanjing 210028, China.
Zhongguo Zhong Yao Za Zhi. 2016 Oct;41(20):3839-3845. doi: 10.4268/cjcmm20162021.
In order to clarify the effect of intestinal flora on the absorption and metabolism of paeoniflorin in vivo, the metabolism of paeoniflorin by rat intestinal flora was studied under the in vitro anaerobic condition. Paeoniflorin was incubated with rat anaerobic intestinal flora for 48 h, and UPLC was used to detect the changes of paeoniflorin at different incubation time points under the following chromatographic conditions：WelchromTM C₁₈ chromatographic column (4.6 mm×100 mm, 5 μm), with 0.1% formic acid(A)-acetonitrile(B) as the mobile phase for gradient elution. The flow rate was 0.4 mL•min⁻¹, and column temperature was 30 ℃. UPLC-Q-TOF-MS with positive ion mode(ESI ion source) was applied to investigate the structural characterization of metabolic products. The structures of the metabolites were identified by accurate molecular weight, TOF-MS/MS fragmentation information, combined with retention time and literature data review, and the intestinal metabolic rules were then analyzed. After incubation for 24 h, the paeoniflorin was metabolized completely, and the resulting metabolites(albiflorin, albiflorinaglycone, deacylate albiflorin, deacylate albiflorin aglycone and paeonilactone-B) were detected in rat intestinal flora. The metabolic pathway analysis showed that the isolated rat intestinal flora first transformed peoniflorin into albiflorin, and then further metabolized by glucose removal, phenyl group removal, or four-membered ring pyrolysis and rearrangement. Paeoniflorin was gradually transformed into more hydrophobic metabolites with smaller molecular mass, which were better absorbed by the intestinal tract.
为阐明肠道菌群对芍药苷体内吸收和代谢的影响，在体外厌氧条件下研究了大鼠肠道菌群对芍药苷的代谢。将芍药苷与大鼠厌氧肠道菌群孵育48小时，采用超高效液相色谱法（UPLC）在以下色谱条件下检测不同孵育时间点芍药苷的变化：WelchromTM C₁₈色谱柱（4.6 mm×100 mm，5μm），以0.1%甲酸（A）-乙腈（B）为流动相进行梯度洗脱。流速为0.4 mL•min⁻¹，柱温为30℃。采用正离子模式（电喷雾离子源）的超高效液相色谱-四极杆-飞行时间质谱联用仪（UPLC-Q-TOF-MS）研究代谢产物的结构特征。通过精确分子量、TOF-MS/MS碎片信息，并结合保留时间和文献数据回顾来鉴定代谢产物的结构，进而分析肠道代谢规律。孵育24小时后，芍药苷完全代谢，在大鼠肠道菌群中检测到产生的代谢产物（芍药内酯苷、芍药内酯苷元、去酰基芍药内酯苷、去酰基芍药内酯苷元及芍药内酯B）。代谢途径分析表明，分离得到的大鼠肠道菌群首先将芍药苷转化为芍药内酯苷，然后通过去葡萄糖、去苯基或四元环裂解和重排进一步代谢。芍药苷逐渐转化为分子质量更小、疏水性更强的代谢产物，更易被肠道吸收。
