College of Pharmacy, Southwest Minzu University, Chengdu 610041, Sichuan Province, China.
National Engineering Institute for the Research and Development of Endangered Medicinal Resources in Southwest China, Guangxi Botanical Garden of Medicinal Plants, Guangxi Province, China.
J Ethnopharmacol. 2019 Apr 24;234:76-84. doi: 10.1016/j.jep.2019.01.026. Epub 2019 Jan 27.
Smilacis Glabrae Rhizoma (SGR), known as Tu-fu-ling in the China, Japan and Korea, is an herb that has been used for clearing damp and detoxification in traditional Chinese medicine for many years. The post-harvest drying of SGR has traditionally been done by the sun, but sometimes sulfur fumigation is used instead due to its low cost and high efficiency. Recent reports show that sulfur fumigation can change the chemical constitution of herbal medicines and decrease their biology activity.
This study will investigate the changes to the chemical constitution, acute toxicity and antioxidant potential of SGR that occur after sulfur fumigation. To date, no studies have investigated these aspects simultaneously.
An ultra-performance liquid chromatography fingerprint method was developed for analysing changes to SGR's chemical constitution caused by sulfur fumigation. The chromatography conditions were as follows: all samples were analysed on a Waters Acquity UHPLC HT3 C18 column; the linear gradient elution was conducted with a mobile phase prepared from acetonitrile and water. All calibration curves showed good linear regression (R > 0.9991) within the tested range. The method was validated for precision, accuracy, limit of detection and quantification. Total flavonoids of the raw and sulfur-fumigated samples were also determined by ultraviolet spectrophotometry. The antioxidant properties of the extracts were evaluated using both DPPH and ABTS radical scavenging assays. The acute toxicities of the raw and sulfur-fumigated samples were investigated.
The results demonstrate that the amounts of astilbin, neoastilbin, neoisoastilbin, isoastilbin, resveratrol and total flavonoids were lower in sulfur-fumigated samples than in raw samples. The antioxidant activity of the sulfur-fumigated samples was also significantly lower. Therefore, sulfur fumigation may cause chemical transformation, alter the chemical constitution, and decrease the bioactivity of SGR. Orally-administered doses did not cause mortality or changes in the general behaviour of tested mice. The LD50 was > 5000 mg/kg DW. However, the high-dose S-SGR mice had significant liver damage and high levels of plasma biochemical parameters (ALT, AST, DBIL, TBIL).
The results of the current study suggest that sulfur fumigation can decrease antioxidant activity in vitro; and that orally-administrated S-SGR is unsafe at doses > 3000 mg/kg dried materia medica. Therefore, sulfur-fumigation processing should be forbidden for SGR until its efficacy and safety has been demonstrated. An alternative method of sulfur fumigation for the post-harvest processing of SGR should also be developed.
菝葜(Smilacis Glabrae Rhizoma,在中国、日本和韩国被称为土茯苓)是一种草药,在中国传统医学中,它已被用于清除湿毒和解毒多年。菝葜的收获后干燥传统上是通过太阳进行的,但由于成本低、效率高,有时也会使用硫磺熏蒸。最近的报告表明,硫磺熏蒸会改变草药的化学结构,降低其生物活性。
本研究旨在调查硫磺熏蒸后菝葜的化学成分、急性毒性和抗氧化潜力的变化。迄今为止,尚无研究同时调查这些方面。
建立了一种超高效液相色谱指纹图谱法分析硫磺熏蒸对菝葜化学成分的影响。色谱条件如下:所有样品均在 Waters Acquity UHPLC HT3 C18 柱上分析;采用乙腈和水制备的流动相进行线性梯度洗脱。所有校准曲线在测试范围内均表现出良好的线性回归(R>0.9991)。该方法对精密度、准确度、检测限和定量限进行了验证。还通过紫外分光光度法测定了生品和硫磺熏蒸品的总黄酮含量。采用 DPPH 和 ABTS 自由基清除试验评估提取物的抗氧化性能。对生品和硫磺熏蒸品进行了急性毒性研究。
结果表明,硫磺熏蒸样品中天门冬酰胺、新天门冬酰胺、新异菝葜素、异菝葜素、白藜芦醇和总黄酮的含量低于生品。硫磺熏蒸样品的抗氧化活性也显著降低。因此,硫磺熏蒸可能导致化学转化,改变化学成分,并降低菝葜的生物活性。口服给予的剂量不会导致受试小鼠死亡或一般行为发生变化。LD50>5000mg/kgDW。然而,高剂量 S-SGR 小鼠的肝脏损伤明显,血浆生化参数(ALT、AST、DBIL、TBIL)水平升高。
本研究结果表明,硫磺熏蒸可降低体外抗氧化活性;口服 S-SGR 剂量>3000mg/kg 干药材时不安全。因此,在证明菝葜的功效和安全性之前,应禁止对其进行硫磺熏蒸处理。还应开发替代硫磺熏蒸的菝葜收获后处理方法。