This study aimed to develop an ultra-performance liquid chromatography(UPLC) method for the quantitative analysis of xanthones and oligosaccharide esters in Polygalae Radix(PR), and to explore a retention-time-ratio-based strategy for inter-column method transfer. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) by gradient elution using 0.1% formic acid in water and acetonitrile with detection at 238 nm. The UPLC fingerprints of 31 batches of PR samples were established, with 21 common peaks marked and identified by UPLC-Q-TOF-MS~E. Through systematic methodological validation, a quantitative method was developed for the simultaneous determination of 7 bioactive components, namely sibiricose A_1, sibiricaxanthone B, polygalaxanthone Ⅷ, polygalaxanthone Ⅺ, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose, and tenuifoliside A. A quantitative analysis of multiple components by single marker(QAMS) method was established using polygalaxanthone Ⅲ as the internal reference substance, and its accuracy was compared with that of the external standard method(ESM). The feasibility of method transfer across different columns and instruments was also investigated. All 7 analytes showed good linearity(r>0.999) within their respective concentration ranges with mean recoveries of 94.31%-99.02%(RSD<2.0%, n=6). The relative error(RE) between QAMS and ESM was less than 4.0%, and the paired t-test indicated no significant difference between the two methods. Preliminary studies of method transfer on 9 C_(18) columns from 3 manufacturers on Waters UPLC systems of two models demonstrated that gradient time adjustment based on retention time ratio could maintain the elution behavior and separation characteristics of the target peaks, though the generalizability of this strategy requires further validation. The established UPLC-QAMS method is accurate and reliable for the quality control of PR. The inter-column method transfer strategy provides preliminary reference for the broader application of analytical methods.

Poria cocos is a commonly used bulk traditional Chinese medicinal material, with multiple parts used medicinally, such as Poriae Cutis, Rubra Poria, White Poria, and Poria cum Radix Pini. Polysaccharides are their main active components. Clarifying the composition and content differences of polysaccharides in different medicinal parts of P. cocos can provide a reference for processing and clinical application of P. cocos medicinal materials and decoction pieces. In this paper, high-performance gel permeation chromatography(HPGPC) and nuclear magnetic resonance(NMR) spectroscopy were employed to qualitatively compare the relative molecular mass distribution and chemical composition of alkali-soluble polysaccharides from different parts of crude P. cocos. The results showed that the composition of alkali-soluble polysaccharides in Rubra Poria and Poriae Cutis was consistent with that in White Poria, all of which were β-(1→3)-D-glucans. The relative molecular mass distribution was also relatively close, ranging approximately from 9.13×10~4 to 1.04×10~5. On the basis of the content determination method of β-(1→3)-D-glucan in P. cocos according to the 2025 edition of Chinese Pharmacopoeia, the preparation method of the test sample solution was optimized. Firstly, a desalination step was added, which can enable the test sample chromatogram to present only the chromatographic peak of glucose, thereby reducing the difficulty of separation. Secondly, this experiment discovered for the first time that under acidic conditions, the hydrolysis of β-(1→3)-D-glucan to glucose was accompanied by the generation of a small amount of 5-hydroxymethylfurfural(5-HMF) as a by-product. A more intense acid hydrolysis reaction condition indicated a greater conversion yield of 5-HMF. Therefore, according to the results of single-factor experiments and response surface methodology experiments, the acid hydrolysis time was shortened from 6 h to 1.5 h, which improved the analysis efficiency. After verification, the optimized method was simple and stable, and had good repeatability and accuracy. Then, the optimized quantitative method was used to evaluate the content of β-(1→3)-D-glucan in different parts of crude P. cocos and decoction pieces. The results demonstrated that the content of β-(1→3)-D-glucan in different medicinal parts of P. cocos was in the order of White Poria, Poria cum Radix Pini, Rubra Poria, and Poriae Cutis. Among the commercially available Poria decoction pieces, the content of β-(1→3)-D-glucan in the white and hard decoction pieces was higher than that in the dark and loose-textured decoction pieces.

Alisma Rhizome (AR) is widely used in China to treat hyperlipidaemia and diabetes, with polysaccharides being the main components of AR aqueous extract. However, their detailed structural characterisation remains unclear. In this study, six purified polysaccharides (AP-1a, AP-1b, AP-1c, AP-2a, AP-3a, and AP-4a) were isolated from AR. Structural characterisation revealed that AP-1a (150.8 kDa) featured a 1,4-linked α-D-Glcp backbone with α-D-Glcp terminal groups at C-6 for every nine 1,4-α-D-Glcp units. AP-1b (18.13 kDa) and AP-1c (8.49 kDa) had a 1,4-linked α-D-Glcp and 1,4-linked β-D-Galp backbone in an 8:1 ratio, with minor t-α-D-Glcp at C-6 for every eight 1,4-α-D-Glcp units. AP-2a (27.28 kDa) possessed a → 3,6)-β-D-Galp-(1 → 3)-β-D-Galp-(1 → repeating backbone with side chains attached to C-6 of →3,6)-β-D-Galp-(1→. The backbone of AP-3a (34.40 kDa) was composed of →3)-β-D-Galp-(1→, →3,6)-β-D-Galp-(1 → and →2,4)-α-D-Rhap-(1 → in a 4:4:1 ratio, and the side chains substituted at C-6 of →3,6)-β-D-Galp-(1 → and C-4 of →2,4)-α-D-Rhap-(1→, respectively. AP-4a (24.49 kDa) had 1,4-linked α-D-GalAp as the main chain, and was branched at C-3 by t-α-D-GalAp units. All of the above polysaccharides activated nitric oxide and tumour necrosis factor-α secretion in macrophages, indicating significant immunomodulatory activities. AP-3a and AP-4a exhibited more significant antioxidant activities in free radical scavenging assays and H2O2-induced HepG2 cells. Moreover, AP-3a reduced cholesterol and triglyceride levels in oleic acid-induced HepG2 cells. Results suggest that AR polysaccharides have good immunomodulatory, antioxidant and hypolipidemic activities in vitro.

Despite that the leaves of Ligustrum lucidum Ait. (LLA) have been used in traditional Chinese medicine for more than 2000 years, their medicinal potential remains underexplored compared with the fruits of this plant species. Therefore, the aims of this study were to investigate the antioxidative activities of different solvent fractions of LLA leaf extract and to identify the active components and their spatial distribution within the leaf tissue. Among all solvent fractions tested, the ethyl acetate and n-butanol fractions showed the strongest antioxidative activities in the 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, and fluorescence recovery after photobleaching assays. In total, 19 and 21 compounds were identified in the ethyl acetate and n-butanol fractions, respectively, with flavonoids, iridoids, and triterpenes being predominant. Analysis using desorption electrospray ionization-post-photoionization-mass spectrometry imaging identified 20 compounds that were mainly distributed in the leaf epidermis and mesophyll. These findings provide a comprehensive reference resource that will aid our understanding of the medicinal value of LLA leaves and lay the scientific foundation for their further development and use as a source of natural antioxidants.

To compare the safety profiles of Gardeniae Fructus (Zhizi) and its counterfeit, Shuizhizi, Sprague-Dawley rats were orally administered aqueous extracts of Zhizi and Shuizhizi at 1.8 and 5.4 g/kg/day for 28 and 90 days, followed by a 28-day recovery period. Hematological indices, biochemical markers of liver and kidney function, organ coefficients, and histopathology were assessed at all three time points. Both substances induced dose-dependent abnormal hematological findings, disrupted liver/kidney function indices, altered organ coefficients, and caused histopathological damage. Shuizhizi induced significantly more severe toxicities than Zhizi. After the recovery period, the safety gap widened, with the Shuizhizi groups exhibiting delayed recovery: significantly reduced body weight in the high-dose Shuizhizi group (H-SZZ), significant alterations in most hematology and liver/kidney indices in both the low-dose Shuizhizi group (L-SZZ) and H-SZZ groups, and persistent marked liver pathology in the L-SZZ group. These findings indicate that Zhizi and Shuizhizi exhibit an identical spectrum of toxicity. However, the latter displayed significantly greater toxicity, with residual toxicity being particularly pronounced during the recovery phase. These results confirm that Shuizhizi is less safe for clinical application compared to Zhizi and should not be used as a substitute in clinical practice.

In chrysanthemums, flower color is an important market trait that depends on carotenoid composition and contents. Although polyphenols have been widely reported, carotenoids have rarely been systematically investigated in chrysanthemums. Here, 68 carotenoids in 12 representative chrysanthemum cultivars were quantitatively analyzed using comparative metabolomics to explore the mechanism of carotenoid accumulation. Yellow chrysanthemums contained more antheraxanthin, violaxanthin, and lutein than other chrysanthemums. Purple chrysanthemums primarily contained astaxanthin, lycopene, and phytofluene. These compounds might be used as carotenoid markers to differentiate yellow from purple chrysanthemums. Correlation analysis of differential carotenoids and color revealed that b* value was positively correlated with most carotenoids, particularly carotene and lutein derivatives. Furthermore, the high expression of PSY-related genes in yellow chrysanthemums provided enough phytoene as precursors for generating downstream carotenoids, while CHYB, ZEP, and LUT5-related genes promoted the synthesis of the corresponding carotenoids, contributing to high carotenoid contents in yellow chrysanthemums. This study is the first to establish a method to analyze multiple carotenoids in chrysanthemums, revealing the mechanism of carotenoid accumulation, and laying the foundation for molecular breeding of various colored chrysanthemums.

Chrysanthemum (Chrysanthemum morifolium Ramat) is a valuable crop mainly used for medicine, beverages and ornament. Enzymatic browning causes serious postharvest flower deterioration of chrysanthemum, but the mechanism remains unclear. To clarify the mechanism involved, the cellular ultrastructure, metabolites, browning products, and differentially expressed genes of chrysanthemum were studied systematically. During the browning process, cellular compartmentalization was severely disrupted and the contents of chlorogenic acid derivatives and flavonoid glycosides decreased significantly. A browning product with a dihydrobenzofuran skeleton was purified and identified for the first time from a simulated browning reaction of chlorogenic acid catalyzed by peroxidase. Moreover, six differentially expressed genes were for the first time identified to be involved in the regulation of oxidase by transcriptome and qPCR, and one of which (c76073_g1) was validated using in vitro enzyme activity determination. This research elucidates the inherent mechanism of enzymatic browning, provides a scientific explanation for the steam-treated processing of chrysanthemums and lays the foundation for the molecular breeding of anti-browning chrysanthemum varieties.

Ganoderma spores are the dry and mature spores of G. lucidum or G. sinense of Polyporaceae, with strong immunity-improving, anti-tumor, and anti-oxidant activities. Triterpenoids are one of the main active components in Ganoderma spores, while the method for determining the content of triterpenoids remains to be developed. In this paper, UPLC-Q-TOF-MS/MS was employed to qualitatively analyze the triterpenoids in G. lucidum spores, and an HPLC method was established for simultaneous determination of five triterpenoids in the spores. The comparison on the retention time, ultraviolet spectrum, and fragmentation with the reference substance in the negative ion mode revealed nine triterpenoids in G. lucidum spores. These nine triterpenoids were ganoderic acid I, ganoderenic acid C, ganoderic acid C2, ganoderic acid C6, ganoderic acid G, ganoderenic acid B, ganoderic acid B, ganoderic acid A, and ganoderic acid H. Three triterpenoids from the spores were inferred by comparison with the available articles, including lucidenic acid E,(Z)-6-\[(3S,5R,7S,10S,12S,13R,14R,17R)-12-acetoxy-3,7-dihydroxy-4,4,10,13,14-pentamethyl-11,15-dioxo-2,3,4,5,6,7,10,11,12,13,14,15,16,17-tetrahydro-1H-cyclopenteno \[α\] phenanthrene-17-yl\]-2-methyl-4-oxoheptyl-5-enoic acid, and ganoderin A. With Ultimate HPLC XS-C_(18) column(4.6 mm×250 mm, 5 μm) as the stationary phase and acetonitrile-0.072% phosphoric acid solution as the mobile phase, gradient elution was carried out at a flow rate of 1.2 mL·min~(-1), detection wavelength of 257 nm, and column temperature of 40 ℃. These conditions enabled the simultaneously quantitative analysis of the five triterpenoids, and the results of methodological investigation showed that the method met the requirements of content determination. Ganoderenic acid C, ganoderic acid C2, ganoderic acid C6, ganoderenic acid B, and ganoderic acid H showed a good linear relationship(R~2>0.999) within the concentration ranges of 0.93-16.08, 3.44-23.88, 4.28-59.40, 1.40-16.16, and 3.53-61.20 μg·mL~(-1), respectively, with the average recovery rate of 79.53%-93.60% and RSD of 4.7%-7.6%. The content of the five triterpenoids in 15 batches of raw spores and 4 batches of sporoderm-broken spores from different habitats was within the ranges of 0.44-16.25 μg·g~(-1) and 0.52-3.57 μg·g~(-1), respectively, which showed a large difference. The method established in this paper was simple and reliable and could be used for the accurate determination of triterpenoids in G. lucidum spores, which provides a methodological basis for the quality evaluation of G. lucidum spores.

Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to rapidly identify the chemical components in Dracocephalum moldavica, and UPLC was employed to determine the content of its main components. MS analysis was performed using an electrospray ionization(ESI) source and data were collected in the negative ion mode. By comparing the retention time and mass spectra of reference compounds, and using a self-built compound database and the PubChem database, 68 compounds were identified from D. moldavica, including 36 flavonoids, 22 phenylpropanoids, 4 phenols, and 6 other compounds. On this basis, a UPLC quantitative method was established to simultaneously determine 8 main components, i.e., luteolin-7-O-glucuronide, apigenin-7-O-glucuronide, rosmarinic acid, diosmetin-7-O-glucuronide, tilianin, acacetin-7-O-glucuronide, acacetin-7-O-(6″-O-malonyl)-glucoside, and acacetin. A Waters ACQUITY BEH C_(18) column(2.1 mm × 100 mm, 1.7 μm) was used, with acetonitrile and a water solution containing 0.1% formic acid and 0.1% phosphoric acid as the mobile phase for gradient elution. The detection wavelength was set at 330 nm, with a flow rate of 0.4 mL·min~(-1), and the column temperature was maintained at 35 ℃. The 8 components demonstrated good linearity(r≥0.999 9) over a wide mass concentration range(50 or 100 times). The average recovery rate ranged from 97.5% to 105.1%, and the relative standard deviations(RSDs) were 0.90% to 3.4%(n= 6), indicating that the method was simple, accurate, and reliable. In 17 batches of D. moldavica samples, the content of these 8 components ranged from 0.405 to 2.10, 0.063 to 0.342, 0.446 to 2.43, 0.415 to 1.47, 1.57 to 4.34, 0.173 to 0.386, 1.00 to 5.40, and 0.069 to 0.207 mg·g~(-1), respectively. These results indicate significant differences in the internal quality of the samples, highlighting the need for strict quality control to ensure their pharmacodynamic efficacy. This study provides a scientific basis for the rapid discovery of pharmacodynamic substances, comprehensive quality control, and the formulation or revision of quality standards for D. moldavica.

目的：建立杜仲叶药材中5种成分（桃叶珊瑚苷、京尼平苷酸、绿原酸、车叶草苷、芦丁）含量测定的一测多评法（QAMS），并验证其在杜仲叶药材含量测定中的可行性和适用性，为杜仲叶药材质量标准的制定提供科学依据。方法：采用高效液相色谱法（HPLC），Welch Boltimate? C 18 色谱柱（4.6 mm×100 mm，2.7 μm），以甲醇（A）-0.2%磷酸水溶液（B）为流动相进行梯度洗脱（0～8 min，3%A；8～10 min，3%～11%A；10～26 min，11%A；26～27 min，11%～25%A；27～60 min，25%～32%A），流速0.6 mL·min -1 ，柱温30 ℃，检测波长210 nm和254 nm。以绿原酸为内参物，建立与其他4个指标成分之间的相对校正因子(f s/i )，并分别采用QAMS和外标法（ESM）测定14批杜仲叶药材中5种成分的含量。结果：桃叶珊瑚苷、京尼平苷酸、车叶草苷和芦丁的f s/i 分别为3.13，1.45，2.64，0.56，在不同实验条件下重复性良好，相对标准偏差（RSD）在0.5%～4.9%。测得14批杜仲叶中桃叶珊瑚苷、京尼平苷酸、绿原酸、车叶草苷、芦丁的质量分数分别为1.340～28.975，0.252～36.086，10.016～27.443，1.396～8.646，0.533～1.766 mg·g -1 ，QAMS与ESM比较，结果无明显差异（RSD＜5.0%）。结论：以绿原酸为内参物建立的QAMS可用于杜仲叶药材中5种成分的含量测定，该方法简便易行、结果准确。经综合评估，建议绿原酸（不低于1.5%），桃叶珊瑚苷（不低于1.0%）及京尼平苷酸（不低于1.0%）作为杜仲叶的质量控制标准，可较好地避免因2005—2020年版《中国药典》指标成分（绿原酸）专属性差和含量限度过低而带来的潜在质量风险。

Prunellae Spica is the dried spica of Prunella vulgaris belonging to Labiatae and it is widely used in pharmaceutical and general health fields. As a traditional Chinese medicine cultivated on a large scale, it produces a large amount of non-medicinal parts, which are discarded because they are not effectively used. To analyze the chemical constituents in the different samples from spica, seed, stem, and leaf of P. vulgaris, and explore the application value and development prospect of these parts, this study used ultrahigh performance liquid chromatography-tandem quadrupoles time of flight mass spectrometry(UPLC-Q-TOF-MS/MS) to detect chemical constituents in different parts of P. vulgaris. As a result, 117 compounds were detected. Among them, 87 compounds were identified, including 32 phenolic acids, 8 flavonoids, and 45 triterpenoid saponins. Some new triterpenoid saponins containing the sugar chain with 4-6 sugar units were found. Further, multivariate statistical analysis was conducted on BPI chromatographic peaks of multiple batches of different parts, and the results showed that spica had the most abundant chemical constituents, including salviaflaside and linolenic acid highly contained in the seed and phenolic acids, flavonoids, and triterpenoid saponins in the stem and leaf. In general, the constituents in the spica were composed of those in the seed, stem, and leaf. UPLC was used to determine the content of 6 phenolic acids(danshensu, protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside, and rosmarinic acid) in different parts. The content of other phenolic acids in the seed was generally lower than that in the spica except that of salviaflaside. The content of salviaflaside in the spica was higher than that in the stem and leaf, but the content of other phenolic acids in the spica was not significantly different from that in the stem. The content of protocatechuic aldehyde and caffeic acid in the spica was lower than that in the leaf. DPPH free radical scavenging method was used to detect the antioxidant activity of four parts, and there was no significant difference in the antioxidant activity between the spica and the stem and leaf, but that was significantly higher than the seed. Moreover, the antioxidant activity of these parts was correlated with the content of total phenolic acids. Based on the above findings, the stem and leaf of P. vulgaris have potential application value. Considering the traditional medication rule, it is feasible to use the whole plant as a medicine. Alternatively, salviaflaside, occurring in the seed, can be used as a marker compound for the quality evaluation of Prunellae Spica, if only using spica as the medicinal part of P. vulgaris, as described in the Chinese Pharmacopoeia(2020 edition).

Galli Gigerii Endothelium Corneum(GGEC), the dried gizzard membrane of Gallus gallus domesticus is a Chinese medicinal material commonly used for digestion. However, due to the particularity of texture and composition, its active ingre-dients have not been clarified so far, and there is also a lack of quality evaluation indicators. In this study, UPLC-Q-TOF-MS was used to analyze the chemical components from the water extract of GGEC, and ten nucleosides were identified for the first time. HPLC fingerprints of the water extracts of GGEC were established and the content of seven nucleosides was determined. The fingerprint similarities of 40 batches of GGEC samples ranged from 0.765 to 0.959, indicating that there were great differences among the GGEC products processed with different methods. In addition, SPSS 22.0 and SIMCA 14.1 were used for hierarchical cluster analysis(HCA) and principal component analysis(PCA) on the 19 common peaks of the HPLC fingerprints of GGEC, and the 40 batches of samples were divided into three categories: raw GGEC, fried GGEC and vinegar-processed GGEC. Eight differential components in GGEC were marked by orthogonal partial least squares discrimination analysis(OPLS-DA), two of which were adenine and thymine. The results of content determination showed that the total content of the seven nucleosides in raw GGEC, fried GGEC and vinegar-processed GGEC were 182.5-416.8, 205.3-368.7, and 194.2-283.0 μg·g~(-1), respectively. There were significant differences in the content of hypoxanthine, thymine and thymidine among the GGEC products processed with different methods(P<0.05), which were graded in the order of fried GGEC>vinegar-processed GGEC>raw GGEC. This suggested that the content of hypoxanthine, thymine and thymidine tended to increase during the frying process, and the variation range might be related to the degree of heat exposure. The established methods in this study were simple and reproducible, and could be used for qualitative and quantitative analysis of GGEC and its processed pro-ducts. This study also provided reference for the establishment of quality standards of GGEC with chemical components as control index.

Rosae Radix et Rhizoma is a herbal medicine in a variety of famous Chinese patent medicines, while the quality standard for this medicine remains to be developed due to the insufficient research on the quality of Rosae Radix et Rhizoma from different sources. Therefore, this study comprehensively analyzed the components in Rosae Radix et Rhizoma of different sources from the aspects of extract, component category content, identification based on thin-lay chromatography, active component content determination, and fingerprint, so as to improve the quality control. The results showed that the content of chemical components varied in the samples of different sources, while there was little difference in the chemical composition among the samples. The content of components in the roots of Rosa laevigata was higher than that in the other two species, and the content of components in the roots was higher than that in the stems. The fingerprints of triterpenoids and non-triterpenoids were established, and the content of five main triterpenoids including multiflorin, rosamultin, myrianthic acid, rosolic acid, and tormentic acid in Rosae Radix et Rhizoma was determined. The results were consistent with those of major component categories. In conclusion, the quality of Rosae Radix et Rhizoma is associated with the plant species, producing area, and medicinal parts. The method established in this study lays a foundation for improving the quality standard of Rosae Radix et Rhizoma and provides data support for the rational use of the stem.

In order to establish the standardized processing technology of the hot water washing of Euodiae Fructus, this study, based on the traditional processing method of hot water washing of Euodiae Fructus recorded in ancient works and modern processing specifications of traditional Chinese medicine decoction pieces, took the yield of decoction pieces and the content of main components as the indicators and optimized the processing conditions by orthogonal test based on the results of single factor investigation. At the same time, electronic tongue technology was used to analyze the change law of the taste index of Euodiae Fructus during the hot water washing. The results of the single factor investigation showed that the content of the main components in Euodiae Fructus showed some regular changes during the processing. Specifically, the content of chlorogenic acid, hyperin, isorhamnetin-3-O-rutinoside, isorhamnetin-3-O-galactoside, and dehydroevodiamine decreased significantly, with average decreases of-23.75%,-27.80%,-14.04%,-14.03%, and-13.11%, respectively. The content of limonin increased significantly with an average increase of 19.83%. The content of evodiamine, rutaecarpine, evocarpine, and dihydroevocarpine showed fluctuating changes and generally increased, with average variation amplitudes of 0.54%,-3.78%, 2.69%, and 5.13%, respectively. The orthogonal test results showed that the optimum processing parameters for the hot water washing of Euodiae Fructus were as follows: washing time of 2 min, the solid-to-liquid ratio of 1∶10 g·mL~(-1), washing temperature of 80 ℃, washing once, and drying at 50 ℃. After the hot water washing processing, the average yield of Euodiae Fructus pieces was 94.80%. The content of limonin, evodiamine, and rutaecarpine was higher than those of raw pro-ducts, and the average transfer rates were 102.56%, 103.15%, and 105.16%, respectively. The content of dehydroevodiamine was lower than that of the raw products, and the average transfer rate was 83.04%. The results of taste analysis showed that the hot water washing could significantly reduce the salty, astringent, and bitter tastes of Euodiae Fructus. This study revealed the influence of the hot water washing on the content of main components and taste of Euodiae Fructus, and the processing technology of the hot water was-hing of Euodiae Fructus established in this study was stable, feasible, and suitable for industrial production, which laid a foundation for clarifying its processing principle and improving the quality standard and clinical application value of decoction pieces.

With the improvement of living standards and changes in working style, the prevalence of abnormal glucose and lipid metabolism in humans is increasing in modern society. Clinically, the related indicators are often improved by changing the lifestyle and/or taking hypoglycemic and lipid-lowering drugs, but there are no therapeutic drugs for disorders of glucose and lipid metabolism at present. Hepatitis C virus core protein binding protein 6(HCBP6) is a newly discovered target that can regulate triglyceride and cholesterol content according to level oscillations in the body, thereby regulating abnormal glucose and lipid metabolism. Relevant studies have shown that ginsenoside Rh_2 can significantly up-regulate the expression of HCBP6, but there are few studies on the effect of Chinese herbal medicines on HCBP6. Moreover, the three-dimensional structural information of HCBP6 has not been determined and the discovery of potential active components acting on HCBP6 is not rapidly advanced. Therefore, the total saponins of eight Chinese herbal medicines commonly used to regulate abnormal glucose and lipid metabolism were selected as the research objects to observe their effect on the expression of HCBP6. Then, the three-dimensional structure of HCBP6 was predicted, followed by molecular docking with saponins in eight Chinese herbal medicines to quickly find potential active components. The results showed that all total saponins tended to up-regulate HCBP6 mRNA and protein expression, where gypenosides showed the optimum effect on up-regulating HCBP6 mRNA and ginsenosides showed the optimum effect on up-regulating HCBP6 protein expression. Reliable protein structures were obtained after the prediction of protein structures using the Robetta website and the evaluation of the predicted structures by SAVES. The saponins from the website and literature were also collected and docked with the predicted protein, and the saponin components were found to have good binding activity to the HCBP6 protein. The results of the study are expected to provide ideas and methods for the discovery of new drugs from Chinese herbal medicines to regulate glucose and lipid metabolism.

To improve the quality control methods of Poria and develop and utilize its resources fully, alkaline extraction was used in this study to determine the yield and content of alkali-soluble polysaccharides of Poria. The alkali-soluble extracts of Poria were obtained according to the optimum extraction conditions on the basis of single-factor test, and 30 batches of samples were determined. The structure and chemical composition of the alkali-soluble extracts was characterized by high-performance gel permeation chromatography(HPGPC), Fourier transform infrared spectrometry(FT-IR), nuclear magnetic resonance(NMR) spectroscopy and high-performance liquid chromatography(HPLC) with 1-phenyl-3-methyl-5-pyrazolone(PMP-HPLC). The results showed that the content of the alkali-soluble extracts was in the range of 46.98%-73.86%. The main component was β-(1→3)-glucan, and its molecular mass was about 1.093×10~5. Further, the content of alkali-soluble polysaccharides of Poria was measured by UV-Vis spectrophotometry and HPLC coupled with the evaporative light scattering detector(HPLC-ELSD), and 30 batches of samples were measured. The results indicated that the content of alkali-soluble polysaccharides determined by UV-Vis spectrophotometry was in the range of 73.70%-92.57%, and the content of samples from Hubei province was slightly higher than that from Yunnan province, Anhui province and Hunan province. The content of alkali-soluble polysaccharides determined by HPLC-ELSD was in the range of 51.42%-76.69%, and the samples from Hunan province had slightly higher content than that from the other three provinces. The content determined by UV-Vis spectrophotometry was higher than that by HPLC-ELSD. However, the content determined by HPLC-ELSD was close to that of alkali-soluble extract, which could accurately characterize the content of alkali-soluble polysaccharides in Poria, and the method was simple and repeatable. Therefore, it is recommended that the quantitative analysis method for alkali-soluble extract and alkali-soluble polysaccharides by HPLC-ELSD be used in the quality standards of Poria in Chinese Pharmacopeia.

To explore the stability characteristics of β-nicotinamide mononucleotide(NMN)and provide data support for NMN production,preparation,and related product development,this study established a simple HPLC content determination method for NMN in simple substrate and investigated the degradation behavior,degradation products,and degradation kinetics of NMN under various chemical,physical,and biological conditions.The HPLC method employed a Welch Xtimate AQ-C18 column(4.6 mm×250 mm,5 μm),a detection wavelength of 266 nm,a column temperature of 30℃,a flow rate of 1.0 mL·min-1,an injection volume of 5 μL,and a mobile phase consisting of methanol(A)and a 10 mmol·L-1 ammonium formate aqueous solution(B)with a gradient elution(0-6.7 min,0-4%A;6.7-13 min,4%-18%A;13-14.2 min,18%A;14.2-15 min,18%-0 A;15-22 min,0 A).This method provided good separation between NMN and potential impurities and degradation products,and had a wide linear range,short analysis time,good durability,high accuracy,an average sample recovery rate of 98.71%,and an RSD of 1.2%.The instrument precision had an RSD of 0.26%,and the linearity within the examined range was excellent(R2≥0.999 9).This method can be applied for NMN content determination in simple substrate.The degradation process of NMN in aqueous solution followed apparent first-order kinetics,with the degradation rate primarily influenced by high temperature and pH.NMN was more stable in low-temperature,neutral,or weakly acidic/alkaline environments.Strong acids or strong alkalis could accelerate its degradation,and its degradation rate was less affected by pepsin and trypsin.In an aqueous solution at room temperature,it followed the kinetic equation lg Ct=0.005 7t + 4.817 2,with t0.9 and t1/2 values of 95.58,860.26 h,respectively.The results suggest that pH and temperature are the main factors affecting the stability of NMN in aqueous solution,and low temperature,moisture protection,and a weakly acidic environment are more conducive to the storage and application of NMN and its products.

The fruits of Gardenia jasminoides Ellis are an important herb medicine in Traditional Chinese Medicine (TCM) and have been used for thousands of years for clearing away heat and toxic materials. It mainly contains iridoids, pigments, organic acids, and flavonoids. Although belonging to one species, it has two kinds of cultivars and one variety widely distributed and sold. This study aims to develop an integrated and efficient analytical strategy for comprehensive profiling of phytochemicals and clarify the differences in all three populations. Based on reversed-phase ultra-high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC/ESI-QTOFMS), an optimized analytical approach for comprehensive profiling of phytochemicals in the fruits of G. jasminoides was established in negative ionization mode. The holistic metabolites profiling was carried out on UHPLC/ESI-QTOFMS and data analysis program Progenesis QI, and a total of 80 metabolites were obtained and interpreted by chromatographic and tandem mass spectral data. The interpretation of metabolites comprises iridoids, pigments, organic acids, and flavonoids. Principal component analysis and partial least square-discriminant analysis were performed, and 19 main different components could be obtained to distinguish the three populations. Combined with non-targeted and targeted data analysis, the integrated strategy developed in this study was feasibly applied to discern differences in the profiles of the phytochemicals accumulating in the fruits of three populations of G. jasminoides.

A comprehensive quality control method was established to provide references for quality control and evaluation of substance benchmarks of Danggui Sini Decoction(DSD). The HPLC separation was performed on a Kromasil 100 C-8 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.05% phosphoric acid in water(B) as mobile phase in a gradient elution mode at the flow rate of 1 mL·min~(-1). The column temperature was 25 ℃ and the detection wavelength was set at 275 nm. Under these conditions, the content of seven components, including paeoniflorin, liquiritin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate, ligustilide, and asarinin was simultaneously determined. Under the same chromatographic conditions, the HPLC fingerprint method for analysis of 15 batches of DSD was established. The content determination of aristolochic acid I, using the same test solution as the content determination item, was performed on an ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) with methanol(A)-water(including 0.1% formic acid and 5 mmol·L~(-1) ammonium formate)(B) as the mobile phase in a gradient elution mode at the flow rate of 0.4 mL·min~(-1) and the column temperature of 40 ℃ by LC-MS/MS. The MS conditions included electrospray ionization(ESI) as an ion source, positive ion ionization, selective reaction monitoring(SRM), the parent ion of 359.3, and the daughter ion of 297.8. The results of the methodological investigation all met the requirements of content determination/fingerprint determination. As a result, the content ranges of paeoniflorin, liquiritin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate, ligustilide, and asarinin were 5.419 8-11.267 3, 1.023-3.669 8, 0.145 6-0.444 1, 0.099 1-0.321 9, 3.159 1-7.731 9, 0.146 4-0.471 7, and 0.237 3-0.401 0 mg·g~(-1), respectively. Twenty-two common peaks were selected and 10 of them were identified by the comparison with the reference substances. The fingerprint similarity of 15 batches of DSD was in the range of 0.91-0.996 and the content of aristolochic acid I in DSD was 300.03-638.13 ng·g~(-1). The method established in this study is reliable and easy to operate and has great practical value, which can be used for overall quality control of substance benchmarks for DSD.

Fifteen undescribed diterpenoid alkaloids and seven known analogs were coisolated from the whole plant Aconitum tanguticum (Maxim.) Stapf. Their structures were elucidated based on spectroscopic methods. Among them, tangirine A was a heteratisine-hetidine-type bis-diterpenoid alkaloid, tanguticinines A-F were six hetidine-hetisine-type bis-diterpenoid alkaloids, tanguticinine G was one hetidine-atisine-type bis-diterpenoid alkaloid, and N-oxide anthoroidine B, 5-deoxyanthoridine B and N-oxide 5-deoxyanthoroidine B were three unusual hetidine-rearranged hetisine-type bis-diterpenoid alkaloids. In the bioassay, thirteen compounds showed some inhibitory effects on the secretion of NO and TNF-alpha in LPS-treated RAW 264.7 cells, with IC50 values of 67.56 mu M-683.436 mu M.