Regularity of Toxic Alkaloids during the Combination ofVeratrum nigrumandSalvia miltiorrhizaby UPLC-Q-TOF/MS

Abstract

In traditional chinese medicine, the combination of veratrum nigrum and salvia miltiorrhiza is traditional incom- patibility. In order to investigate the intrinsic relevance of acute toxicity experiment and main toxic alkaloids variation in different ratio, ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to reveal the chemical profile of decoctions of veratrum nigrum and salvia miltiorrhiza in different ratio (10∶0, 10∶1, 5∶1, 2∶1, 1∶1, 1∶2, 1∶5, 1∶10). The analyses were performed on a reversed-phase C18 column using a binary eluent under gradient conditions. At the same time, the trend of toxicity diversity between the veratrum nigrum and salvia miltiorrhiza was explored through acute toxicity experiment of mice. Seventeen alkaloids of veratrum nigrum can be de- tected, The tR-m/z pairs and mean mass can be obtained by selected ion chromatogram, after extracted by Markerlynx XS, the dataset were subjected to principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between samples, and variant changes of veratrum nigrum were observed. After fixed the dosage of veratrum nigrum as LD50, when the ratio of veratrum nigrum was greater than salvia miltiorrhiza, the dissolution of some alkaloids was greater than or equal to the single decoction group of veratrum nigrum, such as verdine, pseudojervine, zygacine, rubijervine, neogermidine, germidine, 3-angeloylzygadenine and so on, with the increase proportion of salvia miltiorrhiza, these alkaloids dissolution showed a declining trend. When the ratio of veratrum nigrum was less than salvia miltiorrhiza, the dissolution of the alkaloids was less than the single decoction group of veratrum nigrum. The result of acute toxicity experiment is matched with the trend of these alkaloids, which showed us these alkaloids may be the main chemical markers of enhanced toxicity during the combination of veratrum nigrum and salvia miltiorrhiza in different ratio. Keywords eighteen incompatible pairs; salvia miltiorrhiza; veratrum nigrum; acute toxicity experiment; ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS)