背景:延龄草。exD.Don是一种富含结构多样的甾体皂苷的民间药草。印度每年对这种草本植物的需求量约为200-500公吨,强调需要进行全面的质量评估。
目的:本研究的目的是开发一种基于超高效液相色谱-蒸发光散射检测器(UHPLC-ELSD)的基于14种特殊代谢产物的质量评估方法。通过网络药理学确定该草药的潜在靶标。
方法:开发了一种UHPLC-ELSD方法,并用14种T.govanianum标记物进行了验证。开发的方法和天然的低共熔溶剂(NADES)辅助提取用于根茎样品(从五个地理上不同的地区收集)中目标特定代谢物的回收率增强研究。此外,对这14种标志物进行了网络药理学方法,以预测T.govanianum可能的生物学靶标.
结果:开发的方法显示出良好的线性(r2:0.940-0.998),检测限(LOD)(2.4-9.0μg),定量限(LOQ)(7.92-29.7μg),精度(日内相对标准偏差[RSD]0.77%-1.96%,日内RSD2.19-4.97%),和准确度(83.24%-118.90%)。NADES样品TG-1*显示了与其乙醇提取物(产量:103.95±5.36mg/g干重)相比的总皂苷含量(TSC)的最高回收率(产量:167.66±4.39mg/g干重)。就TSC而言,与其他分析样品(32.68±1.04-88.22±6.79mg/g)相比,样品TG-1*是最有利的(产量:167.66±4.39mg/g)。五倍子苷D(产量:3.43-28.06mg/g),22β-羟基原薯片(产量:3.22-114.79mg/g),和薯片(产量:1.07-20.82mg/g)被定量为主要代谢物。此外,对靶向的14种标记物的网络药理学分析表明,这些分子可能是治疗神经痛的治疗剂,糖尿病,和痛觉过敏。
结论:当前的研究代表了对从T.govanianum分离的14种化学标记化合物进行同时定量和基于网络药理学分析的第一份报告。
BACKGROUND: Trillium govanianum Wall. ex D.Don is a folk medicinal herb rich in structurally diverse steroidal saponins. The annual demand for this herb in India is about 200-500 metric tons, highlighting the need for a thorough quality assessment.
OBJECTIVE: The objective of this study is to develop an easy and reliable ultrahigh-performance liquid chromatography-evaporative light scattering detector (UHPLC-ELSD)-based quality assessment method with 14 specialised metabolites of T. govanianum and identify the potential targets of this herb using network pharmacology.
METHODS: A UHPLC-ELSD method was developed and validated with 14 markers of T. govanianum. The developed method and natural deep eutectic solvent (NADES)-assisted extraction were utilised for the recovery enhancement study of targeted specialised metabolites from rhizome samples (collected from five geographically distinct areas). In addition, the network pharmacology approach was performed for these 14 markers to predict the plausible biological targets of T. govanianum.
RESULTS: The developed method showed good linearity (r2: 0.940-0.998), limit of detection (LOD) (2.4-9.0 μg), limit of quantification (LOQ) (7.92-29.7 μg), precision (intra-day relative standard deviations [RSDs] 0.77%-1.96% and inter-day RSDs 2.19-4.97%), and accuracy (83.24%-118.90%). NADES sample TG-1* showed the highest recovery (yield: 167.66 ± 4.39 mg/g of dry weight) of total saponin content (TSC) as compared to its hydroethanolic extract (yield: 103.95 ± 5.36 mg/g of dry weight). Sample TG-1* was the most favourable (yield: 167.66 ± 4.39 mg/g) in terms of TSC as compared to other analysed samples (32.68 ± 1.04-88.22 ± 6.79 mg/g). Govanoside D (yield: 3.43-28.06 mg/g), 22β-hydroxyprotodioscin (yield: 3.22-114.79 mg/g), and dioscin (yield: 1.07-20.82 mg/g) were quantified as the major metabolites. Furthermore, network pharmacology analysis of targeted 14 markers indicated that these molecules could be possible therapeutic agents for managing neuralgia, diabetes mellitus, and hyperalgesia.
CONCLUSIONS: The current study represents the first report for the simultaneous quantification and a network pharmacology-based analysis of 14 chemical marker compounds isolated from T. govanianum.