The effect of photoperiodism on nutritional potency of Euglena sp. Indonesian strains

@#Introduction: Biomass microalgae functional materials, such as drugs and food supplements, have recently received much attention. Euglena sp. is a particularly appealing microalgae because of its simplicity of culture and capacity to produce a wide range of bioactive compounds. Moreover, it is one of the few microorganisms that produces carbohydrate, lipid, protein, ß-1,3-glucans, antioxidants, phytotoxins, wax esters, and polyunsaturated fatty acids that can be used to make nutraceuticals, pharmaceuticals, and cosmeceuticals. However, the potential utilisation of Euglena sp. for production of food supplements has been exploited only on a limited basis. Methods: This study was modified by adding protocatechuic acid and photoperiodism for 12:12; 14:10; 16:8; and full dark to affect the metabolite content of Euglenasp. Results: Results showed that the photoperiod had significant effect on lipid, chlorophyll-a, and carotenoid levels in the control treatment, with the highest levels as follows: 0.52±0.03 g/L, 1.20±0.01 x10-2 g/L, 0.30±0.02 x10-2 g/L; while the others were not significantly affected by the treatment, with the highest protein content at full dark 3.10±0.2 x10-2 g/L; chlorophyll-b at photoperiod 14:10 0.70±0.03 x10-2 g/L; paramylon at photoperiod 12:12 1.90±0.02 x10-1 g/L. The highest carbohydrates were found in control, with a level of 1.20±0.02 g/L. Conclusion: Photoperiodism is recommended to enhance productivity of protein, paramylon, and chlorophyll-b, while full light is recommended to enhance carbohydrate, lipid, chlorophyll-a, and carotenoid production in Euglena sp. to improve the quality of food nutrition.

HPLC quantitative analysis of protocatechuic acid contents in 11 Phellinus mushroom species collected in Thailand

Abstract Phellinus mushrooms have been traditionally used for various medicinal purposes. Protocatechuic acid, which was previously reported to be a component in some Phellinus mushrooms, has some pharmacological effects. This study aimed to validate a HPLC method for the quantitative analysis of the protocatechuic acid contents in the extracts from different Phellinus mushroom species collected in Thailand. HPLC was carried out using a C18 column and the gradient mobile phases of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Method validation was performed to assure the linearity, accuracy, precision, limit of detection and limit of quantitation of the analytical method. The linearity range of protocatechuic acid was 1 - 10 µg/ml. The average recovery was 104.16%. The method was shown to be precise with the RSD of repeatability and intermediate precision at less than 3%. The protocatechuic contents in 11 Phellinus mushrooms were in the range of less than 0.0099 - 0.4121 %w/w of the extract. The developed HPLC method was reliable and suitable for the quantitative analysis of protocatechuic acid content in Phellinus mushrooms.

Simultaneous determination of 10 components in Guanxinshutong capsules by quantitative analysis of multi-components by single marker / 药学学报

This study establishes a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of gallic acid, sodium danshensu, protocatechuic acid, protocatechuic aldehyde, vanillin, rosmarinic acid, salvianolic acid B, eugenol, cryptotanshinone and tanshinone IIA in Guanxinshutong capsules (Bambusae Concretio Silicea, Salvia miltiorrhiza, clove, borneol, Bambusae Concretio Silicea) by HPLC. Sample was loaded onto an Agilent C18 (ZORBAX Extend-RP C18, 250 mm × 4.6 mm, 5 µm) column and eluted with methanol-0.4% aqueous formic acid solution as a flow phase gradient, flow speed 1.0 mL·min-1, detection wavelength 280 nm, column temperature 35 ℃ and sample intake of 5 µL. Using protocatechuic acid as the internal reference, a relative correction factor was calculated and the durability was investigated, and the content of 10 components was calculated by QAMS and external standard method (ESM). The results show that the specificity, linear relationship, precision, repeatability, and stability of the 10 components were good. The average recovery was 98.20%-103.47% and RSD was 1.26%-2.84%. The relative positive factors and contents of the other nine components were calculated as gallic acid (0.759, 227.381), sodium tanshinol (3.630, 3.283), protocatechualdehyde (0.185, 0.150), vanillin (0.532, 65.213), rosmarinic acid (4.240, 1.035), salvianolic acid B (3.245, 18.204), eugenol (1.729, 9.265), cryptotanshinone (0.691, 1.449), and tanshinone ⅡA (0.702, 1.939). The results of QAMS were consistent with ESM analysis, and the relative error was between -3% and 3%. This method is stable and reliable, and can be used for the determination of 10 components in Guanxinshutong capsules.

Caffeic acid and protocatechuic acid modulate Nrf2 and inhibit Ehrlich ascites carcinomas in mice

Objective: To assess the nuclear factor-erythroid 2-related factor-2 (Nrf2) modulatory effect of caffeic acid and protocatechuic acid and determine the anti-tumor activity of these phenolic compounds against Ehrlich ascites carcinoma growth in mice. Methods: Antioxidant activity of protocatechuic acid and caffeic acid was assessed using ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Nrf2 activation potential of phenolic compounds was tested by quantitative realtime polymerase chain reaction, and luciferase complementation reporter assays. In vivo efficacy was tested using the Ehrlich ascites carcinoma model. Results: FRAP and DPPH radical scavenging assays showed that caffeic acid and protocatechuic acid were more potent compared with cinnamic acid and benzoic acid. Luciferase complementation reporter assays identified caffeic acid and protocatechuic acid as the activators of Nrf2. Both caffeic acid and protocatechuic acid upregulated the expression of Nrf2 target genes heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and glutamate-cysteine ligase modifier subunit (GCLM) and the activity of NAD(P)H:quinone oxidoreductase 1 (NQO1) when tested on HCT-116 cells using a cell-based assay system at 9 h. In addition, intraperitoneal administration of caffeic acid and protocatechuic acid to Ehrlich ascites carcinoma bearing mice suppressed tumor growth and angiogenesis. Conclusions: Caffeic acid and protocatechuic acid can modulate Nrf2 and inhibit Ehrlich ascites carcinoma cells.

Effects of Corchorus olitorius and protacatechuic acid on diabetic rat testis tissue / Efectos de Corchorus olitorius y ácido protocatéquico en el tejido testicular de rata diabética

SUMMARY: The aim of this study is to investigate the effects of Protocatechuic acid and Corchorus olitorius on streptozotocin (STZ) induced diabetic rat testis tissue. Randomly selected Wistar Albino rats were divided into five groups as; Diabetes Mellitus, Diabetes Mellitus treated with Corchorus Olitorus (STZ+CO), Diabetes Mellitus treated with Protacatechuic acid (STZ+PCA), Corchorus olitorius (CO), Protocatechuic acid (PCA) and Control. Diabetic model was generated by intraperitoneal injection of 60 mg/kg Streptozotosin. After 48 hours of the STZ injection, blood samples were collected from tail vein in order to measure blood glycose levels. Over 250 mg/dL accepted as diabetic subjets and fed with 250 mg/kg Corchorus olitorius or 20 mg/kg PCA by oral gavage for three weeks. At the end of the experiment, right testes were removed and fixed in 10 % neutral formaldehyde for paraffine embedding. Sections were stained by HE, Masson trichrome, PAS and TUNEL for microscopic evaluation. Control, PCA-only and Corchorus olitorius-only treated group testes tissues showed a normal tissue organization, when degeneration in seminiferous tubules, the vacuolization, seperations in spermatogenic cell series, outpouring of cell groups in the lumen, vesicular body formation, liquid accumulation in the interstitial region and edema were observed in STZ induced diabetic models and untreated groups. Besides, higher amount of TUNEL (+) stained cells were determined in STZ group. On the other hand, blood glucose level and number of TUNEL (+) stained cells were decreased as a result of PCA and Corchorus olitorius treatment. Because of the reduction of blood glucose level and apoptotic cell numbers, PCA and Corchorus olitorius decreace the complications of diabetes mellitus induced rat testis.

RESUMEN: El objetivo de este estudio fue investigar los efectos del ácido protocatéquico y Corchorus olitorius sobre el tejido testicular de rata diabética inducida por estreptozotocina (STZ). Las ratas Wistar Albino fueron seleccionadas al azar y se dividieron en cinco grupos; Diabetes Mellitus, Diabetes Mellitus tratada con Corchorus olitorius (STZ + CO), Diabetes Mellitus tratada con ácido protocatéquico (STZ + PCA), Corchorus olitorius (CO), ácido protocatéquico (PCA) y Control. El modelo diabético se generó por inyección intraperitoneal de 60 mg/kg de estreptozotosina. Después de 48 horas de la inyección de STZ, se recogieron muestras de sangre de la vena de la cola para medir los niveles de glucosa. Niveles mayores a 250 mg/dL fueron considerados como especímenes diabéticos y alimentados con Corchorus olitorius de 250 mg/kg o PCA de 20 mg/kg por sonda oral durante tres semanas. Al final del experimento, se extirparon los testículos derechos y se fijaron en formaldehído neutro al 10 % para la inclusión en parafina. Las secciones se tiñeron con HE, tricromo de Masson, PAS y TUNEL para evaluación microscópica. Los tejidos de los testículos de los grupos control, tratados solo con PCA y con Corchorus olitorius mostraron una organización tisular normal. En cambio en modelos diabéticos inducidos por STZ y grupos no tratados se observó degeneración en los túbulos seminíferos, vacuolización, separaciones en series de células espermatogénicas, efusión de grupos celulares en la luz, formación del cuerpo vesicular, acumulación de líquido en la región intersticial y edema. Además, se determinó una mayor cantidad de células teñidas con TUNEL (+) en el grupo STZ. Por otro lado, el nivel de glucosa en sangre y el número de células teñidas con TUNEL (+) disminuyeron como resultado del tratamiento con PCA y Corchorus olitorius. Debido a la reducción del nivel de glucosa en sangre y el número de células apoptóticas, se observó que PCA y Corchorus olitorius disminuyen las complicaciones de los testículos de rata inducidos por diabetes mellitus.

UPLC Fingerprint Analysis of Acanthopanacix Cortex from Different Origins / 中国实验方剂学杂志

Objective::To established fingerprint of Acanthopanacix Cortex by UPLC method, in order to provide reference for quality control and evaluation. Method::UPLC method was performed on Waters BEH C18 (2.1 mm×100 mm, 1.7 μm), with acetonitrile-0.1% glacial acetic acid as the mobile phase for gradient elution.The detection wavelength was 282 nm, the flow rate was 0.3 mL·min-1, the column temperature was 25 ℃, and the injection volume was 2 μL.With syringin as reference substance, the fingerprint of 20 batches Acanthopanacix Cortex were analyzed under the same chromatographic conditions.The Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Media (version 2012) was used to analyze the similarity of 20 batches of Acanthopanacix Cortex, and the SPSS 21.0 was applied for cluster analysis. Result::The UPLC fingerprint of the Acanthopanacix Cortex was established.The similarity results showed that the 7 batches of the 20 batches of Acanthopanacix Cortex was less than 0.800, and the remaining medicinal materials were similar within the range from 0.800 to 0.924.Besides, 12 common fingerprint peaks were calibrated and 4 components were identified, namely protocatechuic acid (peak 1), chlorogenic acid (peak 3), syringin (peak 4), and 4-methoxysalicylaldehyde (peak 12). The clustering results showed that the 20 batches of Acanthopanacix Cortex were divided into four groups.Among these batches, S1, S3, S9, S13 and S20 were clustered into one category, S11 was a category, S14 was a category, and the remaining samples belonged to a category. Conclusion::With a good precision, repeatability and stability, short analysis time as well as superior specificity, the method will provide a scientific basis to evaluate and control the quality of Acanthopanacix Cortex.

Protocatechuic acid alleviates degeneration of chondrocytes / 中国组织工程研究

BACKGROUND: Preliminary studies have shown that numerous antioxidants exhibit antiarthritic effects due to their inhibition on inflammatory factors. With the antioxidant and anti-inflammatory abilities whether protocatechuic acid is effective in the treatment of osteoarthritis has never been reported. OBJECTIVE: To investigate the biological effect of protocatechuic acid on interleukin-1β induced chondrocyte including phenotype and cellular metabolism in vitro, thereby providing a potential agent in osteoarthritis treatment, METHODS: The chondrocytes of neonatal rat femur were collected, intervened by 10 mg/L interleukin-1β to establish the degenerative model, and treated by 10, 30 and 50 mg/L protocatechuic acid. The study was approved by the Laboratory Animal Ethical Committee of Guangxi Medical University in October 2017, with the approval No. 201710008. RESULTS AND CONCLUSION: Protocatechuic acid effectively promoted chondrocyte growth in the range of 10-50 mg/L, while the dose of 30 mg/L was the strongest. Protocatechuic acid also enhanced the synthesis of the extracellular matrix and the mRNA expression of aggrecan, collagen II and Sox9, and downregulating the expression of the matrix metalloproteinase 13 (a marker of inflammatory factor). To conclude, protocatechuic acid exerts a positive effect on the proliferation and phenotypic maintenance of articular chondrocytes, providing reference for its use in the treatment of osteoarthritis and repair of degenerative articular cartilage in vivo.

Identification and comprehensive quality evaluation of Cynomorium songaricum in different producing areas based on OPLS combined with entropy weight TOPSIS method / 中草药

Objective: To identify and comprehensively evaluate Cynomorium songaricum from different producing areas in order to provide reference for the quality evaluation of C. songaricum and the determination of the suitability of the origin. Methods: A total of 40 samples from five provinces (regions) were collected to measure the content of gallic acid, protocatechuic acid, catechins, total polysaccharides, total flavonoids, Na, K, Ca, Mg, Fe, Zn, Mn, Co, Sr, Ni, Ag, Ba, Ti, Cu, Pb, Cr, Cd, As, and Hg. The data reflecting the quality of C. songaricum were analyzed by orthogonal partial least squares discriminant analysis (OPLS-DA) and entropy weight TOPSIS analysis. Results: The contents of Mn, Zn, Co, catechin, Pb, Cr, Ca, Ti, total flavonoids, protocatechuic acid, Mg and Cu in C. songaricum are important for distinguishing different producing areas. The quality of C. songaricum in Inner Mongolia was the best in all provinces (regions), followed by Gansu, Ningxia, Xinjiang, and Qinghai Provinces. Conclusion: The results of OPLS-DA combined with entropy weight TOPSIS analysis are reasonable, objective and effective, and can be applied to the comprehensive evaluation of multiple indicators of C. songaricum.

Chemical constituents of Dendrobium hercoglossum / 中草药

Objective: To study the chemical constituents of Dendrobium hercoglossum. Methods: The compounds were isolated and purified by column chromatography and preparative HPLC, and their chemical structures were identified on the basis of spectral analysis. Results: Ten compounds were isolated and identified as 3,4,α-trihydroxy-5,3'-dimethoxybibenzyl (1), 4,α-dihydroxy-3,5,3'- trimethoxybibenzyl (2), 4,5-dihydroxy-3,3',α-trimethoxybibenzyl (3), 4,3'-dihydroxy-3,5-dimethoxybibenzyl (4), 4,4'-dihydroxy- 3,5,3'-trimethoxybibenzyl (5), N-trans-cinnamoyltyramine (6), protocatechuic acid (7), vanillyl alcohol (8), hexadecanoic acid 2,3-dihydroxypropyl ester (9) and syringaresinol (10). Conclusion: Compound 1 is a new compound, named dendhercoine A, compounds 2-10 are isolated from this plant for the first time.

Simultaneous determination of thirteen components in Ainsliaea fragrans by UHPLC-MS/MS / 中草药

Objective: To establish an UHPLC-MS/MS method for rapid and simultaneous determination of the content of 13 components in Ainsliaea fragrans. Methods: The UHPLC-MS/MS method was performed on UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm) with acetonitrile-water (containing 0.1% formic acid and 5 mmol ammonium formate) as mobile phase for gradient elution. Flow rate was 0.25 mL/min and column temperature was 40 ℃. A triple quadrupole mass spectrometer equipped with electrospray ionization source (ESI) was applied in negative ion mode with the following parameters: ion spray, -4 500 V, ion source temperature, 500 ℃, curtain gas, 344.5 kPa, nubulizer (GS1), 344.5 kPa, heater gas (GS2), 206.7 kPa, and multiple reaction monitoring (MRM) was performed for quantitative analysis of these compounds. Results: Under the optimized MS/MS condition, the linearity ranges of protocatechuic acid, caffeic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, luteolin, luteoloside and apigenin were 1.099-274.800 ng/mL, 1.006-100.600 ng/mL, 1.080-1 080.000 ng/mL, 83.12-8 312.00 ng/mL, 19.92-996.00 ng/mL, 1.076-269.000 ng/mL, 5.555-555.500 ng/mL, 4.420-4 420.000 ng/mL, 44.76-17 904.00 ng/mL, 48.00-9 600.00 ng/mL, 2.108-210.800 ng/mL, 4.136-413.600 ng/mL, 1.070-107.000 ng/mL (r ≥ 0.9958), respectively, with good linearity. The average recovery rate of thirteen compounds in the samples were in the range of 96.54%-99.75%, and the RSD range was from 0.48% to 0.96%. The RSD values of precision, stability and repeatability test were all less than 3.90%. Conclusion: The method had good repeatability, high specificity, stability and controllability, and could be used for quality control of A. fragrans and its preparations.

Content Determination of Nine Constituents in Different Medicinal Parts of Pimpinella thellungiana / 中国实验方剂学杂志

Objective:To develop a method to quantify nine constituents in different medicinal parts of Pimpinella thellungiana, in order to compare the content difference of the nine constituents, namely protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C. Method:The analysis was performed on an Agilent TC-C18 column (4.6 mm×250 mm,5 μm) with acetonitrile and mixed acid solution (0.1% phosphoric acid-0.1% glacial acetic acid) as mobile phase for gradient elution. The handover detection wavelengths were at 265 and 325 nm. The column temperature was 20℃, and the flow rate was 1.0 mL·min-1. The experiment data was analyzed using the software of Markerlynx XS. Result:The nine constituents of protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid, luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C had a good degree of separation and a good linearity in their respective linear ranges(r>0.999 8). The average recoveries ranged from 99.11% to 100.76%,and the RSD ranged from 0.9% to 2.0% 。The results showed that the contents of the nine constituents had significant differences in different medicinal parts of P. thellungiana. The average contents of the nine constituents were the highest in leaves,which was followed by stem,and the lowest was in root. Conclusion:The study could provide evidence for the quality control,clinical application,and scientific resources utilization of P. thellungiana.

Phenolic Constituents from Caesalpinia decapetala / 中国药学杂志

OBJECTIVE: To study the phenolic constituents from Caesalpinia decapetala (Roth) Alston. METHODS: The ethanol crude extract of C. decapetala was fractionalized by using petroleum ether and chloroform. The chloroform part was isolated by a series of chromatography methods, and the structures of purified compounds were elucidated by their physicochemical properties and spectroscopic data. RESULTS: Eleven phenolic compounds were isolated and identified as methyl 2,3,5-trihydroxybenzoate (1), protocatechuic acid methyl ester (2), N-trans-feruloyl tyramine (3), trichostachine (4), cinnamylpiperidine (5), gallic acid (6), methyl 3,4,5-trihydroxybenzoate (7), ethyl 3,4,5-trihydroxybenzoate (8), resveratrol(9), 3,4,3',5'-tetrahydroxydistyrene (10), and protosappanin A (11). CONCLUSION: Compounds 1-5 are isolated from this plant for the first time.

Chemical constituents from fruits of Tribulus terrester / 中草药

Objective: To study the chemical constituents from the fruits of Tribulus terrester. Methods: The constituents were isolated and purified by silica gel column chromatography, Sephadex LH-20 column chromatography, preparative HPLC and TLC methods. The structures were identified by HR-ESI-MS and spectral analysis methods. Results: Fourteen compounds were isolated from the fruits of Tribulus terrester and the structures were identified as N-p-feruloyltyramine (1), N-trans-p-coumaroyl-3-O- methyldopamine (2), methyl ferulic acid (3), caffeic acid methyl ester (4), protocatechuic acid methyl ester (5), trifilines A (6), 5β-spirost-25(27)-en-3β-ol-12-one 3-O-{β-D-glucopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranoside} (7), tribulosaponin B (8), isoterrestrosin B (9), 25(R)-spirostane-3,5-diene-12-one (10), 25 (R)-spirostane-24β-ol-4-ene-3,12-dione (11), 26-O-β-D-glucopyranosyl-(25S)-5α-furostane-20 (22)-en-12-one-3β,26-diol-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl- (1→4)]-β-D-galactopyranoside (12), terrestrinone A1 (13), and terrestrinone A2 (14). Conclusion: Compounds 1-7 are obtained from the Zygophllyaceae for the first time.

Chemical constituents from Tibetan herbal medicines Biebersteinia heterostemon / 中草药

Objective To study the chemical components from aerial part of Tibetan herbal medicines Biebersteinia heterostemon. Methods The chemical constituents were isolated and purified by silica gel, Sephadex LH-20 chromatography and preparative liquid chromatography. Their structures were identified by physicochemical properties and spectral analysis. Results Thirteen compounds were isolated from B. heterostemon and their structures were identified as umbelliferone (1), 5,7,3’-trihydroxy-8,4’,5’- trimethoxyflavone (2), luteolin (3), quercetin (4), protocatechuic acid methyl ester (5), apigenin (6), alternariol (7), luteolin-7-O-β-D- glucoside (8), quercetin-3-O-β-D-glucoside (9), (+)-dehydrovomifoliol (10), β-sitosterol (11), 4’-methoxytricetin (12), and 3’,4’,5,8- tetrahydroxyflavanone-7-O-β-glucopyranoside (13). Conclusion Compounds 3, 5-7, 10, 11-13 are isolated from B. heterostemon for the first time. Compounds 5, 7, 10, 12, 13 are isolated from the genus Biebersteinia for the first time.

Isolation and identification of chemical constituents from Sabia parviflora (II) / 中草药

Objective: To study the chemical constituents from Sabia parviflora. Methods: Various column chromatographic techniques were used to separate and purify the chemical constituents which structures were elucidated by spectral analysis. Results: Fifteen compounds were isolated and identified as 1,3,5-trimethoxybenzene (1), vanillic aldehyde 4-O-β-D-glucopyranoside (2), 2,6-dimethoxy-4-hydroxybenzyl alcohol 1-O-β-D-glucopyranoside (3), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo [3,2,1]-oct-8-yl)-3-methyl-2Z,4E-penta-dienoic acid (4), protocatechuic acid (5), 4-quinolinone-2-caboxylic acid (6), 3,4,5-trimethoxy benzoic acid (7), pyrocatechol (8), syringic acid-4-O-β-D-glucopyranoside (9), (-)-(7α,8S)-erythro-1-C-syringylglycerol 4-O-β-D- glucopyranoside (10), 3,4,5-trimethoxy benzoic acid (11), 1,2,4-trihydroxybenzene (12), ferulic acid (13), vanillic acid (14), and p-hydroxybenzoic acid (15). Conclusion: Compounds 1-11 are isolated from the genus of Sabia for the first time, and compounds 12-15 are isolated from this plant for the first time.

Chemical constituents of Phyllanthus emblica and its anti-inflammation activities / 中草药

Objective: To study the chemical constituents and its anti-inflammtory activity effect of Phyllanthus emblica. Methods: The chemical constituents of P. emblica were isolated and purified by silica gel column chromatography, ODS column chromatography, Sephadex LH-20 column chromatography, semi-preparative high-performance liquid chromatography and recrystallization method. Through their spectra data, physical and chemical properties analysis, the structures of those compounds with high content were identified. LPS-induced RAW264.7 inflammatory cell model was established to evaluate the effect of compounds in P. emblica on proinflammatory factors (NO, IL-6, TNF-α, and MCP-1) of RAW264.7 inflammatory cells. Results: Totally, 14 compounds were isolated from P. emblica. and idetified as isovanillic acid (1), trans-cinnamic acid (2), p-hydroxybenzaldehyde (3), coniferyl aldehyde (4), quercetin (5), kaempferol-3-O-α-L-rhamnose (6), naringenin (7), 2-hydroxy-3-methyl phenylpropiolate (8), hydroquinone (9), myricetin (10), 2-furoic acid (11), methyl gallate (12), protocatechuic acid (13), gallic acid (14). The experiment of anti-inflammatroy effects showed that those compounds had different inhibitory effects on the production of inflammatory factors NO, IL-6, TNF-α and MCP-1. Conclusion: Compounds 1, 3, 4, 8-11 and 13 are isolated from P emblica for the first time. The anti-inflammatory effect of P. emblica is related to its phenolic acids.

Establishment of HPLC fingerprints of Kangfuxin Liquid and determintion of six constituents / 中成药

AIM To establish the HPLC fingerprints of Kangfuxin Liquid (extract of Periplaneta americana L.) and to determine the contents of six constituents.METHODS The analysis of this drug was performed on a TOSOH TSK-GEL ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.07％ acetic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.RESULTS There were twenty-four common peaks in the fingerprints of ten batches of samples (Ⅰ-Ⅹ) with the similarities of 0.932-0.993 (except for sample Ⅰ).Uracil,hypoxanthine,xanthine,inosine,protocatechuic acid and Cyclo (Gly-Tyr) showed good linear relationships within the ranges of 3.460-173.0,3.960-198.0,3.596-179.8,1.338-66.9,3.672-183.6 and 3.552-177.6 μg/mL,whose average recoveries (RSDS) were99.8％ (2.65％),98.0％ (2.55％),99.7％ (1.59％),100.7％ (2.80％),102.0％ (2.09％) and 99.6％ (1.88％),respectively.CONCLUSION This accurate,stable and simple method can be used for the quality control of Kangfuxin Liquid.

HPLC fingerprint of Homalomena occulta / 中草药

Objective: To establish an HPLC method for the fingerprint analysis of Homalomena occulta and evaluate the quality though similarity calculation. Methods: Waters e2695 was applied to analyze ten batches of H. occulta. Phenomenex Gemini 110A C18 column was performed (250 mm × 4.6 mm, 5 μm) with acetonitrile and 0.01% phosphoric acid in agradient mode at a flow rate of 1.0 mL/min at 290 nm and the separation was performed at 30 ℃. The similarity was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2004A. Results: The HPLC characteristic fingerprint of H. occulta with 11 commons peaks, and one of them were identified by comparing with reference subslances. The values of similarity was higher than 0.887. The HPLC method showed better results of stability, precision, and repeatability. Conclusion: It is the first time to establish the HPLC fingerprint of H. occulta. Those described can not only provides an identification method for fingerprint, but also provides the foundation of quality control.

Simultaneous determination of protocatechuic acid, salidroside, and chlorogenic acid in sargentodoxa cuneata by HPLC / 国际中医中药杂志

Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.

Simultaneous Determination of Six Active Constituents in Jizhi Syrups by HPLC with Wavelength Switc-hing / 中国药师

Objective:To establish an HPLC method for the simultaneous determination of six active constituents ( ferulic acid, chlorogenic acid,ephedrine hydrochloride,protocatechuic acid,protocatechualdehyde and naringin) in Jizhi syrups. Methods:A Zorbax XDB-C18(4.6 mm×250 mm,5 μm)column was used. The mobile phase consisted of acetonitrile (A)-0.9% acetic acid (B) with gradient elution at the flow rate of 1. 0 ml·min-1 . The detection wavelength was changed as follows:257 nm for 0-22. 0 min, 326 nm for 22. 0-30. 0 min, 320 nm for 30. 0-52. 0 min, 210 nm for 52. 0-58. 0 min and 283 nm for 58. 0-60. 0 min. The column temperature was 30 ℃ and the injection volume was 10 μl. Results:The separation of the six active constituents was good. The linear range ( r>0. 9990) was 2. 817-112. 670, 2. 342-93. 670, 0. 710-28. 415, 0. 776-31. 035, 0. 694-27. 755 and 1. 279-51. 175 ng for ferulic acid, chlorogenic acid, ephedrine hydrochloride, protocatechuic acid, protocatechualdehyde and naringin, respectively. The average recover-ies varied from 99.3% to 99.8%(RSD varied from 0.18% to 0.28%). Conclusion: The method is rapid with high sensitivity, promising accuracy and good specificity, which can provide scientific basis for the quality control of Jizhi syrups.