UPLC fingerprint and quantitative analysis of multiple components in rose standard decoction
Rose flowers, also known as rose red, Changchun red, etc., are the dried flowers of Rosa chinensis Jacq. in the Rosaceae family. They are planted in various regions of China and can be harvested throughout the year. They are picked when the flowers are slightly open and dried in the shade or at low temperatures. The medicinal value of rose flowers was first recorded in Li Shizhen’s “Compendium of Materia Medica”. Its taste is sweet, its nature is warm, and it belongs to the liver meridian; It has the effects of promoting blood circulation, regulating menstruation, and soothing the liver to relieve depression. It is mainly used for the treatment of various diseases such as qi stagnation and blood stasis, menstrual disorders, chest and rib distension and pain. Rose flowers contain various chemical components, including flavonoids, phenolic acids, aromatic oils, tannins, and pigments, among which flavonoids and phenolic acids are considered the main medicinal ingredients of rose flowers. Flavonoids are the main components, including quercetin, quercetin, kaempferol-3-O-rhamnoside, kaempferol, succinic acid, gallic acid, ethyl gallate, hyperoside, hesperidin, isoquercitrin, etc. They have biological activities such as antiviral, antioxidant, immune enhancement, and anticancer effects.
The composition of traditional Chinese medicine is complex, and its pharmacological effects are based on the synergistic effects of multiple active ingredients on multiple targets, pathways, and overall systems, exerting regulatory effects on the body. The fingerprint of traditional Chinese medicine is based on the systematic study of the chemical components of traditional Chinese medicine. It comprehensively displays the information of traditional Chinese medicine components from the perspective of substance groups, reflecting the overall situation of traditional Chinese medicine chemical components. It is a comprehensive, quantifiable and effective technical means. This study established fingerprint spectra of 20 batches of rose flower standard decoction using UPLC method combined with chemical pattern recognition technology, analyzed and explored the main characteristic components that affect rose flower standard decoction, and conducted quantitative analysis of multiple indicator components, in order to provide reference for the development of rose flower standard decoction related preparations and quality standard research.

 

To ensure the representativeness of the standard decoction, this study conducted a survey on the information of the main production areas of rose flower raw materials. A total of 20 batches of samples were collected, including on-site samples from major and authentic production areas such as Baoding City in Hebei Province, Zhoukou City in Henan Province, Heze City in Shandong Province, Linyi City in Shandong Province, and Dali Prefecture in Yunnan Province. According to the guiding principles of “Technical Requirements for Quality Control and Standard Development of Traditional Chinese Medicine Formula Granules” and with reference to the “Management Specification for Traditional Chinese Medicine Decoction Room in Medical Institutions”, the pre-treatment methods of standard decoction (decoction times, water addition, decoction time and other indicators and parameters) are investigated to determine the optimal preparation process; Using low-temperature concentration and freeze-drying, a standard rose decoction freeze-dried powder is made.
In the process of establishing the fingerprint method, the effects of extraction solvents (methanol, 70% methanol, 50% methanol, ethanol, 70% ethanol, dilute ethanol), extraction methods (ultrasound, reflux), and extraction times (10, 30, 45, 60 minutes) on the fingerprint were investigated. Considering the adequacy of extraction, stability, and ease of operation, 50% methanol ultrasonic treatment was ultimately chosen for 30 minutes. A diode array detector was used to perform full wavelength scanning (200-400nm) on standard rose soup samples, and the peak numbers, response values, and resolutions of chromatograms at different wavelengths were compared. The results showed that using 254nm as the detection wavelength resulted in a higher number of chromatographic peaks and larger response values, a stable baseline, and less interference. At the same time, the chromatographic conditions of the mobile phase water phase, column temperature, and flow rate were investigated, and it was finally determined to use a 0.2% phosphoric acid solution as the mobile phase water phase, with a flow rate of 0.4 mL/min and column temperature of 35 ℃.
The fingerprint spectrum combined with chemical pattern recognition technology was used to analyze and compare the standard decoction of rose flowers from different origins. The results showed that unsupervised HCA and PCA analysis revealed that samples from different origins could be clustered into four categories. OPLS-DA analysis identified a total of 11 differential biomarkers, mainly flavonoids and phenolic acid components, which played an important role in distinguishing rose standard decoctions from different origins. They can serve as quality control indicators for rose standard decoctions and provide a more comprehensive reference for the raw material sources of rose formula granules.
This study used fingerprint spectra combined with clustering, principal component analysis, orthogonal partial least squares discriminant analysis, and multi index component content determination to conduct a preliminary study on the quality of 20 batches of rose standard decoction. This method can quickly screen key chromatographic peaks that affect the quality of different batches of medicinal materials. Among them, gallic acid, tannic acid, hyperoside, isoquercitrin, astragaloside, quercitrin, and quercetin were identified through high-resolution and reference substance spectral comparison. The results showed that the samples with high content of phenolic acids and flavonoids were collected from Henan Province, belonging to the S11-S13 categories in HCA and PCA analysis. The samples with the lowest content were collected from Yunnan Province, belonging to the first category S1-S6. This can provide a basis for selecting the origin of the raw material collection for the subsequent rose standard decoction. At present, research shows that rose flowers mainly contain flavonoids, flavonoid glycosides, and phenolic acid compounds, which have biological activities such as antioxidant, antiviral, antifungal, anticancer, anti-tumor, and anti-aging. In the later stage, in-depth research will be conducted based on drug efficacy to provide reference for the quality control of rose flower standard decoctions and related preparations.