Exploring the effect of processing on the content of five active ingredients in Cistanche tubulosa based on HPLC-QQQ-MS technology. Cistanche tubulosa is a precious Chinese herbal medicine for nourishing the liver and kidneys, nourishing essence and blood, moistening intestines and promoting bowel movements, and delaying aging. It is used to treat kidney deficiency, female infertility, constipation, improve memory, treat Alzheimer’s disease, and enhance immunity. The components of Cistanche deserticola are diverse, including phenylethanolic glycosides, benzyl alcohol glycosides, cyclohexene ether terpenes and their glycosides, lignans and their glycosaminoglycans, etc. Its extensive pharmacological activity is closely related to the bioactive components it contains. For example, polysaccharides from Cistanche deserticola can effectively scavenge hydroxyl radicals and exhibit significant antioxidant activity. Phenylethanoid glycosides such as verbascoside and echinaceae glycoside can improve chronic high altitude disease by increasing blood oxygen saturation, enhancing body tolerance. Pinecone chrysanthemum glycoside has been proven to have anti Parkinson’s disease effects through research. In order to further explore the active ingredients and pharmacological effects of Cistanche deserticola, we have previously systematically summarized the chemical composition and pharmacological effects of this species.

Due to the obvious and significant pharmacological activity of Cistanche deserticola, there has been relatively more attention paid to its active ingredients, and research on content determination has been carried out in multiple ways. Different research techniques such as tandem coupled reversed-phase hydrophilic interaction chromatography tandem mass spectrometry have been used to determine the composition of different polarities and content ranges in Cistanche deserticola. A study used a relative correction factor calculation method to compare and analyze multiple components of phenylethanoid glycosides in Cistanche deserticola using a marker. UV spectrophotometry, phenol sulfuric acid method, and high-performance liquid chromatography were used to analyze the changes in total phenylethanolic glycosides, total polysaccharides, and four representative components in raw and processed Cistanche deserticola. Some of these research methods cannot be absolutely quantified, while others have low sensitivity. As a high-performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS) technique with low detection limit and high sensitivity, it is more suitable for the analysis of complex and low component content samples of Cistanche deserticola.

Chinese medicine processing is a unique and distinctive pharmaceutical technique in traditional Chinese medicine, which plays an important role in reducing side effects, improving drug efficacy, changing the properties of medicinal materials, and even altering therapeutic effects. Processing can change the key pharmacological substance basis of traditional Chinese medicine, thereby improving its efficacy. Traditional Chinese medicine theory believes that alcohol processing can better enhance the efficacy of kidney tonifying drugs. Therefore, Cistanche deserticola has always been processed with yellow wine and used as medicine. In order to better evaluate the effect of processing on the chemical basis of Cistanche deserticola, we conducted a systematic study on the blood components and metabolomics of Cistanche deserticola before and after processing. Based on the identification and screening of quality markers of compounds in Cistanche deserticola before and after processing, we selected five representative compounds from Cistanche deserticola, including Cistanche glycosides F, verbascoside, isoergosterol glycosides, 2 ‘- acetyl verbascoside, and echinacoside, and analyzed them quantitatively. A method for determining the chemical content of Cistanche deserticola was established on an HPLCQQ-MS instrument. Among the five components, Cistanche deserticola glycoside F belongs to the class of phenylpropanoid oligosaccharides, while the other four are phenylethanoid glycosides, all of which have significant pharmacological activity. Maorui flower glycoside and pineal chrysanthemum glycoside are also used as quality control reference standards in the Chinese Pharmacopoeia. This experiment will provide a theoretical basis for the subsequent processing efficiency and quality evaluation of Cistanche deserticola from a quantitative analysis perspective.

 

The HPLC-QQQ-MS method established in this article is mainly used for the determination of five chemical components in Cistanche deserticola before and after processing. Guided by the quality standard theory of traditional Chinese medicine, the experiment screened the components for content determination based on modern pharmacological research, and ultimately selected Cistanche deserticola glycoside F, verbascoside, isoergosterol glycoside, 2 ‘- acetyl verbascoside, and pineal chrysanthemum glycoside as the indicator components. The experiment followed the extraction method under the content determination item of Cistanche deserticola in the 2020 edition of the Chinese Pharmacopoeia, and extracted samples from both raw and processed Cistanche deserticola products. The liquid phase and mass spectrometry conditions for five chemical components were optimized. The high-performance liquid chromatography tandem triple quadrupole mass spectrometer used in the experiment has the advantages of high resolution, high sensitivity, and rapid analysis. Capable of efficient separation and screening of compounds, quantitative analysis of different molecules, detection of extremely small molecules, and efficient and rapid analysis of complex mixed chemical components. Therefore, the research method established on this platform is very suitable for the detection of components in Cistanche deserticola with slight changes in content before and after processing in this experiment. Based on the results of methodological investigation, it can be found that the method established in this section has high precision, accuracy, and durability. Therefore, this method can be used for the determination of compound content in Cistanche deserticola before and after processing, and its results can be used for quality evaluation of Cistanche deserticola before and after processing.
During the processing of Cistanche deserticola, some compounds may undergo hydrolysis, oxidation, enzymatic, and isomerization reactions due to the influence of heating, resulting in a decrease or increase in their component content. If verbascoside is isomerized to produce isoergosterol glycoside, the content of verbascoside decreases while the content of isoergosterol glycoside increases. However, during the processing, pineal glycoside is prone to hydrolysis reaction, leading to a decrease in its content. Therefore, in order to better exert the efficacy of traditional Chinese medicine, attention should be paid to the changes in the content of the main components during the processing to avoid the phenomenon of insufficient or excessive processing. Overall, the research results of this study provide a theoretical basis for the quality evaluation of Cistanche deserticola in the future, and demonstrate the significance of processing Cistanche deserticola from a quantitative analysis perspective.