Green and Efficient Extraction of Phenolic Components from Plants with Supramolecular Solvents: Experimental and Theoretical Studies.

The supramolecular solvent (SUPRAS) has garnered significant attention as an innovative, efficient, and environmentally friendly solvent for the effective extraction and separation of bioactive compounds from natural resources. However, research on the use of a SUPRAS for the extraction of phenolic compounds from plants, which are highly valued in food products due to their exceptional antioxidant properties, remains scarce. The present study developed a green, ultra-sound-assisted SUPRAS method for the simultaneous determination of three phenolic acids in Prunella vulgaris using high-performance liquid chromatography (HPLC). The experimental parameters were meticulously optimized. The efficiency and antioxidant properties of the phenolic compounds obtained using different extraction methods were also compared. Under optimal conditions, the extraction efficiency of the SUPRAS, prepared with octanoic acid reverse micelles dispersed in ethanol-water, significantly exceeded that of conventional organic solvents. Moreover, the SUPRAS method demonstrated greater antioxidant capacity. Confocal laser scanning microscopy (CLSM) images revealed the spherical droplet structure of the SUPRAS, characterized by a well-defined circular fluorescence position, which coincided with the position of the phenolic acids. The phenolic acids were encapsulated within the SUPRAS droplets, indicating their efficient extraction capacity. Furthermore, molecular dynamics simulations combined with CLSM supported the proposed method's mechanism and theoretically demonstrated the superior extraction performance of the SUPRAS. In contrast to conventional methods, the higher extraction efficiency of the SUPRAS can be attributed to the larger solvent contact surface area, the formation of more types of hydrogen bonds between the extractants and the supramolecular solvents, and stronger, more stable interaction forces. The results of the theoretical studies corroborate the experimental outcomes.

[Analysis of differential metabolites of spikes of Prunella vulgaris at different stages based on UPLC-MS/MS].

Prunella vulgaris, aptly named for its withering at the summer solstice, displays significant variation in quality arising from differing harvest time. However, research on the chemical composition changes of its spikes at various stages is limited, and the specific metabolites remain unclear. In order to elucidate the metabolites and metabolic pathways of the spikes of P. vulgaris, the current study deployed ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) and targeted metabolomics to characterize the compound variability in the spikes of P. vulgaris across different periods. Multivariate statistical techniques such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to identify the differences in metabolites, and relevant metabolic pathways were analyzed. A total of 602 metabolites were identified by metabolomics, of which organic acids and their derivatives were the most abundant, followed by flavonoids. Multiple differential metabolites, including p-hydroxybenzoic acids and gallic acids were identified based on variable importance in projection(VIP)>1 and P<0.05. The results of enrichment analysis suggested that isoflavonoids biosynthesis, aminobenzoate degradation, benzoate degradation, anthocyanins biosynthesis, metabolic pathways, microbial metabolism in different environments, secondary plant metabolite biosynthesis, tryptophan metabolism, and phenylpropanoid synthesis were the main metabolic pathways. These results intend to elucidate the dynamic changes of differential metabolites of P. vulgaris and provide a theoretical basis for further study of the harvesting mechanism of spikes of P. vulgaris.

Preparation and Characterization of Polysaccharides from Turpiniae Folium and Its Antioxidative, Anti-Inflammatory Activities and Antiproliferative Effect on VSMCs.

Turpiniae Folium, the dried leaves of Turpinia arguta Seem., is a kind of historic traditional Chinese medicine. Here, based on our previous study, we extracted the Turpiniae Folium polysaccharides (TFP) and isolated three polysaccharide fractions from TFP. Then, TFP and one of the major polysaccharide fractions (TFP-1a) were identified through HPLC, HPGPC, and ATR-FTIR. Furthermore, the evaluations of their antioxidative, anti-inflammatory activities and inhibitory effect on angiotensin II-induced vascular smooth muscle cells (VSCMs) proliferation in vitro were conducted. Both TFP and TFP-1a showed strong hydroxyl radical scavenging, DPPH radical scavenging, and Fe2+ chelating activities, and exerted strong anti-inflammatory activity. Moreover, TFP and TFP-1a also possessed a strong inhibitory effect on Ang II-induced VSCMs proliferation. On these premises, we inferred that TFP and TFP-1a could be potential and promising natural antioxidants, anti-inflammatory agents, and implicated to treat cardiovascular disease.

Discrimination and Prediction of Lonicerae japonicae Flos and Lonicerae Flos and Their Related Prescriptions by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy Combined with Multivariate Statistical Analysis.

LJF and LF are commonly used in Chinese patent drugs. In the Chinese Pharmacopoeia, LJF and LF once belonged to the same source. However, since 2005, the two species have been listed separately. Therefore, they are often misused, and medicinal materials are indiscriminately put in their related prescriptions in China. In this work, firstly, we established a model for discriminating LJF and LF using ATR-FTIR combined with multivariate statistical analysis. The spectra data were further preprocessed and combined with spectral filter transformations and normalization methods. These pretreated data were used to establish pattern recognition models with PLS-DA, RF, and SVM. Results demonstrated that the RF model was the optimal model, and the overall classification accuracy for LJF and LF samples reached 98.86%. Then, the established model was applied in the discrimination of their related prescriptions. Interestingly, the results show good accuracy and applicability. The RF model for discriminating the related prescriptions containing LJF or LF had an accuracy of 100%. Our results suggest that this method is a rapid and effective tool for the successful discrimination of LJF and LF and their related prescriptions.

[Differences of flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos based on headspace gas chromatography-ion mobility spectroscopy].

Lonicerae Japonicae Flos and Lonicerae Flos, as traditional Chinese medicinal and edible food, are widely used in medicine, food, health products, and other industries. However, there is no comprehensive study on the differences of flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. This study applied headspace gas chromatography-ion mobility spectrometry(HS-GC-IMS) to analyze the differences of flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. The differential biomarkers were confirmed by multivariate statistical analysis. The results showed that there were significant differences in the forty-seven flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. The differential markers were ethyl acetate, propyl alcohol, 1-octanol, 1-hexanol, hexanal, and(Z)-2-hexen-1-ol. Pathway enrichment analysis showed that the above markers were involved in the biosynthesis of major secondary metabolism, sulfate metabolism pathways, and formation of other flavor compounds. This study provides important references for the evaluation of flavor compounds of Lonicerae Japonicae Flos and Lonicerae Flos and the development of medicinal and edible products.

The therapeutic effects of Prunella vulgaris against fluoride-induced oxidative damage by using the metabolomics method.

Fluoride is considered as one of the most ubiquitous environmental pollutants. Numerous studies have linked reactive oxygen species (ROS)-dependent oxidative damage with fluoride intoxication, which could be prevented by antioxidants. However, the metabolomic changes induced by ROS disruptions in fluoride intoxication are yet unknown. The present study aimed to provide novel mechanistic insights into the fluoride-induced oxidative damage and to investigate the potential protective effects of ethanolic extract of Prunella vulgaris (natural antioxidant, PV) against fluoride-induced oxidative damage. The serum biochemical indicators related to fluoride-induced oxidative damage, such as lipid peroxidation parameter, inflammation and marker enzymes in the liver increased significantly in the fluoride-treated group, while antioxidant enzymes were decreased. However, PV treatment restored the level of these biochemical indicators, indicating satisfactory antioxidant, anti-inflammatory, and hepatoprotective potential of PV. The metabolomics analysis in the serum was performed by liquid chromatography-mass spectroscopy, whereas the fluoride treatment caused severe metabolic disorders in rats, which could be improved by PV. The differential metabolites screened by multivariate analysis after fluoride and PV treatment, were organic acids, fatty acids, and lipids. These differential metabolites represented disorders of glyoxylate and dicarboxylate metabolism and the citrate cycle (TCA) according to metabolic pathway analysis in fluoride treatment rats. Interestingly, the result of metabolic pathway analysis of post-treatment with PV was consistent with that of fluoride treatment, indicating that the energy metabolism plays a major role in the progress of fluoride-induced oxidative damage, as well as the therapeutic effect of PV. These findings provided a theoretical basis for understanding the mechanism underlying metabolic disorders of fluoride toxicity and the effect of PV.

[Metabolic mechanism of Prunella vulgaris in treatment of ethanol-induced oxidative stress in rats based on metabonomics].

Prunella vulgaris(PV) is an edible and traditional medicinal herb which has a wide range application in fighting inflammation and oxidative stress, and protecting liver. Now it has been used to treat various types of liver diseases and has significant clinical efficacy. This study aims to investigate the effects of PV on ethanol-induced oxidative stress injury in rats and its metabolic mechanism. The rats were divided into control group, model group, PV group, and VC group. The liver protection of PV was identified by measuring pharmacological indexes such as antioxidant and anti-inflammatory activity. The metabolic mechanism of long-term ethanol exposure and the metabolic regulation mechanism of PV treatment were studied by LS-MS metabonomics. The pharmacological investigation indicated that ethanol could significantly decrease the contents of SOD, GSH-Px, CAT and other antioxidant enzymes in liver and increase the content of MDA. At the same time, PV could significantly reduce the contents of inflammatory factors(TNF-α, IL-6 and IL-1ß) and liver function markers(ALT, AST, ALP) in serum. What's more, long-term ethanol exposure could significantly cause liver injury, while PV could protect liver. Metabolomics based on multiple statistical analyses showed that long-term ethanol exposure could cause significant metabolic disorder, and fatty acids, phospholipids, carnitines and sterols were the main biomarkers. Meanwhile, pathway analysis and enrichment analysis showed that the ß oxidation of branched fatty acids was the main influencing pathway. Also, PV could improve metabolic disorder of liver injury induced by ethanol, and amino acids, fatty acids, and phospholi-pids were the main biomarkers in PV treatment. Metabolic pathway analysis showed that PV mainly regulated metabolic disorder of ethanol-induced liver injury through phenylalanine, tyrosine and tryptophan biosynthetic pathways. This study could provide a new perspective on the hepatoprotective effect of natural medicines, such as PV.

Premature ovarian insufficiency may be associated with the mutations in mitochondrial tRNA genes.

Premature ovarian insufficiency (POI) is a common endocrine disorder featured by the triad constituting of amenorrhea for at least four months, to date, the molecular pathogenesis of POI is largely undetermined. Despite several investigations have reported an increase in reactive oxygen species (ROS) content in idiopathic POI, the role of mitochondrial DNA (mtDNA) mutations/variants in the progression of POI has not been widely investigated. The current study aimed to explore the association between mt-tRNA mutations/variants and POI; we first used the PCR-Sanger sequencing to detect the mutations/variants in mt-tRNA genes from 50 POI patients and 30 healthy subjects. In addition, we evaluated the mitochondrial functions by using trans-mitochondrial cybrid cells containing these potential pathogenic mt-tRNA mutations. Consequently, five mutations: tRNALeu(UUR) C3303T, tRNAMet A4435G, tRNAGln T4363C, tRNACys G5821A and tRNAThr A15951G were identified. Notably, these mutations occurred at the extremely conserved nucleotides of the corresponding mt-tRNAs and may result the failure in mt-tRNA metabolism and subsequently lead to the impairment in mitochondrial protein synthesis. Furthermore, biochemical and molecular analyses of the cybrid cells containing these mutations showed a significantly lower level of ATP production when compared with the controls, whereas the ROS levels were much higher in POI patients carrying these mt-tRNA mutations, strongly indicated that these mt-tRNA mutations may cause the mitochondrial dysfunction, and play active roles in the progression and pathogensis of POI. Together, this study shaded additional light on the molecular mechanism of POI that was manifestated by mt-tRNA mutations.

[Mechanisms of Fuke Qianjin Capsules in treatment of pelvic inflammatory disease based on GC-MS metabolomics].

Pelvic inflammatory disease( PID) rat model was induced by the mixture of Escherichia coli,Staphylococcus aureus,and Streptococcus hemolytic-ß． Gas chromatography-mass spectrometry( GC-MS) based metabolic profiling method was combined with multivariate statistical analysis,such as PCA,PLS-DA and OPLS-DA to analyze endogenous small molecule metabolites in serum of rats after treatment of Fuke Qianjin Capsules. The results showed that Fuke Qianjin Capsules could significantly improve the inflammatory pathological characteristics and tissue damages in model rats. Based on the principle of VIP>1 and P<0. 05,a total of 6 different metabolic biomarkers were identified,including L-valine,L-isoleucine,L-threonine,butanedioic acid,serine and D-glucose,respectively.The contents of these six different metabolites were significantly reversed after administration. Further analysis of the metabolite pathways through KEGG database showed that Fuke Qianjin Capsules achieved the effect possibly through glycine,serine and threonine metabolism,aminoacyl-tRNA biosynthesis and valine,leucine and isoleucine biosynthesis. Therefore,this study came to the conclusion that Fuke Qianjin Capsules can be used in the treatment of mixed bacteria induced pelvic inflammatory disease possibly by regulating amino acid and its derivative metabolism.

[Extraction of flavonoids in Prunella vulgaris based on deep eutectic solvent method：application of new green solvent].

Flavonoids have attracted much attention due to their good anti-inflammatory, anti-oxidation and anti-tumor effects. At present, the extraction of flavonoids is mainly based on organic solvent, while the researches on the use of green and safe solvents are quite limited. Therefore, in the present study, different types of deep eutectic solvents (DESs) were applied to investigate their effect on extraction of flavonoids and optimize the process, also investigate the recovery efficiency of DESs and evaluate the recovery method for total flavonoids. The extraction yield of the total flavonoids acted as the comprehensive evaluation indexes, and a central composite design (CCD) of response surface methodology (RSM) was employed to further optimize the alcohol-based DES extraction conditions. The results showed that the optimized extraction conditions were as follows: water-DES ratio of 27%, solid-liquid ratio of 15 mL·g⁻¹, extraction temperature of 83 °C and extraction time of 42 min in ChCl-glycerol at 1:4 ratio. Under these conditions, the mean experimental value of the extraction yield (75.05 mg·g⁻¹) corresponded well with the predicted value (77.86 mg·g⁻¹). Moreover, these experimental results showed more advantages such as in higher efficiency, economy and environmental protection as compared with previously reported conventional extraction methods. In addition，the recovery yield of the total flavonoids from the DESs extraction solution achieved 97.88% by using AB-8 macroporous resin, and 88.12% desorption ratio can be achieved by 100% ethanol with 5 times resin content. After the above treated DESs were collected, the extraction yield with the same method reached 95.23%, indicating that the method of macroporous resin can be used for efficient and simple recovery and reuse. This study suggests that DESs can be used as a kind of sustainable and efficient natural extraction solvents for extraction of flavonoids from Prunella vulgaris.

[Analysis on multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica based on multivariate statistical analysis].

Prunellae Spica is a perennial edible and medicinal plant, rich in antioxidant substances. Total flavonoids (TFC), Phenolics (TPC), triterpenoids (TSC), polysaccharides (PC) and their antioxidant capacities (by the FRAP, DPPH and ABTS⁺ methods) of ethyl acetate fraction, n-butanol fraction and other fractions of aqueous extract from Prunellae Spica were investigated in this study. Then the multivariate statistical method was adopted to analyze the relationship between the multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica. The results showed that ethyl acetate fraction had relatively high concentration of TFC (0.61±0.10) g·g⁻¹DW, TPC (0.52±0.09) g·g⁻¹DW, and TSC (0.21±0.03) g·g⁻¹DW, with high scavenging capacity of DPPH (3.1±0.38) mmol·L⁻¹·g⁻¹DW and FRAP (2.56±0.35) mmol·L⁻¹·g⁻¹DW. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) results indicated the information from chemical compositions and antioxidant capacity can represent the "differences" of different fractions. Canonical correlation analysis (CCorA) revealed a high positive correlation between the amounts of multiple chemical compositions and the antioxidant capacities (r=0.970 0), and the first canonical variate had been reached. Moreover, ABTS⁺ method showed a low response to the compositions of different fractions, so this method may not be suitable for evaluation of Prunellae Spica antioxidant capacities, while DPPH evaluation method was more suitable for TSC and TPC. The results of this study have important reference significance for the evaluation method on antioxidant activity of Prunellae Spica in the field of food or medicine as well as for the development of related extracts.

[Application of random forest algorithm in fingerprint of Chinese medicine: different brands of Xiasangju granules as example].

To establish a random forest algorithm for identifying and classifying different brands of Xiasangju granules, and provide effective reference for identifying multi-index complex fingerprint. HPLC method was used to collect the fingerprint of 83 batches of Xiasangju granules from different manufacturers. The classification of Xiasangju granules samples based on chromatographic fingerprints was identified by chemometric methods including principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA) and random forest analysis (RF). The superiority of the above three chemometric methods was compared. The results showed that the fingerprints of 83 batches of Xiasangju granules were established in this study. PCA could only explicate 56.52% variance contribution rate and could not completely classify the samples; PLS-DA analysis was superior to PCA, explicating 63.43% variance contribution rate and could obtain certain separation; RF could well classify the samples into 3 types, and the predication accuracy of the proposed method was 96.5%. Therefore, The results indicate that RF combined with HPLC fingerprint could effectively construct traditional Chinese medicine quality control and analysis system.

[Analyzing crude/processed root of Polygonum multiflorum from different habitats by UPLC fingerprint and mode identification methods].

To establish a content determination method for 2,3,5,4'-tetrahydroxy stilbene-2-O-ß-D-glucoside (TSG) of the crude/processed root of Polygonum multiflorum from different habitats in China and set up the fingerprint by using UPLC. Various samples were pretreated by macro-porous resin. Then UPLC analysis was performed on Waters ACQUITY UPLC@BEH C18 chromatographic column (2.1 mm×50 mm, 1.7 µm) at (25±5) ℃. A binary gradient elution system was composed of acetonitrile (phase A) and 0.5% acetic acid solution (phase B). Detection was performed at the wavelength of 254 nm, and the mobile flow rate was set at 0.3 mL•min⁻¹. Results showed that the yield of extraction of the 2,3,5,4'-tetrahydroxy stilbene-2-O-ß-D-glucoside from root of P. multiflorum was all over 25.0% after macro-porous resin separation; an exclusive UPLC fingerprint method of the crude/processed root of P. multiflorum from different habitats was successfully set up and 17 chromatographic peaks were calibrated. Cluster analysis can not entirely distinguish the crude one from the processed one, while principal component analysis absolutely can. 2,3,5,4'-tetrahydroxy stilbene-2-O-ß-D-glucoside is the composition that has largest differences in variable importance in projection (VIP) between crude and processed root of P. multiflorum. The separating method can gain high-purity 2,3,5,4'-tetrahydroxy stilbene-2-O-ß-D-glucoside, and the determination method is simple, sensitive, reliable and can be used in fast identifying the crude/processed root of P. multiflorum or as a method for overall quality control of root of P. multiflorum.

[HPLC specific chromatograms of Xingnaojing injection].

To establish and analyze the HPLC specific chromatograms of Xingnaojing injection manufactured by different factories. The separation was performed on a Thermo BDS Hypersil C18 column (4.6 mm×250 mm, 5 µm), with the mobile phase consisting of acetonitrile-0.02% formic acid aqueous solution for gradient elution. The flow rate was 1.0 mL•min⁻¹, and the column temperature was 35 ℃. The detection wavelength was set at 254 nm, and the sample size was 20 µL. Eleven chromatographic peaks were identified as characteristic peaks of HPLC specific chromatograms of Xingnaojing injection, after analyzing 29 batches of Xingnaojing injection samples. Compared with the reference substances, seven of them were identified as eucarvone, camphor, curcumenone, curcumenol, curdione, curzerenone and germacrone, respectively. HPLC specific chromatograms of Xingnaojing injection manufactured by three factories could be easily classified into three categories after investigation with computer-aided similarity evaluation system combined with principal component analysis. The established HPLC specific chromatograms provide a basis for scientific evaluation and effective control of the quality of Xingnaojing injection.

[Analysis of different dosage forms of Xiasangju granules on fingerprints and models using high performance liquid chromatography].

To establish the fingerprints of Xiasangju granules (with sugar and non-sugar forms) by HPLC, and provide reference for their identification and effective quality control. High performance liquid chromatography (HPLC) method was used to collect the fingerprints of 20 batches of non-sugar Xiasangju granules and 34 batches of sugar type Xiasangju granules. Their main different components were classified and screened by mode identification methods (principal component analysis, PCA, and orthogonal partial least squares discriminate analysis, OPLS-DA). The principal components were identified by comparing with reference standards. The fingerprints of Xiasangju granules (sugar type and non-sugar type) were established. PCA could not fully classify the two types of granules, while OPLS-DA could obviously classify these two different types of Xiasangju granules. Six components showed greatest difference between two types of granules, including salviaflaside, luteoloside and linarin. The developed mode identification method is helpful to control the overall quality of Xiasangju granules, and it provides an effective approach to quality evaluation.

[Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example].

This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC50 values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C18 Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 µg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.

Multi-Optimization of Ultrasonic-Assisted Enzymatic Extraction of Atratylodes macrocephala Polysaccharides and Antioxidants Using Response Surface Methodology and Desirability Function Approach.

Rhizoma Atractylodes macrocephala polysaccharides (RAMP) have been reported to have a variety of important biological activities. In this study, an ultrasonic-assisted enzymatic extraction (UAEE) was employed to obtain the highest extraction yield and strongest antioxidant activity of RAMP and optimized by a multi-response optimization process. A three-level four-factor Box-Behnken design (BBD) was performed as response surface methodology (RSM) with desirability function (DF) to attain the optimal extraction parameters. The DPPH scavenging percentage was used to represent the antioxidant ability of RAMP. The maximum D value (0.328), along with the maximum yield (59.92%) and DPPH scavenging percentage (13.28%) were achieved at 90.54 min, 57.99 °C, 1.95% cellulase and 225.29 W. These values were further validated and found to be in good agreement with the predicted values. Compared to the other extraction methods, both the yield and scavenging percentage of RAMP obtained by UAEE was favorable and the method appeared to be time-saving and of high efficiency. These results demostrated that UAEE is an appropriate and effective extraction technique. Moreover, RSM with DF approach has been proved to be adequate for the design and optimization of the extraction parameters for RAMP. This work has a wide range of implications for the design and operation of polysaccharide extraction processes.

[UPLC Fingerprint of Oldenlandia corymbosa].

OBJECTIVE: To establish the UPLC fingerprint of Oldenlandia corymbosa from different regions and to distinguish it from Oldenlandia diffusa. METHODS: UPLC procedure was performed on an ACQUITY UPLC BEH C18 (50 mm x 2. 1 mm, 1. 7 µm) column and eluted with a mobile phase consisted of methanol-l % acetic acid at a flow rate of 0. 2 mL/min. The column temperature was 30 °C . The detection wavelength was 254 nm. A matrix was constructed for similarity evaluation, cluster analysis and principle component analysis. RESULTS: The collected samples had a good similarity. A specificity fingerprint chromatogram was produced and 15 common peaks were designated. Samples were divided into four groups. CONCLUSION: It is a reliable and available method for specific identification of Oldenlandia corymbosa and for distinguishing Oldenlandia corynbosa and Oldenlandia diffusa.

[Research on biological detoxification of Chinese medicine containing aristolochic acid A by ten microorganisms].

This paper was aim to screen microorganisms with attenualed efficiency for Chinese medicine containing aristolochic acid A by liquid-state fermentation. Twelve Chinese medicine were detected by UPLC and aristolochic acid A was only founded in four species of Aristolochia, those were Caulis Aristolochiae Manshuriensis, Aristolochiae Radix, Aistolochia Contorta Bunge and Herba Aristolochiae Mollissima,but not in the others. With the four Chinese medicine containing aristolochic acid A as raw material, ten microorganisms were tested, and the content of aristolochic acid A was detected by UPLC. The results showed that one microorganism can decrease content of aristolochic acid A in all those four Chinese medicine.

[Study on UPLC fingerprint of Xiasangju Granules].

OBJECTIVE: To establish a UPLC fingerprint method of Xiasangju Granules. METHODS: UPLC analysis was performed on a Waters ACQUITY UPLC H-Class system and carried out at 30 °C on a Waters Column ACQUITY UPLC BEH C18 (2.1 mm x 50 mm, 1.7 µm). A binary gradient elution system was composed of acetonitrile (phase A)and 0.5% acetic acid solution (phase B). Detection was performed at the wavelength of 320 nm,the mobile flow rate was at 0.4 mL/min. A matrix including 16 variations (characteristic peaks area)and 12 samples was constructed for similarity evaluation, cluster analysis and principle component analysis. RESULTS: The results showed that the collected samples had a good similarity. A specificity fingerprint was produced and 16 characteristic peaks were designated. 12 samples were divided into 6 groups. CONCLUSION: It is a reliable, available and quick method for quality control of Xiasangju Granules.