Structured Review

Shimadzu Corporation preparative hplc
<t>HPLC</t> profile of <t>meilingmycins</t> and structure comparison between meilingmycins and avermectin. (A) HPLC profile of meilingmycins. (B) Structures of meilingmycins and avermectin A1a. The carbons of meilingmycins are numbered. The five meilingmycins are different
Preparative Hplc, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 94/100, based on 298 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 298 article reviews
Price from $9.99 to $1999.99
preparative hplc - by Bioz Stars, 2020-11
94/100 stars

Images

1) Product Images from "Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †"

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02262-09

HPLC profile of meilingmycins and structure comparison between meilingmycins and avermectin. (A) HPLC profile of meilingmycins. (B) Structures of meilingmycins and avermectin A1a. The carbons of meilingmycins are numbered. The five meilingmycins are different
Figure Legend Snippet: HPLC profile of meilingmycins and structure comparison between meilingmycins and avermectin. (A) HPLC profile of meilingmycins. (B) Structures of meilingmycins and avermectin A1a. The carbons of meilingmycins are numbered. The five meilingmycins are different

Techniques Used: High Performance Liquid Chromatography

2) Product Images from "Bioactive Constituents of Lamium album L. as Inhibitors of Cytokine Secretion in Human Neutrophils"

Article Title: Bioactive Constituents of Lamium album L. as Inhibitors of Cytokine Secretion in Human Neutrophils

Journal: Molecules

doi: 10.3390/molecules23112770

HPLC-chromatograms of isolated compounds recorded at λ = 240 nm. HPLC conditions: Zorbax SB-C 18 (150 × 2.1 mm, 1.9 μm), mobile phase: A. 0.1% HCOOH/H 2 O; linear gradient 0–60 min, 5% to 60% B. 0.1% HCOOH/MeCN.
Figure Legend Snippet: HPLC-chromatograms of isolated compounds recorded at λ = 240 nm. HPLC conditions: Zorbax SB-C 18 (150 × 2.1 mm, 1.9 μm), mobile phase: A. 0.1% HCOOH/H 2 O; linear gradient 0–60 min, 5% to 60% B. 0.1% HCOOH/MeCN.

Techniques Used: High Performance Liquid Chromatography, Isolation

3) Product Images from "C-Glycosylated flavonoids from black gram husk: Protection against DNA and erythrocytes from oxidative damage and their cytotoxic effect on HeLa cells"

Article Title: C-Glycosylated flavonoids from black gram husk: Protection against DNA and erythrocytes from oxidative damage and their cytotoxic effect on HeLa cells

Journal: Toxicology Reports

doi: 10.1016/j.toxrep.2016.08.006

HPLC chromatogram of black gram milled by-product fractions showing vitexin (1) and isovitexin (2). (A) Whole gram; (B) Dhal; (C) Germ; (D) Aleurone; (E) Plumule; (F) Husk.
Figure Legend Snippet: HPLC chromatogram of black gram milled by-product fractions showing vitexin (1) and isovitexin (2). (A) Whole gram; (B) Dhal; (C) Germ; (D) Aleurone; (E) Plumule; (F) Husk.

Techniques Used: High Performance Liquid Chromatography

RP-HPLC chromatograms of flavone C -glycosides purified from husk extract. (A) vitexin (B) isovitexin.
Figure Legend Snippet: RP-HPLC chromatograms of flavone C -glycosides purified from husk extract. (A) vitexin (B) isovitexin.

Techniques Used: High Performance Liquid Chromatography, Purification

Related Articles

High Performance Liquid Chromatography:

Article Title: Dolichocarpols A-F, Unprecedented Macrocyclic Humulene-Type Sesquiterpenoids from Anaxagorea dolichocarpa
Article Snippet: .. Fraction H-2 (95.0 mg) was subjected to preparative HPLC with the following gradient elution: solvent A = H2 O; solvent B = MeOH; elution system = 0–40 min (20–70% B); 40–50 min (70% B); and 50–65 min (70–100% B), yielding compound 5 (2.0 mg). .. Fraction H-4 (85.8 mg) was also subjected to preparative HPLC using gradient elution with H2 O–MeOH 5 to 100% of MeOH in 75 min to afford compound 1 (1.0 mg).

Article Title: Methyl Salicylate Glycosides in Some Italian Varietal Wines
Article Snippet: .. Flash Chromatography with Isolute ENV+ and Preparative HPLC for the Isolation of the Single Glycosides A Shimadzu SCL-10 AVP preparative HPLC system with a Shimadzu SPD-10 AVP UV-VIS detector, 8A pumps, and Software Class VP (Shimadzu Corp., Kyoto, Japan) were used for chromatographic separation. .. For flash chromatography, 30 g of ENV+ resin (Biotage, Uppsala, Sweden) were conditioned with 300 mL of methanol and 500 mL of milliQ water.

Article Title: C-Glycosylated flavonoids from black gram husk: Protection against DNA and erythrocytes from oxidative damage and their cytotoxic effect on HeLa cells
Article Snippet: .. 2.5 Purification of vitexin and isovitexin of aqueous ethanol fraction by preparative HPLC The chromatographic separation was performed according to the method described in Section , on a Shimadzu Prep LC8A Preparative Chromatography system equipped with SCL-10AVP system controller (Shimadzu). .. The preparative HPLC was performed on a Varian, Pursuit XRs 10 C18 preparative column (250 mm × 21.2 mm).

Article Title: Bioactive Constituents of Lamium album L. as Inhibitors of Cytokine Secretion in Human Neutrophils
Article Snippet: .. Fraction DE6 (640 mg) was subjected to preparative HPLC (Shimadzu LC10vp, Japan, Zorbax SB-C18 —5 mm, 150 mm × 21.2 mm, Agilent (Agilent, Santa Clara, CA, USA), 280 nm, flow 20 mL·min−1 , mobile phase: 0.1% HCOOH in water (A) and 0.1% HCOOH in acetonitrile (B); elution program: 0% B–40% B (0–60 min), 40% B–50% B (60–65 min). ..

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: .. Together with fractions 9 and 10, five different meilingmycins were purified by preparative HPLC for the second time with the same condition to yield meilingmycins D (4.7 mg), E (2.9 mg), A (12.4 mg), B (3.9 mg), and C (2.1 mg). .. Nuclear magnetic resonance (NMR) spectra were recorded with a Bruker AM 500 NMR spectrometer in CDCl3 at 25°C, using the solvent as internal reference downfield of tetramethylsilane (TMS) at 0 ppm.

Article Title: Genotyping-Guided Discovery of Persiamycin A From Sponge-Associated Halophilic Streptomonospora sp. PA3
Article Snippet: .. Fractions containing a brown and a red compound were concentrated and subjected to preparative HPLC, LC-20 AP with a diode array detector SPD-M20A (Shimadzu) with a reverse-phase column (Phenomenex, Kinetex 5 μm, C18 250 × 10 mm). ..

Article Title: Antiobesity Effects of Gentiana lutea Extract on 3T3-L1 Preadipocytes and a High-Fat Diet-Induced Mouse Model
Article Snippet: .. Column chromatography (CC) was performed using Silica gel 60 (70~230 mesh, Merck, Darmstadt, Germany), ODS-A (12 nm S-7 μm, YMC GEL, Tokyo, Japan), and Preparative HPLC was performed on LC-8A (Shimadzu, Kyoto, Japan). .. Mouse 3T3-L1 preadipocytes were incubated in Dulbecco’s modified Eagle’s medium supplemented with streptomycin (100 µg/mL), penicillin (100 U/mL), and 10% fetal bovine serum.

Article Title: Activation of a Silent Fungal Polyketide Biosynthesis Pathway through Regulatory Cross Talk with a Cryptic Nonribosomal Peptide Synthetase Gene Cluster ▿Activation of a Silent Fungal Polyketide Biosynthesis Pathway through Regulatory Cross Talk with a Cryptic Nonribosomal Peptide Synthetase Gene Cluster ▿ †
Article Snippet: .. Preparative HPLC was performed using a Shimadzu HPLC system (LC-8A) with PDA and a C18 column (250 by 20 mm; Eurospher 100-5). ..

Flow Cytometry:

Article Title: Bioactive Constituents of Lamium album L. as Inhibitors of Cytokine Secretion in Human Neutrophils
Article Snippet: .. Fraction DE6 (640 mg) was subjected to preparative HPLC (Shimadzu LC10vp, Japan, Zorbax SB-C18 —5 mm, 150 mm × 21.2 mm, Agilent (Agilent, Santa Clara, CA, USA), 280 nm, flow 20 mL·min−1 , mobile phase: 0.1% HCOOH in water (A) and 0.1% HCOOH in acetonitrile (B); elution program: 0% B–40% B (0–60 min), 40% B–50% B (60–65 min). ..

Chromatography:

Article Title: Methyl Salicylate Glycosides in Some Italian Varietal Wines
Article Snippet: .. Flash Chromatography with Isolute ENV+ and Preparative HPLC for the Isolation of the Single Glycosides A Shimadzu SCL-10 AVP preparative HPLC system with a Shimadzu SPD-10 AVP UV-VIS detector, 8A pumps, and Software Class VP (Shimadzu Corp., Kyoto, Japan) were used for chromatographic separation. .. For flash chromatography, 30 g of ENV+ resin (Biotage, Uppsala, Sweden) were conditioned with 300 mL of methanol and 500 mL of milliQ water.

Article Title: C-Glycosylated flavonoids from black gram husk: Protection against DNA and erythrocytes from oxidative damage and their cytotoxic effect on HeLa cells
Article Snippet: .. 2.5 Purification of vitexin and isovitexin of aqueous ethanol fraction by preparative HPLC The chromatographic separation was performed according to the method described in Section , on a Shimadzu Prep LC8A Preparative Chromatography system equipped with SCL-10AVP system controller (Shimadzu). .. The preparative HPLC was performed on a Varian, Pursuit XRs 10 C18 preparative column (250 mm × 21.2 mm).

Column Chromatography:

Article Title: Antiobesity Effects of Gentiana lutea Extract on 3T3-L1 Preadipocytes and a High-Fat Diet-Induced Mouse Model
Article Snippet: .. Column chromatography (CC) was performed using Silica gel 60 (70~230 mesh, Merck, Darmstadt, Germany), ODS-A (12 nm S-7 μm, YMC GEL, Tokyo, Japan), and Preparative HPLC was performed on LC-8A (Shimadzu, Kyoto, Japan). .. Mouse 3T3-L1 preadipocytes were incubated in Dulbecco’s modified Eagle’s medium supplemented with streptomycin (100 µg/mL), penicillin (100 U/mL), and 10% fetal bovine serum.

Isolation:

Article Title: Methyl Salicylate Glycosides in Some Italian Varietal Wines
Article Snippet: .. Flash Chromatography with Isolute ENV+ and Preparative HPLC for the Isolation of the Single Glycosides A Shimadzu SCL-10 AVP preparative HPLC system with a Shimadzu SPD-10 AVP UV-VIS detector, 8A pumps, and Software Class VP (Shimadzu Corp., Kyoto, Japan) were used for chromatographic separation. .. For flash chromatography, 30 g of ENV+ resin (Biotage, Uppsala, Sweden) were conditioned with 300 mL of methanol and 500 mL of milliQ water.

Purification:

Article Title: C-Glycosylated flavonoids from black gram husk: Protection against DNA and erythrocytes from oxidative damage and their cytotoxic effect on HeLa cells
Article Snippet: .. 2.5 Purification of vitexin and isovitexin of aqueous ethanol fraction by preparative HPLC The chromatographic separation was performed according to the method described in Section , on a Shimadzu Prep LC8A Preparative Chromatography system equipped with SCL-10AVP system controller (Shimadzu). .. The preparative HPLC was performed on a Varian, Pursuit XRs 10 C18 preparative column (250 mm × 21.2 mm).

Article Title: Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿Cloning of Separate Meilingmycin Biosynthesis Gene Clusters by Use of Acyltransferase-Ketoreductase Didomain PCR Amplification ▿ †
Article Snippet: .. Together with fractions 9 and 10, five different meilingmycins were purified by preparative HPLC for the second time with the same condition to yield meilingmycins D (4.7 mg), E (2.9 mg), A (12.4 mg), B (3.9 mg), and C (2.1 mg). .. Nuclear magnetic resonance (NMR) spectra were recorded with a Bruker AM 500 NMR spectrometer in CDCl3 at 25°C, using the solvent as internal reference downfield of tetramethylsilane (TMS) at 0 ppm.

Software:

Article Title: Methyl Salicylate Glycosides in Some Italian Varietal Wines
Article Snippet: .. Flash Chromatography with Isolute ENV+ and Preparative HPLC for the Isolation of the Single Glycosides A Shimadzu SCL-10 AVP preparative HPLC system with a Shimadzu SPD-10 AVP UV-VIS detector, 8A pumps, and Software Class VP (Shimadzu Corp., Kyoto, Japan) were used for chromatographic separation. .. For flash chromatography, 30 g of ENV+ resin (Biotage, Uppsala, Sweden) were conditioned with 300 mL of methanol and 500 mL of milliQ water.

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    Shimadzu Corporation preparative reverse phase hplc
    Time-course of the incubation of oxidized <t>AMC</t> in human serum. Notes: Upper panels of ( A – F ) show the <t>HPLC</t> analysis assessed at 0, 0.5, 2, 3, 6, and 24 hours of incubation, respectively; lower panels of ( A – F ) are respective mass spectra at the corresponding retention time. Abbreviations: AMC, antimicrobial cyclic peptide; HPLC, high-performance liquid chromatography; TIC, total ion current.
    Preparative Reverse Phase Hplc, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/preparative reverse phase hplc/product/Shimadzu Corporation
    Average 90 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
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    89
    Shimadzu Corporation preparative hplc spd 20 a uv dad
    Chromatogram obtained in <t>HPLC-UV-DAD</t> of Cop with wavelength of 257 nm. C 18 column (4.6 × 250 mm, 5 mM), mobile phase: (A) water and 0.01% trifluoroacetic acid and (B) MeOH, gradient elution: 15–50% of B in 45 min, 50–90% of B by 65 min, and 90–15% of B by 70 minutes followed by 2 minutes of 15% B; flow rate: 1 mL/min.
    Preparative Hplc Spd 20 A Uv Dad, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/preparative hplc spd 20 a uv dad/product/Shimadzu Corporation
    Average 89 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
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    92
    Shimadzu Corporation semi preparative hplc
    3D (A) and 2D profiles (B) of E30 fraction on analytic and prepared <t>HPLC-DAD.</t>
    Semi Preparative Hplc, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 92/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/semi preparative hplc/product/Shimadzu Corporation
    Average 92 stars, based on 105 article reviews
    Price from $9.99 to $1999.99
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    92/100 stars
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    88
    Shimadzu Corporation prominence preparative hplc system
    <t>SBE</t> Fraction 4 (F4) showed potent chondroprotective effects in human OA chondrocytes. ( A ) <t>HPLC</t> chromatogram of ethanolic extract of S. baicalensis. Fractions were collected based on indicated retention time. ( B ) SBE fraction (s) was not toxic to human OA chondrocytes. Human OA chondrocytes were treated with indicated concentration (1–50 μg/ml) of SBE fraction (F1 or F2 or F3 or F4) for 24 h and cells viability were measured by MTT assays. Chondrocytes treated with 0.1% DMSO served as control. Viability was expressed relative to control cells. Data points represent mean ± SD from three subjects. ( C ) SBE fraction 4 (F4) inhibited IL-1β induced ROS production in OA chondrocytes. Human OA chondrocytes were treated with SBE fraction (F1 or F2 or F3 or F4) (50 μg/ml) for 2 h, then stained with DHR-123 (5 μM) for 0.5 h at 37 °C, and stimulated with IL-1β (1 ng/ml) for 5 minutes at 37 °C. Fluorescence emission was measured at 535 nm. Bar graph shows relative fluorescence units indicating ROS levels. ( D–E ) SBE fraction 4 (F4) inhibited IL-1β induced mRNA expression of IL-6 and MMP-13 in OA chondrocytes. Primary human OA chondrocytes were pre-treated with SBE fraction (s) for 2 h followed by treatment with IL-1β (1 ng/ml) for 16 h. At the end of treatment chondrocytes were harvested, total RNA were isolated, reverse transcribed to cDNA and mRNA expression of IL-6 ( D ) and MMP-13 ( E ) were measured by quantitative PCR using the TaqMan assay system. Expression of β-actin was used as endogenous expression control. Bar graph represent mean ± SD from three subjects **p ≤ 0.01, as compared to control, # ≤ 0.01, as compared to IL-1β.
    Prominence Preparative Hplc System, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/prominence preparative hplc system/product/Shimadzu Corporation
    Average 88 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
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    Time-course of the incubation of oxidized AMC in human serum. Notes: Upper panels of ( A – F ) show the HPLC analysis assessed at 0, 0.5, 2, 3, 6, and 24 hours of incubation, respectively; lower panels of ( A – F ) are respective mass spectra at the corresponding retention time. Abbreviations: AMC, antimicrobial cyclic peptide; HPLC, high-performance liquid chromatography; TIC, total ion current.

    Journal: International Journal of Nanomedicine

    Article Title: Design and activity of a cyclic mini-β-defensin analog: a novel antimicrobial tool

    doi: 10.2147/IJN.S89610

    Figure Lengend Snippet: Time-course of the incubation of oxidized AMC in human serum. Notes: Upper panels of ( A – F ) show the HPLC analysis assessed at 0, 0.5, 2, 3, 6, and 24 hours of incubation, respectively; lower panels of ( A – F ) are respective mass spectra at the corresponding retention time. Abbreviations: AMC, antimicrobial cyclic peptide; HPLC, high-performance liquid chromatography; TIC, total ion current.

    Article Snippet: The oxidized AMC analog was purified by preparative reverse-phase HPLC (Shimadzu SPD-10A VP UV-Vis detector) using a gradient of acetonitrile (0.1% TFA) in water from 5% to 70% for 20 minutes and a Jupiter column (10 µm, Proteo 90 Å, 250×21 mm).

    Techniques: Incubation, High Performance Liquid Chromatography

    Chromatogram obtained in HPLC-UV-DAD of Cop with wavelength of 257 nm. C 18 column (4.6 × 250 mm, 5 mM), mobile phase: (A) water and 0.01% trifluoroacetic acid and (B) MeOH, gradient elution: 15–50% of B in 45 min, 50–90% of B by 65 min, and 90–15% of B by 70 minutes followed by 2 minutes of 15% B; flow rate: 1 mL/min.

    Journal: BioMed Research International

    Article Title: Antiedematogenic Evaluation of Copaifera langsdorffii Leaves Hydroethanolic Extract and Its Major Compounds

    doi: 10.1155/2015/913152

    Figure Lengend Snippet: Chromatogram obtained in HPLC-UV-DAD of Cop with wavelength of 257 nm. C 18 column (4.6 × 250 mm, 5 mM), mobile phase: (A) water and 0.01% trifluoroacetic acid and (B) MeOH, gradient elution: 15–50% of B in 45 min, 50–90% of B by 65 min, and 90–15% of B by 70 minutes followed by 2 minutes of 15% B; flow rate: 1 mL/min.

    Article Snippet: Two fractions were purified in a preparative HPLC SPD-20 A UV-DAD with software LC-Solution Single (C18 Prep-ODS; 20 mm × 25 cm; Shimadzu) using an elution program consisting of 35–80% CH3 OH in water (v/v) (20 min).

    Techniques: High Performance Liquid Chromatography, Flow Cytometry

    3D (A) and 2D profiles (B) of E30 fraction on analytic and prepared HPLC-DAD.

    Journal: PLoS ONE

    Article Title: An Efficient Preparation of Mulberroside A from the Branch Bark of Mulberry and Its Effect on the Inhibition of Tyrosinase Activity

    doi: 10.1371/journal.pone.0109396

    Figure Lengend Snippet: 3D (A) and 2D profiles (B) of E30 fraction on analytic and prepared HPLC-DAD.

    Article Snippet: The sample preparation was performed using a Semi-Preparative HPLC (Shimadzu LC-6A) equipped with a DAD detector (SPDM20A).

    Techniques: High Performance Liquid Chromatography

    SBE Fraction 4 (F4) showed potent chondroprotective effects in human OA chondrocytes. ( A ) HPLC chromatogram of ethanolic extract of S. baicalensis. Fractions were collected based on indicated retention time. ( B ) SBE fraction (s) was not toxic to human OA chondrocytes. Human OA chondrocytes were treated with indicated concentration (1–50 μg/ml) of SBE fraction (F1 or F2 or F3 or F4) for 24 h and cells viability were measured by MTT assays. Chondrocytes treated with 0.1% DMSO served as control. Viability was expressed relative to control cells. Data points represent mean ± SD from three subjects. ( C ) SBE fraction 4 (F4) inhibited IL-1β induced ROS production in OA chondrocytes. Human OA chondrocytes were treated with SBE fraction (F1 or F2 or F3 or F4) (50 μg/ml) for 2 h, then stained with DHR-123 (5 μM) for 0.5 h at 37 °C, and stimulated with IL-1β (1 ng/ml) for 5 minutes at 37 °C. Fluorescence emission was measured at 535 nm. Bar graph shows relative fluorescence units indicating ROS levels. ( D–E ) SBE fraction 4 (F4) inhibited IL-1β induced mRNA expression of IL-6 and MMP-13 in OA chondrocytes. Primary human OA chondrocytes were pre-treated with SBE fraction (s) for 2 h followed by treatment with IL-1β (1 ng/ml) for 16 h. At the end of treatment chondrocytes were harvested, total RNA were isolated, reverse transcribed to cDNA and mRNA expression of IL-6 ( D ) and MMP-13 ( E ) were measured by quantitative PCR using the TaqMan assay system. Expression of β-actin was used as endogenous expression control. Bar graph represent mean ± SD from three subjects **p ≤ 0.01, as compared to control, # ≤ 0.01, as compared to IL-1β.

    Journal: Scientific Reports

    Article Title: A wogonin-rich-fraction of Scutellaria baicalensis root extract exerts chondroprotective effects by suppressing IL-1β-induced activation of AP-1 in human OA chondrocytes

    doi: 10.1038/srep43789

    Figure Lengend Snippet: SBE Fraction 4 (F4) showed potent chondroprotective effects in human OA chondrocytes. ( A ) HPLC chromatogram of ethanolic extract of S. baicalensis. Fractions were collected based on indicated retention time. ( B ) SBE fraction (s) was not toxic to human OA chondrocytes. Human OA chondrocytes were treated with indicated concentration (1–50 μg/ml) of SBE fraction (F1 or F2 or F3 or F4) for 24 h and cells viability were measured by MTT assays. Chondrocytes treated with 0.1% DMSO served as control. Viability was expressed relative to control cells. Data points represent mean ± SD from three subjects. ( C ) SBE fraction 4 (F4) inhibited IL-1β induced ROS production in OA chondrocytes. Human OA chondrocytes were treated with SBE fraction (F1 or F2 or F3 or F4) (50 μg/ml) for 2 h, then stained with DHR-123 (5 μM) for 0.5 h at 37 °C, and stimulated with IL-1β (1 ng/ml) for 5 minutes at 37 °C. Fluorescence emission was measured at 535 nm. Bar graph shows relative fluorescence units indicating ROS levels. ( D–E ) SBE fraction 4 (F4) inhibited IL-1β induced mRNA expression of IL-6 and MMP-13 in OA chondrocytes. Primary human OA chondrocytes were pre-treated with SBE fraction (s) for 2 h followed by treatment with IL-1β (1 ng/ml) for 16 h. At the end of treatment chondrocytes were harvested, total RNA were isolated, reverse transcribed to cDNA and mRNA expression of IL-6 ( D ) and MMP-13 ( E ) were measured by quantitative PCR using the TaqMan assay system. Expression of β-actin was used as endogenous expression control. Bar graph represent mean ± SD from three subjects **p ≤ 0.01, as compared to control, # ≤ 0.01, as compared to IL-1β.

    Article Snippet: Activity guided fractionation of SBE The extract was fractionated on Prominence preparative HPLC system (Shimadzu Corporation, Kyoto, Japan) and chromatographic separation was performed at 40 °C using a Luna C18 preparative column (5 μm particle size, 150 cm L × 21.20 mm I.D.

    Techniques: High Performance Liquid Chromatography, Concentration Assay, MTT Assay, Staining, Fluorescence, Expressing, Isolation, Real-time Polymerase Chain Reaction, TaqMan Assay