Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Qixia District, Xianlin Road No. 138, Nanjing 210023, China.
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Qixia District, Xianlin Road No. 138, Nanjing 210023, China.
Phytomedicine. 2022 Jul 20;102:154152. doi: 10.1016/j.phymed.2022.154152. Epub 2022 May 4.
Carbonized traditional Chinese medicine (TCM) is a kind of distinctive traditional medicine, which has been widely used to cure various bleeding syndromes in clinic for over 2000 years. However, there are no effective quality control methods developed on carbonized TCM so far.
This study aimed at developing a processing-associated quality marker (Q-marker) discovery strategy, which would enable to promote the quality control study of carbonized TCM.
Carbonized Typhae Pollen (CTP), a typical carbonized TCM with fantastic efficacy of stanching bleeding and removing blood stasis, was used as an example. First, a ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) method was established to characterize four types of CTP in different processing degrees. Second, chemometric method was applied to screen candidate Q-markers. Third, peak area changes and A changes of each candidate markers in 57 batches samples were described (Traceability and Transitivity). Fourth, systems pharmacology and two high-throughput zebrafish models: cerebral hemorrhage model and thrombus model were used to furtherly screen Q-markers (Effectiveness). Finally, a ultraperformance liquid chromatographic coupled with triple quadrupole tandem mass spectrometry (UPLC-TQ-MS) method was established and applied to quantify Q-markers in additional 10 batches of CTP samples (Measurability).
The chemical profiles of Typhae Pollen during the carbonized process were investigated. Then, 12 candidate compounds were screened in chemometric part. Six Q-markers (isorhamnetin-3-O-neohesperidoside, isorhamnetin-3-O-rutinoside, kaempferol-3-O-neohesperidoside, naringenin, quercetin and isorhamnetin) were subsequently screened out using three principles of Q-markers combined with content changes and two in vivo zebrafish models. Their average contents in additional 10 batches of CTP were 316.8 μg/g, 13.7 μg/g, 6.1 μg/g, 197.8 μg/g, 12.9 μg/g and 199.3 μg/g, respectively. Their content proportion was about 25: 1: 0.5: 15: 1: 15.
A processing-associated Q-marker discovery strategy was developed for carbonized TCM. It might provide a novel insight to solve the problem of 'Chao Tan Cun Xing' in carbonized process.
炭化中药是一种独特的传统药物，在 2000 多年的临床应用中，被广泛用于治疗各种出血症。然而，目前尚未开发出有效的炭化中药质量控制方法。
本研究旨在建立一种与炮制相关的质量标志物（Q-marker）发现策略，以促进炭化中药的质量控制研究。
以炭化蒲黄（CTP）为例，它是一种具有止血化瘀功效的典型炭化中药。首先，采用超高效液相色谱-四极杆飞行时间质谱联用（UPLC-QTOF-MS）方法对不同炮制程度的 4 种 CTP 进行特征分析。其次，应用化学计量学方法筛选候选 Q-marker。然后，描述了 57 批样品中每个候选标志物的峰面积变化和 A 变化（溯源性和传递性）。再次，采用系统药理学和两种高通量斑马鱼模型：脑出血模型和血栓模型，进一步筛选 Q-marker（有效性）。最后，建立超高效液相色谱-三重四极杆串联质谱（UPLC-TQ-MS）方法并应用于另外 10 批 CTP 样品的 Q-marker 定量（可测量性）。
研究了蒲黄在炭化过程中的化学成分变化。然后，在化学计量部分筛选出 12 种候选化合物。最后，结合含量变化和两种体内斑马鱼模型，筛选出 6 种 Q-marker（异鼠李素-3-O-新橙皮苷、异鼠李素-3-O-芦丁、山柰酚-3-O-新橙皮苷、柚皮苷、槲皮素和异鼠李素）。另外 10 批 CTP 样品中，这 6 种 Q-marker 的平均含量分别为 316.8μg/g、13.7μg/g、6.1μg/g、197.8μg/g、12.9μg/g和 199.3μg/g，其含量比例约为 25:1:0.5:15:1:15。
建立了一种与炮制相关的 Q-marker 发现策略，为解决炭化过程中的“超摊陈放”问题提供了新的思路。
