Solid-phase microextraction of endogenous metabolites from intact tissue validated using a Biocrates standard reference method kit / 药物分析学报

Improved analytical methods for the metabolomic profiling of tissue samples are constantly needed.Currently,conventional sample preparation methods often involve tissue biopsy and/or homogenization,which disrupts the endogenous metabolome.In this study,solid-phase microextraction(SPME)fibers were used to monitor changes in endogenous compounds in homogenized and intact ovine lung tissue.Following SPME,a Biocrates AbsoluteIDQ assay was applied to make a downstream targeted metab-olomics analysis and confirm the advantages of in vivo SPME metabolomics.The AbsoluteIDQ kit enabled the targeted analysis of over 100 metabolites via solid-liquid extraction and SPME.Statistical analysis revealed significant differences between conventional liquid extractions from homogenized tissue and SPME results for both homogenized and intact tissue samples.In addition,principal component analysis revealed separated clustering among all the three sample groups,indicating changes in the metabolome due to tissue homogenization and the chosen sample preparation method.Furthermore,clear differences in free metabolites were observed when extractions were performed on the intact and homogenized tissue using identical SPME procedures.Specifically,a direct comparison showed that 47 statistically distinct metabolites were detected between the homogenized and intact lung tissue samples(P＜0.05)using mixed-mode SPME fibers.These changes were probably due to the disruptive homogenization of the tissue.This study's findings highlight both the importance of sample preparation in tissue-based metabolomics studies and SPME's unique ability to perform minimally invasive extractions without tissue biopsy or homogenization while providing broad metabolite coverage.

Determination of methylmercury in urine by direct mercury analyzer / 中华劳动卫生职业病杂志

Objective: To develop asolvent extraction-direct mercury analyzer method for determination of methylmercury in urine. Methods: After the urinehydrolyzesd by hydrobromic acid, methylmercury was extracted by tolueneand reverse-extracted from L-cysteine solution, it was then detectedbydirect mercuryanalyzer. Results: The linear range was 0.2-50.0 μg/L, and the related coefficient was 0.9999. The relative standard deviations (RSD) within the group were 5.04%-6.64%, and the RSD between the group were 5.65%-8.11 %. The average recovery efficiencies were 85.4%-95.5%. The detection limitation was 0.0482 μg/L and the quantification concentrations was 0.1607 μg/L. Conclusion: The method, which has low detection limit, high sensitivity, easy to operate, is stability for the determination of methylmercury in urine.

Effects of Peganum harmala L. Seed Extract on Culex pipiens (Diptera: Culicidae) / Efeitos do extrato de semente de Peganum harmala L. em Culex pipiens (Diptera: Culicidae)

Mosquito-borne diseases result in the loss of life and economy, primarily in subtropical and tropical countries, and the emerging resistance to insecticides is increasing this threat. Botanical insecticides are promising substitutes for synthetic insecticides. This study evaluated the larvicidal and growth index of Culex pipiens of four solvent extracts of Terminalia chebula, Aloe perryi, and Peganum harmala against Cx. pipiens. None of the 12 extracts exhibited larvicidal potential against third instars except the ethyl acetate extract of P. harmala. After 24 h of exposure, the LC50 value was 314.88 ppm, and the LC90 value was 464.19 ppm. At 320 ppm, the hatchability was 25.83%, and it resulted in 100% mortality. In addition, the eggs treated with the EtOAc extract of P. Harmala exhibited a long larval period compared with the control. The larval period continued for 12 days, and the pupal period took three days in the treatment groups. The growth index data also exhibited a decrease (0.00­7.53) in the treated groups compare with 8.5 in the control. The transformation of eggs into adults decreased with increasing concentrations. This paper is the first report on the development and growth index of Cx. pipiens potential using P. harmala seeds.

As doenças transmitidas por mosquitos resultam na perda de vidas e economia, principalmente em países subtropicais e tropicais, e a resistência emergente aos inseticidas está aumentando essa ameaça. Os inseticidas botânicos são substitutos promissores dos inseticidas sintéticos. Este estudo avaliou o índice larvicida e de crescimento de Culex pipiens de quatro extratos solventes de Terminalia chebula, Aloe perryi e Peganum harmala contra Cx. pipiens. Nenhum dos 12 extratos exibiu potencial larvicida contra o terceiro ínstar, exceto o extrato de acetato de etila de P. harmala. Após 24 horas de exposição, o valor LC50 era 314,88 ppm e o valor LC90 era 464,19 ppm. A 320 ppm, a eclodibilidade foi de 25,83% e resultou em 100% de mortalidade. Além disso, os ovos tratados com o extrato de EtOAc de P. harmala exibiram um longo período larval em comparação com o controle. O período larval continuou por 12 dias, e o período pupal durou três dias nos grupos de tratamento. Os dados do índice de crescimento também exibiram uma diminuição (0,00-7,53) nos grupos tratados em comparação com 8,5 no controle. A transformação de ovos em adultos diminuiu com o aumento das concentrações. Este artigo é o primeiro relatório sobre o índice de desenvolvimento e crescimento de Cx. potencial de pipiens usando sementes de P. harmala.

Effects of Peganum harmala L Seed Extract on Culex pipiens (Diptera: Culicidae) / Efeitos do extrato de semente de Peganum harmala L em Culex pipiens (Diptera: Culicidae)

Mosquito-borne diseases result in the loss of life and economy, primarily in subtropical and tropical countries, and the emerging resistance to insecticides is increasing this threat. Botanical insecticides are promising substitutes for synthetic insecticides. This study evaluated the larvicidal and growth index of Culex pipiens of four solvent extracts of Terminalia chebula, Aloe perryi, and Peganum harmala against Cx. pipiens. None of the 12 extracts exhibited larvicidal potential against third instars except the ethyl acetate extract of P. harmala. After 24 h of exposure, the LC50 value was 314.88 ppm, and the LC90 value was 464.19 ppm. At 320 ppm, the hatchability was 25.83%, and it resulted in 100% mortality. In addition, the eggs treated with the EtOAc extract of P. Harmala exhibited a long larval period compared with the control. The larval period continued for 12 days, and the pupal period took three days in the treatment groups. The growth index data also exhibited a decrease (0.00–7.53) in the treated groups compare with 8.5 in the control. The transformation of eggs into adults decreased with increasing concentrations. This paper is the first report on the development and growth index of Cx. pipiens potential using P. harmala seeds.

As doenças transmitidas por mosquitos resultam na perda de vidas e economia, principalmente em países subtropicais e tropicais, e a resistência emergente aos inseticidas está aumentando essa ameaça. Os inseticidas botânicos são substitutos promissores dos inseticidas sintéticos. Este estudo avaliou o índice larvicida e de crescimento de Culex pipiens de quatro extratos solventes de Terminalia chebula, Aloe perryi e Peganum harmala contra Cx. pipiens. Nenhum dos 12 extratos exibiu potencial larvicida contra o terceiro ínstar, exceto o extrato de acetato de etila de P. harmala. Após 24 horas de exposição, o valor LC50 era 314,88 ppm e o valor LC90 era 464,19 ppm. A 320 ppm, a eclodibilidade foi de 25,83% e resultou em 100% de mortalidade. Além disso, os ovos tratados com o extrato de EtOAc de P. harmala exibiram um longo período larval em comparação com o controle. O período larval continuou por 12 dias, e o período pupal durou três dias nos grupos de tratamento. Os dados do índice de crescimento também exibiram uma diminuição (0,00-7,53) nos grupos tratados em comparação com 8,5 no controle. A transformação de ovos em adultos diminuiu com o aumento das concentrações. Este artigo é o primeiro relatório sobre o índice de desenvolvimento e crescimento de Cx. potencial de pipiens usando sementes de P. harmala.

Effects of Peganum harmala L. Seed Extract on Culex pipiens (Diptera: Culicidae)

Abstract Mosquito-borne diseases result in the loss of life and economy, primarily in subtropical and tropical countries, and the emerging resistance to insecticides is increasing this threat. Botanical insecticides are promising substitutes for synthetic insecticides. This study evaluated the larvicidal and growth index of Culex pipiens of four solvent extracts of Terminalia chebula, Aloe perryi, and Peganum harmala against Cx. pipiens. None of the 12 extracts exhibited larvicidal potential against third instars except the ethyl acetate extract of P. harmala. After 24 h of exposure, the LC50 value was 314.88 ppm, and the LC90 value was 464.19 ppm. At 320 ppm, the hatchability was 25.83%, and it resulted in 100% mortality. In addition, the eggs treated with the EtOAc extract of P. Harmala exhibited a long larval period compared with the control. The larval period continued for 12 days, and the pupal period took three days in the treatment groups. The growth index data also exhibited a decrease (0.007.53) in the treated groups compare with 8.5 in the control. The transformation of eggs into adults decreased with increasing concentrations. This paper is the first report on the development and growth index of Cx. pipiens potential using P. harmala seeds.

Resumo As doenças transmitidas por mosquitos resultam na perda de vidas e economia, principalmente em países subtropicais e tropicais, e a resistência emergente aos inseticidas está aumentando essa ameaça. Os inseticidas botânicos são substitutos promissores dos inseticidas sintéticos. Este estudo avaliou o índice larvicida e de crescimento de Culex pipiens de quatro extratos solventes de Terminalia chebula, Aloe perryi e Peganum harmala contra Cx. pipiens. Nenhum dos 12 extratos exibiu potencial larvicida contra o terceiro ínstar, exceto o extrato de acetato de etila de P. harmala. Após 24 horas de exposição, o valor LC50 era 314,88 ppm e o valor LC90 era 464,19 ppm. A 320 ppm, a eclodibilidade foi de 25,83% e resultou em 100% de mortalidade. Além disso, os ovos tratados com o extrato de EtOAc de P. harmala exibiram um longo período larval em comparação com o controle. O período larval continuou por 12 dias, e o período pupal durou três dias nos grupos de tratamento. Os dados do índice de crescimento também exibiram uma diminuição (0,00-7,53) nos grupos tratados em comparação com 8,5 no controle. A transformação de ovos em adultos diminuiu com o aumento das concentrações. Este artigo é o primeiro relatório sobre o índice de desenvolvimento e crescimento de Cx. potencial de pipiens usando sementes de P. harmala.

Pharmaceutical amorphous solid dispersion: A review of manufacturing strategies

Amorphous solid dispersions (ASDs) are popular for enhancing the solubility and bioavailability of poorly water-soluble drugs. Various approaches have been employed to produce ASDs and novel techniques are emerging. This review provides an updated overview of manufacturing techniques for preparing ASDs. As physical stability is a critical quality attribute for ASD, the impact of formulation, equipment, and process variables, together with the downstream processing on physical stability of ASDs have been discussed. Selection strategies are proposed to identify suitable manufacturing methods, which may aid in the development of ASDs with satisfactory physical stability.

Polyphenolic extracts of walnut (Juglans regia) green husk containing juglone inhibit the growth of HL-60 cells and induce apoptosis

BACKGROUND: Juglone is a naphthoquinone currently obtained by chemical synthesis with biological activities including antitumor activity. Additionally, juglone is present in the green husk of walnut, which suggests evaluating the effect of GH extracts on carcinogenic cell lines. RESULTS: Walnut green husk ethanolic extract was obtained as 169.1 mg juglone/100 g Green Husk and antioxidant activity (ORAC) of 44,920 µmol Trolox Equivalent/100 g DW Green Husk. At 1 µM juglone in HL-60 cell culture, green husk extract showed an antiproliferative effect, but pure juglone did not; under these conditions, normal fibroblast cells were not affected. A dose-dependent effect on mitochondrial membrane potential loss was observed. Apoptosis of HL-60 was detected at 10 µM juglone. Despite high ORAC values, neither purified juglone nor the extract showed protective effects on HL-60 cells under oxidative conditions. CONCLUSIONS: Green husk extract generates an antiproliferative effect in HL-60 cells, which is related to an induction of the early stages of apoptosis and a loss of mitochondrial membrane potential. The normal cells were not affected when juglone is present at concentrations of 1 µM, while at higher concentrations, there is loss of viability of both cancerous and healthy cells.

Evaluation of Antifibrin, Antioxidant and Antimicrobial Activities of Betel Leaves (Piper betel L.)

Piper betel L. belongs to the family Piperaceae. It has been an important medicinal agent since ages in various traditional and folk systems of medicine. Leaves obtained from the local market were shade dried and powdered. Different solvents were used based on polarity to extract phytochemicals from this powder using a Soxhlet extractor and separated using rotary vacuum evaporator. Thin layer chromatography was run using different solvent systems in different ratios for identifying essential compounds of Piper betel and for standardizing the ratios at which better resolution of compounds taken place. Antimicrobial activities were tested on twelve bacterial and three fungal species. Also, anti fibrin activity was tested on erythrocytes by using the extracts obtained by the plant. The zone of inhibitions formed due to the anti microbial activity were measured and found that mixtures of ethyl acetate and ethanol were effective. The percentage of clot lysis was found to be appreciable for ethyl acetate extract of the Piper leaves.

Applications of new methods in extraction of flavonoids from Chinese materia medica / 中草药

Flavonoids are widely found in medicinal plants, which have important medical properties. Flavonoids were proved to have many pharmacological activities, such as anti-oxidation, antitumor, antimutation, anti-inflammatory, antibacterial and anti-aging. The extraction of flavonoids is the crucial link in their clinical applications. In recent years, many emerging Chinese medicine extraction methods have also been widely used in the extraction of flavonoids. This paper reviews the current application of new methods for flavonoid extraction, in order to provide references for the extraction, development and utilization of flavonoids. These new extraction methods include supercritical fluid extraction (SFE), ultrasonic assisted extraction (UAE), microwave assisted extraction (MAE), pressurized liquid extraction (PLE), pulsed electric field (PEF) assisted extraction, enzyme assisted extraction (EAE), green solvent extraction, steam explosion assisted extraction, dynamic high pressure microfluidization (DHPM) assisted extraction, etc.

Simultaneous Extraction and Separation of Oil and Azadirachtin from Seeds and Leaves of Azadirachta indica using Binary Solvent Extraction

Conventional extraction of oil and azadirachtin, a botanical insecticide, from Azadirachta indica involves defatting the seeds and leaves using hexane followed by azadirachtin extraction with a polar solvent. In order to simplify the process while maintaining the yield we explored a binary extraction approach using Soxhlet extraction device and hexane and ethanol as non-polar and polar solvents at various ratios and extraction times. The highest oil and azadirachtin yields were obtained at 6 h extraction time using a 50:50 solvent mixture for both neem leaves (44.7 wt%, 720 mg(Aza)/kg(leaves)) and seeds (53.5 wt%, 1045 mg(Aza)/kg(seeds)), respectively.

Ultrasound-assisted extraction produce better antibacterial and antioxidant activities of Senna siamea (Lam.) leaf extracts than solvent extraction

Aims@#Senna siamea has various medicinal functions but specific studies pertaining to the antioxidant and antibacterial potential that are related to ultrasound-assisted extraction from S. siamea have not been found to be reported yet. Therefore, this research was to determine antibacterial activities and antioxidant of S. siamea leaf extracts using solvent extraction and ultrasound-assisted extraction. @*Methodology and results@#Antibacterial activities were tested using the disc diffusion method and MIC and MBC values of seven bacterial strains. The ultrasound-assisted extraction extract had a higher yield, total phenolic content, antioxidant activities, and antibacterial activity than solvent extract. Interestingly, the strains of Staphylococcus sp., Vibrio parahaemolyticus, Escherichia coli, and Salmonella enteritidis were not inhibited by the solvent extracts, but were significantly (p < 0.05) inhibited by the ultrasound-assisted extraction extracts. Besides, the MIC and MBC values of extracts from ultrasound-assisted extraction were lower than the extracts from solvent extraction. @*Conclusion, significance and impact of study@#The results revealed that extracts from ultrasound-assisted extraction have higher efficiency to treat bacterial strains due to the efficiency of extraction method towards the recovery and solubility of extractable compounds. The results concluded that the extracted using ultrasound-assisted extraction can be used as active pharmaceutical components for the treatment, prevention, and control of pathogenic bacteria, including to be applied as food ingredients.

Ameliorative Potentials of Aqueous and Hydro-Ethanol Extracts of Citrullus colocynthis L Fruit on Hyperglycemia and Some Serum Biochemical Parameters on Alloxan-induced Diabetic Rats

Aims: This study was carry out to determine the dose effect of aqueous and hydro-ethanol C. colocynthis fruit extracts on some properties and serum biochemical parameters in alloxan-induced diabetic rats. Study Design: Random trial. Place and Duration of Study: University of Douala-Cameroon. Faculty of Science. Laboratory of Biochemistry, between April 2017 and June 2017. Methodology: Forty two albino male rats (Rattus norvegicus var. albinus), aged 2 to 3 months and weighing 150- 200 g were used in this study. Animals were randomly distributed into 7 experimental groups of six rats each. One group was used as healthy control and diabetes was induced in the 6 others groups by intraperitoneal injection with alloxan monohydrate into tail veins. Five diabetic groups received oral glibenclamide (3mg/kg), 50 and 100 mg/kg bw of aqueous or hydro-ethanol fruit extract of Citrullus colocynthis respectively The remaining group was assigned as diabetic control rats. Body weight and serum biochemical parameters (glucose, lipids, transaminases and creatinine) were recorded weekly during 3 weeks. Results: Diabetes induction of with alloxan significantly (p = .05) increases blood glucose, triglyceride, cholesterol, transaminases (AST, ALT) and creatinine blood level. Treatment of rats with hydro-ethanol extract (100 mg/kg.bw) steadily reduces (80.44%) glycemia but cause a significant increase of the liver relative mass, AST (138.38 IU/l) and ALT (152.35 IU/l) blood levels (p = .05). Administration of 50 mg/kg bw hydro-ethanol and aqueous extracts significantly reduce the glucose (22-46.44%), triglycerides, total cholesterol, transaminases and creatinine blood levels (72 to 85 %) (p = .05). Conclusion: The administration of glibenclamide (3mg/kg), 50 and 100 mg/kg bw aqueous extracts and 50 mg/kg bw hydro ethanol extracts significantly reversed the damage associated with Alloxan induced diabetes close to the normal. The dose of 100 mg/kg bw hydro ethanol extracts should be avoided because of it toxic effects.

Successive Solvent Extraction And Hptlc Of Stem Bark Of Asoka – Saraca Asoca (roxb.) De Wilde

Asoka – Saraca asoca (Roxb.) de Wilde, is a medium sized evergreen tree growing in tropical regions. It has been used for various medicinal purposes from the time immemorial. Ample citations about its usage can be elicited from Veda’s, Puranas and Samhitas. Owing to extensive use, lack of cultivation and irrational collection practices it became an endangered drug. It’s one among the five endangered plants listed by NMPB. This scarcity of drug in the market eventually led to adulteration. It is one of the severely adulterated drug next to Bala – Sida species. Various pharmacognostical and phytochemical techniques are evolved from time to time to check the adulteration. Due to the sophisticated methodologies used by medicinal plant dealers, these methods fail to check adulteration. Pharmacognostical analysis of sample drug and its powder microscopy serves as an effective method to check adulteration. But it won’t serve fruitful when the drug gets adulterated with exhausted samples. In such cases, effective marker compounds of the drug need to be analysed. This can be achieved by analysing successive solvent extractives of test drug and by HPTLC analysis. Here an attempt has been done to analyse the successive solvent extraction and HPTLC of stem bark of Asoka – Saraca asoca (Roxb.) de Wilde. as an effective methodology to ensure the purity. The successive solvent extraction revealed 1.78%, 0.4%, 13.63% & 27.69% of extractives respectively in petroleum ether, cyclohexane, acetone and methyl alcohol. The qualitative analysis also showed significance difference in the steroids, alkaloids, phenols and flavonoids in each solvent. The results are promising and suggestive of considering these experiments as an effective method to ensure the quality and purity of drug sample.

Determination of Astragaloside in Four Kinds of Chinese Patent Medicines by ASE-CAD Combined with Column Post Compensation Liquid Chromatography / 中国药学杂志

OBJECTIVE: To establish a method for determination of astragaloside in 4 Chinese patent medicines (BuZhong YiQi Pills, TongQiao BiYan Granules, XingNao ZaiZao Capsules, YuPing Feng Granules) by accelerate solvent extraction (ASE) combined with column post compensation liquid chromatography and charged aerosol detector (CAD) detection, and compare the result with that of the pharmacopoeia method. METHODS: The optimal extraction conditions were determined by the ASE test: water saturated n-butanol was used as solvent, ASE extraction temperature was 100℃, extraction time was 7 min and cycle time was 3. For the HPLC analysis, Thermo AQ-C18 column (2.1 mm × 100 mm, 3 μm) was employed, the mobile phase for the analysis pump was composed of acetonitrile-water (15:85), and the flow rate was 0.3 mL•min-1. Gradient program was as follows: 0 min, 15% A; 0 - 4 min, 15% - 60%A; 4 - 5 min 60% A. The mobile phase for the compensating pump was acetonitrile, and the flow rate was 0.3 mL•min-1. RESULTS: The linear range of the calibration curve of astragaloside was 26.62 - 665.5 μg•mL-1. The RSD of sample analysis was 1.0% - 2.1%. The average recoveries were 98.80% - 100.80%. The method is in good agreement with the Pharmacopoeia method. CONCLUSION: The method is rapid, accurate and reproducible. It can be used for the determination of astragaloside in 4 kinds of Chinese patent medicines.

Determination of 16 Kinds of Polycyclic Aromatic Hydrocarbons in Atmospheric Fine Particles by Accelerated Solvent Extraction Coupled with High Performance Liquid Chromatography / 分析化学

A method for determination of 16 kinds of polycyclic aromatic hydrocarbons ( PAHs ) in atmospheric fine particles ( PM2. 5) was developed based on accelerated solvent extraction-direct injection coupled with high performance liquid chromatography (HPLC). PM2. 5 was collected by glass fiber membrane filter and pretreated with acetonitrile by accelerated solvent extraction. The extract was separated by ZORBAX Eclipse PAH column with acetonitrile and water as mobile phase, and detected by ultraviolet and fluorescence detectors. The result showed that the 16 kinds of PAHs were well separated and there were good linear relationships ( r≥0. 9998) in the concentration range of 0. 025 -5. 00 μg / mL. The recoveries were from 78. 3% to 113. 2% . The relative standard deviations ranged from 0. 5% to 9. 5% . The detection limits were 0. 007 - 0. 062 ng / m3 . The method was simple, rapid, accurate and sensitive, and suitable for the simultaneous determination of 16 kinds of PAHS in PM2. 5.

Optimization of extraction method of total flavones in Lonicera japonica by orthogonal design / 国际中医中药杂志

Objective To optimize and establish the extraction of total flavones in Lonicera japonica. Methods The ultraviolet spectrophotometry was used to determinate the total flavonoids. The effect of extraction time, the amount of solvent and extraction times,pH were tested by the orthogonal design.ResultsThe total flavonoids showed a linear relationship,the optimal extraction condition of total flavones in Lonicera japonica was with 16 times volume of sample weight water for three times, each time was 2 h.Conclusions The method is simple and applicable to the extraction of total flavones in Lonicera japonica.

Rapid Determination of Chrysophanol and Aurantio-Obtusin in Cassia Obtusifolia L.by ASE-HPLC / 中国药师

Objective: To establish an accelerated solvent extraction(ASE)-HPLC method to determine chrysophanol and aurantio-obtusin in Cassia obtusifolia L.Methods: The optimal extraction conditions were defined by orthogonal tests using ASE.The method was carried out on an ACE Excel C18-PFP column (75 mm×2.1 mm,2.5 μm) with the mobile phase consisting of 0.1% phosphoric acid solution-acetonitrile with gradient elution.The column temperature was 40 ℃,the flow rate was 0.4 ml·min-1, and the detection wavelength was 284 nm. Results: The best process parameters of ASE were as follows:the extraction solvent was methanol, the extraction temperature was 120 ℃ and the static extraction duration was 5 minutes for three cycles.The ASE method needed only 1/9 of the time as the pharmacopoeia method,while the extraction efficiency of the ASE method was higher.The linear ranges of cassia obtusifolia L.and Chrysophanol were at 0.73~58.57 μg·ml-1(r=0.999 7) and 1.09~87.29 μg·ml-1(r=0.999 6).The average recoveries were 102.7%(RSD=0.8%) and 98.2%(RSD=1.5%).Conclusion: The method is simple, rapid and sensitive, which can be used for the rapid determination of aurantio obtusin and chrysophanol in Cassia obtusifolia L.

Simultaneous determination of four constituents in Zizyphi Spinosae Semen by ASE-CAD / 中草药

Objective: To establish an accelerated solvent extraction (ASE)-charged aerosol detector (CAD) method for simultaneous detection of jujuboside A, jujuboside B, spinosin, and betulinic acid in Zizyphi Spinosae Semen. Methods: The orthogonal design was applied to optimize the extraction parameters of the ASE system. The target compounds were detected by HPLC-CAD with the parameters as follow: Thermo Syncronis C18 (100 mm × 3 mm, 3 μm) column, a gradient elution program with acetonitrile-water as mobile phase at a flow rate of 0.5 mL/min, the column temperature was kept at 40℃. Detector was Corona Ultra CAD with 35℃ of nebulization temperature. Results: Optimization of the ASE parameters with orthogonal design greatly improved the extraction efficiency; All the target compounds could be simultaneously determined in a single run. Good linear relationships (0.998 3-0.999 6) and high relative recoveries were 98.46%-102.02%. Conclusion: The method is rapid, simple, accurate, and thus could be used for the quality control of Zizyphi Spinosae Semen.

Rapid determination of oleanolic acid and ursolic acid in eight kinds of Chinese materia medica by HPLC / 中草药

Objective: To establish a method of rapid analysis for simultaneous determination of oleanolic acid and ursolic acid in Chinese materia medica (CMM) of Verbena officinalis, Ligustrum lucidum, Prunella vulgaris, Hedyotis diffusa, Patrinia scabiosaefolia, Eriobotrya japonica, Crataegus pinnatifida, Chaenomeles Fructus papaya. Methods: The analyses of preparation were conducted by accelerated solvent extraction (ASE), methanol was used as solvent extraction, and the static extraction time was 6 min. Separation was carried out on Acclaim C30 Thermo column with acetonitrile and 0.2% acetic acid (85:15) as mobile phase, flow rate was 0.3 mL/min, and UV detection wavelength was set at 205 nm. Results: After 10 min sample extraction time, oleanolic acid and ursolic acid reached baseline separation, and no sample matrix was interfered, the linear correlation coefficient was over 0.999, the average recovery between 95.8% and 102.7%; Compared to Chinese Pharmacopoeia 2015, the determination of average mass fraction difference was 3.7% and 5.8%. Conclusion: This method is simple, fast, and suitable for the analysis of these drugs, and it can be used for content determination of oleanolic acid and ursolic acid for eight kinds of CMM.

Simultaneous Determination of 9 Common Inorganic Anions in Huanglian Shangqing Tables by Accelerat-ed Solvent Extraction-ion Chromatography / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 9 common inorganic anions in Huanglian shangqing tablet. METHODS:The accelerated solvent extraction-ion chromatography was adopted. Inorganic anions were deter-mined by Ion Pac AS11-HC anion exchange column,protected by Ion Pac AG11-HC column and eluted by hydroxide potassium so-lution(gradient elution)at the flow rate of 1.2 mL/min. The column temperature was set at 30 ℃,elution time was 18 min,and sample volume was 25 mL. RESULTS:The linear ranges of fluorinion,formate ion,nitrite ion,bromide ion,nitrate ion,sulfate ion,oxalate ion and phosphate ion were 0.1-5 mg/L(r=0.9990-0.9999). The limits of quantitation were 0.020,0.078,0.030, 0.058,0.052,0.068,0.084,0.064,0.074 mg/L,and the limits of detection were 0.005,0.024,0.008,0.017,0.015,0.022, 0.026,0.020,0.021 mg/L,respectively. RSDs of precision,stability and repeatability were all lower than 4.0%;recoveries were 80.00%-125.08%(RSD ranged 0.97%-2.47%). CONCLUSIONS:The method is simple,precise,stable and repeatable,and can be used for 9 common inorganic anions in Huanglian shangqing tablet.