KM Science Research Division, Korea Institute of Oriental Medicine, Daejeon 34054, Korea.
Advanced Toxicity Evaluation Team 2, Biotoxtech Co., Ltd., Cheongju 28115, Korea.
Molecules. 2022 Jun 24;27(13):4066. doi: 10.3390/molecules27134066.
Siryung-tang (SRT) is a traditional herbal prescription containing Oryeong-san and Soshiho-tang that is used to treat digestive system diseases. We performed safety evaluations of SRT based on genotoxicity and developed an assay for quality control using high-performance liquid chromatography with a photodiode array detector. Genotoxicity was evaluated based on bacterial reverse mutation ( TA1535, TA98, TA100, and TA1537, and WP2 uvrA), chromosomal aberration (Chinese hamster lung cells), and micronucleus (mouse) tests. Quality control analysis was conducted using a SunFire C column and gradient elution with a distilled water-acetonitrile mobile phase system containing 0.1% (/) formic acid for 12 markers (5-(hydroxy-methyl)furfural, 3,4-dihydroxybenzaldehyde, liquiritin apioside, liquiritin, coumarin, baicalin, wogonoside, cinnamaldehyde, baicalein, glycyrrhizin, wogonin, and atractylenolide III). SRT showed no genotoxicity in three tests. Ames tests showed that SRT at 313-5000 μg/plate did not significantly increase the number of revertant colonies with or without metabolic activation among five bacterial strains. Moreover, in vivo micronucleus testing showed that SRT did not increase the frequency of bone marrow micronuclei. The number of chromosomal aberrations associated with SRT was similar to that observed in the negative controls. The 12 markers were detected at 0.04-16.86 mg/g in a freeze-dried SRT sample and completely eluted within 45 min. The extraction recovery was 95.39-104.319% and the relative standard deviation value of the precision was ≤2.09%. Our study will be used as basic data for the safety and standardization of SRT.
实肠汤（Siryung-tang，SRT）是一种传统的草药方剂，包含 Oryeong-san 和 Soshiho-tang，用于治疗消化系统疾病。我们基于遗传毒性对 SRT 进行了安全性评估，并开发了一种使用高效液相色谱-光电二极管阵列检测器的质量控制分析方法。遗传毒性评估基于细菌回复突变（TA1535、TA98、TA100 和 TA1537 以及 WP2 uvrA）、染色体畸变（中国仓鼠肺细胞）和微核（小鼠）试验。质量控制分析使用 SunFire C 柱和梯度洗脱，以蒸馏水-乙腈流动相系统（含 0.1%（/）甲酸）洗脱 12 个标志物（5-（羟甲基）糠醛、3,4-二羟基苯甲醛、甘草酸苷、甘草苷、香豆素、黄芩苷、木犀草苷、桂皮醛、黄芩素、甘草酸、汉黄芩苷和白术内酯 III）。SRT 在三项测试中均未显示出遗传毒性。Ames 测试表明，SRT 在 313-5000μg/ 平板时，无论是否有代谢激活，在 5 株细菌中均未显著增加回复突变菌落的数量。此外，体内微核试验表明，SRT 未增加骨髓微核的频率。SRT 相关的染色体畸变数量与阴性对照相似。在冻干的 SRT 样品中检测到 12 个标志物，浓度为 0.04-16.86mg/g，且在 45min 内完全洗脱。提取回收率为 95.39-104.319%，精密度的相对标准偏差值≤2.09%。我们的研究将作为 SRT 安全性和标准化的基础数据。
