Quality evaluation of Yupingfeng powder based on HPLC fingerprint and multi-component content determination combined with chemometrics
Yuping Fengsan was first recorded in Zhu Zhenheng’s “Danxi Heart Method” of the Yuan Dynasty. It consists of three herbs: windproof, astragalus, and Atractylodes macrocephala. In traditional Chinese medicine, there is a saying that “Yuping combination is less and more refined, while Qishu windproof and tripod foot movement”. This formula nourishes qi, solidifies the exterior, and stops sweating. It is used for those with superficial deficiency but not firm, self sweating and aversion to wind, pale complexion, or those who are susceptible to wind evil due to body deficiency. Huangqi, a traditional Chinese medicine, is sweet and warm in nature, nourishing qi and promoting yang, stabilizing the surface and stopping sweating. It has anti-tumor, cardiovascular and cerebrovascular, immune function, lung function, kidney tissue, liver injury, intestinal function, blood pressure regulation, anti-aging, and osteoporosis prevention and treatment effects; The medicinal herb Atractylodes macrocephala has a sweet and slightly warm taste, which can invigorate the spleen and qi, dry dampness, and promote diuresis. Atractylodes macrocephala can help strengthen the function of nourishing qi; Windproof is a supplement for dispelling wind and relieving pain, as well as suppressing dampness and relieving pain. Huangqi combined with Atractylodes macrocephala prevents sweat from leaking out and makes it difficult for external pathogens to invade internally. This formula mainly uses Qi tonifying drugs, combined with small amounts of wind dispelling and surface relieving products to prevent wind damage and promote the absorption of nutrients. Commonly used in clinical practice to treat Qi deficiency constitution, effectively reduce the frequency of recurrent upper respiratory tract infections, pediatric asthma, chronic urticaria, eczema, and liver cancer.

At present, there are some literature reports on the quality analysis of Yupingfeng Powder, but the quality of the decoction pieces is affected by the place of origin, processing methods, etc., and there are also certain differences in clinical efficacy. Therefore, in this study, HPLC fingerprint spectra of 15 batches of Yupingfeng Powder were established using HPLC method, and the contents of 11 components in Yupingfeng Powder were simultaneously determined. At the same time, the quality was comprehensively evaluated by combining chemometric methods, providing a certain basis for establishing quality standards for Yupingfeng Powder decoction, elucidating the pharmacological mechanism of Yupingfeng Powder, and guiding rational clinical drug use.

When optimizing the extraction method of Yupingfeng Powder test solution, this experiment compared three extraction methods: ultrasound, reflux, and continuous reflux. The number of chromatographic peaks, separation degree, and extraction rate of 11 target components quantitatively determined in the Yupingfeng Powder chromatogram were analyzed as indicators. The RSD of the relative retention time of the chromatographic peaks was less than 3.0%, and there was a significant difference in the total extraction rate. Among them, ultrasound and extraction methods obtained higher substance content, but extraction time was longer. Finally, ultrasound method was comprehensively considered for extraction.

In the experiment, different mobile phase systems such as methanol water and acetonitrile water were investigated. The results showed that acetonitrile water had less interference and better separation effect, shorter analysis time, and a stable baseline during the gradient elution process. Therefore, acetonitrile water was selected as the mobile phase.

Comparing the chromatograms of 11 compounds detected at five wavelengths of 230, 254, 276, 300, and 320 nm, it was found that Huangqi had relatively more characteristic peaks detected at the 230 nm wavelength, with high peak separation and maximum absorption; The number of characteristic peaks detected by Fangfeng at detection wavelengths of 230 and 254 nm is basically the same; Atractylodes macrocephala has the maximum absorption at a detection wavelength of 230nm, while Atractylodes macrocephala I and Atractylodes macrocephala III have weaker absorption at other wavelengths. Therefore, 230nm was used as the detection wavelength.

This experiment established HPLC fingerprint spectra for 15 batches of Yupingfeng Powder. The main quantitative indicators for content determination were glucosinolates, isoflavones, mangiferin, and mangiferin in Astragalus membranaceus, as well as sesquiterpenoid glycosides, sesquiterpenoid glycosides, 5-O-methylvisamminol glycosides, and paeonol glycosides in Windproof, and Atractylodes macrocephala III, Atractylodes macrocephala I, and Atractylodes macrocephala II in Atractylodes macrocephala. Stoichiometric methods such as HCA, PCA, and OPLS-DA were used to investigate the integrity and differences of different batches of Yupingfeng Powder. The results showed that the similarity of HPLC fingerprint spectra of the 15 batches of samples ranged from 0.835 to 0.965, The results of HCA and PCA analysis are basically consistent, and the samples are divided into two categories. Combined with the information of the origin of the decoction pieces, it is found that the decoction pieces in the first category mostly come from Inner Mongolia and other regions, while the decoction pieces in the second category mostly come from Zhejiang and other regions. There are significant differences in soil, climate, and cultivation methods between the north and south, and the decoction pieces in each pharmacy come from different decoction piece processing factories in different origins. Authentic Chinese herbal medicines and excellent origins are the key factors affecting the quality of decoction pieces. The effective ingredients contained in Chinese herbal medicines vary due to different growth environments. Therefore, there is a certain correlation between the quality of decoction pieces and the production of traditional Chinese medicine decoction pieces, including the origin of the medicinal materials, the season of harvesting and processing, and the processing and manufacturing. The quality evaluation method established by our research institute is simple and has good repeatability, which can provide reference for drug manufacturers in the process of raw materials and preparation, and is conducive to the clinical rational use of Yupingfeng Powder.