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  • 99
    Thermo Fisher c18 column
    C18 Column, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3761 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c18 column/product/Thermo Fisher
    Average 99 stars, based on 3761 article reviews
    Price from $9.99 to $1999.99
    c18 column - by Bioz Stars, 2020-10
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    92
    Waters Corporation c18 column
    Detection of D3, 7DHC and L3 in honey. Whole honey extract was analyzed directly using a Zorbax Eclipse Plus <t>C18</t> column connected to a Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA) using a methanol gradient. The extracted ion chromatogram (EIC) was obtained using m/z = 407.329 [M + Na] + . Arrow 1, RT of D3 standard; arrow 2, RT of 7DHC standard; arrow 3, RT of L3 standard.
    C18 Column, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 92/100, based on 7364 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c18 column/product/Waters Corporation
    Average 92 stars, based on 7364 article reviews
    Price from $9.99 to $1999.99
    c18 column - by Bioz Stars, 2020-10
    92/100 stars
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    94
    Phenomenex c18 column
    Production of riboflavin in Pseudogymnoascus destructans . ( a ) HPLC chromatogram of SPE <t>C18</t> extract from liquid cultivation medium of P. destructans strain 20631-21 T (see Fig. 6a,b for medium and blank controls). ( b ) Average production curve (±standard error) of riboflavin in six strains of P. destructans ( Fig. 3 ) shows continuous metabolite accumulation. ( c ) Average production curve of [Fe 3+ ] triacetylfusarinine C in six strains of P. destructans peaks after 6 to 8 weeks of culture. The concentration of the metabolite was corrected for biomass content in the fermentation medium.
    C18 Column, supplied by Phenomenex, used in various techniques. Bioz Stars score: 94/100, based on 6929 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c18 column/product/Phenomenex
    Average 94 stars, based on 6929 article reviews
    Price from $9.99 to $1999.99
    c18 column - by Bioz Stars, 2020-10
    94/100 stars
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    99
    Agilent technologies zorbax sb c18 column
    Representative LC-ESI MS/MS MRM (multi reaction monitoring) chromatogram of urophysial sample of round goby. Chromatographic conditions: Agilent <t>Zorbax</t> Extend Plus <t>C18</t> column (50 mm × 2.1 mm I.D., 1.8 μm particle); elution: solvent A (0.1 % acetic acid in H 2 O), solvent B [0.1 % acetic acid in acetonitrile: H 2 O (3:1)], a gradient elution was used starting from 5 to 30 % B in 5.3 min; flow rate 0.6 mL/min; column temperature 20 °C; injection volume 5 μL; the monitored mass transitions for AVT were set at m/z 525.5 → 517.2 and for IT were set at m/z 483.7 → 136.1
    Zorbax Sb C18 Column, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 99/100, based on 5424 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/zorbax sb c18 column/product/Agilent technologies
    Average 99 stars, based on 5424 article reviews
    Price from $9.99 to $1999.99
    zorbax sb c18 column - by Bioz Stars, 2020-10
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    99
    Thermo Fisher hypersil gold c18 column
    Representative LC-ESI MS/MS MRM (multi reaction monitoring) chromatogram of urophysial sample of round goby. Chromatographic conditions: Agilent <t>Zorbax</t> Extend Plus <t>C18</t> column (50 mm × 2.1 mm I.D., 1.8 μm particle); elution: solvent A (0.1 % acetic acid in H 2 O), solvent B [0.1 % acetic acid in acetonitrile: H 2 O (3:1)], a gradient elution was used starting from 5 to 30 % B in 5.3 min; flow rate 0.6 mL/min; column temperature 20 °C; injection volume 5 μL; the monitored mass transitions for AVT were set at m/z 525.5 → 517.2 and for IT were set at m/z 483.7 → 136.1
    Hypersil Gold C18 Column, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 950 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hypersil gold c18 column/product/Thermo Fisher
    Average 99 stars, based on 950 article reviews
    Price from $9.99 to $1999.99
    hypersil gold c18 column - by Bioz Stars, 2020-10
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    92
    Waters Corporation sep pak c18 column
    The R2 fraction prepared from the ethyl acetate fraction of the roasted coffee bean extract exhibits anti-inflammatory activity. ( A ) Procedure for the fractionation of the ethyl acetate fraction of coffee bean extract using the Sep <t>Pak</t> <t>C18</t> column. ( B ) The ethyl acetate fraction of the roasted coffee bean extract ( E ) and the R1-R3 fractions were analyzed by HPLC. ( C – F ) RAW264.7 cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the LPS stimulation (1 μg/mL). ( C ) Nitrate concentrations in culture supernatants were measured 24 h after the LPS stimulation using Griess reagent. ( D ) iNOS mRNA expression was assessed 12 h after the LPS stimulation by RT-PCR. GAPDH mRNA expression was used as an internal control. ( E ) The amounts of CCL2 in supernatants were evaluated 24 h after the LPS stimulation by ELISA. ( F ) CCL2 mRNA expression was assessed 2 h after the LPS stimulation by RT-PCR. ( G ) RAW264.7 cells were transfected with pNF-κB-Luc and pRL-TK. Transfected cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the stimulation with LPS (1 μg/mL) for 6 h. NF-κB-dependent luciferase activity was normalized to the activity of constitutively expressed Renilla luciferase. ( H ) RAW264.7 cells were treated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h. Nuclear extracts were immunoblotted with an anti-Nrf2 or anti-Lamin B antibody. The relative expression levels of Nrf2 in the nucleus are shown in the graph.
    Sep Pak C18 Column, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 92/100, based on 837 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sep pak c18 column/product/Waters Corporation
    Average 92 stars, based on 837 article reviews
    Price from $9.99 to $1999.99
    sep pak c18 column - by Bioz Stars, 2020-10
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    Image Search Results


    Detection of D3, 7DHC and L3 in honey. Whole honey extract was analyzed directly using a Zorbax Eclipse Plus C18 column connected to a Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA) using a methanol gradient. The extracted ion chromatogram (EIC) was obtained using m/z = 407.329 [M + Na] + . Arrow 1, RT of D3 standard; arrow 2, RT of 7DHC standard; arrow 3, RT of L3 standard.

    Journal: Molecules

    Article Title: Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

    doi: 10.3390/molecules25112583

    Figure Lengend Snippet: Detection of D3, 7DHC and L3 in honey. Whole honey extract was analyzed directly using a Zorbax Eclipse Plus C18 column connected to a Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA) using a methanol gradient. The extracted ion chromatogram (EIC) was obtained using m/z = 407.329 [M + Na] + . Arrow 1, RT of D3 standard; arrow 2, RT of 7DHC standard; arrow 3, RT of L3 standard.

    Article Snippet: Liquid Chromatography-Mass Spectrometry (LC-MS) Extracted samples from the local source were redissolved in 10 mL methanol and aliquots analyzed with a qTof LC-MS using Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA, USA), or analyzed after pre-purification carried out using a 1260 Infinity II HPLC system with a C18 column (250 × 4.6 mm, 5 μm particle size) (Waters, Milford, MA, USA).

    Techniques:

    Detection of 1,25(OH) 2 D3 in honey. ( A ) Whole extract was analyzed directly using LC-MS with a Zorbax Eclipse Plus C18 column with a methanol gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + . ( B ) The sample was pre-purified using a C18 column, then analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + .

    Journal: Molecules

    Article Title: Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

    doi: 10.3390/molecules25112583

    Figure Lengend Snippet: Detection of 1,25(OH) 2 D3 in honey. ( A ) Whole extract was analyzed directly using LC-MS with a Zorbax Eclipse Plus C18 column with a methanol gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + . ( B ) The sample was pre-purified using a C18 column, then analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + .

    Article Snippet: Liquid Chromatography-Mass Spectrometry (LC-MS) Extracted samples from the local source were redissolved in 10 mL methanol and aliquots analyzed with a qTof LC-MS using Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA, USA), or analyzed after pre-purification carried out using a 1260 Infinity II HPLC system with a C18 column (250 × 4.6 mm, 5 μm particle size) (Waters, Milford, MA, USA).

    Techniques: Liquid Chromatography with Mass Spectroscopy, Purification

    Detection of 25(OH)D3 in honey. After a pre-purification step using a C18 column with an acetonitrile gradient, the fraction was analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 383.331 [M + H − H 2 O] + .

    Journal: Molecules

    Article Title: Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

    doi: 10.3390/molecules25112583

    Figure Lengend Snippet: Detection of 25(OH)D3 in honey. After a pre-purification step using a C18 column with an acetonitrile gradient, the fraction was analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 383.331 [M + H − H 2 O] + .

    Article Snippet: Liquid Chromatography-Mass Spectrometry (LC-MS) Extracted samples from the local source were redissolved in 10 mL methanol and aliquots analyzed with a qTof LC-MS using Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA, USA), or analyzed after pre-purification carried out using a 1260 Infinity II HPLC system with a C18 column (250 × 4.6 mm, 5 μm particle size) (Waters, Milford, MA, USA).

    Techniques: Purification, Liquid Chromatography with Mass Spectroscopy

    Detection of 1,20(OH) 2 D3 in honey. ( A ) The peak corresponding in RT to standard was collected from commercially available honey using an Atlantis C18 column using a methanol gradient, then analyzed using LC-MS with an ACQUITY UPLC BEH C18 column with a methanol gradient. The EIC was obtained using m/z = 439.319 [M + Na] + . ( B ) The local honey sample was pre-purified using a C18 column, then analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + .

    Journal: Molecules

    Article Title: Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

    doi: 10.3390/molecules25112583

    Figure Lengend Snippet: Detection of 1,20(OH) 2 D3 in honey. ( A ) The peak corresponding in RT to standard was collected from commercially available honey using an Atlantis C18 column using a methanol gradient, then analyzed using LC-MS with an ACQUITY UPLC BEH C18 column with a methanol gradient. The EIC was obtained using m/z = 439.319 [M + Na] + . ( B ) The local honey sample was pre-purified using a C18 column, then analyzed using LC-MS with a Zorbax Eclipse Plus C18 column with an acetonitrile gradient. The EIC was obtained using m/z = 399.326 [M + H − H 2 O] + .

    Article Snippet: Liquid Chromatography-Mass Spectrometry (LC-MS) Extracted samples from the local source were redissolved in 10 mL methanol and aliquots analyzed with a qTof LC-MS using Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA, USA), or analyzed after pre-purification carried out using a 1260 Infinity II HPLC system with a C18 column (250 × 4.6 mm, 5 μm particle size) (Waters, Milford, MA, USA).

    Techniques: Liquid Chromatography with Mass Spectroscopy, Purification

    Detection of 20(OH)D3 and 20(OH)-7DHC in honey. ( A ) After a pre-purification step using an Atlantis C18 column with a methanol gradient, then analyzed using LC-MS with an ACQUITY UPLC BEH C18 column with a methanol gradient. The EIC was obtained using m/z = 423.324 [M + Na] + . ( B ) After extraction of commercially sourced honey, the sample was directly analyzed using LC-MS with a Pursuit 200Å PFP column with a methanol gradient. The EIC was obtained using m/z = 383.331 [M + H − H 2 O] + .

    Journal: Molecules

    Article Title: Detection of 7-Dehydrocholesterol and Vitamin D3 Derivatives in Honey

    doi: 10.3390/molecules25112583

    Figure Lengend Snippet: Detection of 20(OH)D3 and 20(OH)-7DHC in honey. ( A ) After a pre-purification step using an Atlantis C18 column with a methanol gradient, then analyzed using LC-MS with an ACQUITY UPLC BEH C18 column with a methanol gradient. The EIC was obtained using m/z = 423.324 [M + Na] + . ( B ) After extraction of commercially sourced honey, the sample was directly analyzed using LC-MS with a Pursuit 200Å PFP column with a methanol gradient. The EIC was obtained using m/z = 383.331 [M + H − H 2 O] + .

    Article Snippet: Liquid Chromatography-Mass Spectrometry (LC-MS) Extracted samples from the local source were redissolved in 10 mL methanol and aliquots analyzed with a qTof LC-MS using Xevo G2 XS equipped with an ACQUITY UPLC I-Class System (Waters, Milford, MA, USA), or analyzed after pre-purification carried out using a 1260 Infinity II HPLC system with a C18 column (250 × 4.6 mm, 5 μm particle size) (Waters, Milford, MA, USA).

    Techniques: Purification, Liquid Chromatography with Mass Spectroscopy

    Plasmalogen cleavage. Liver homogenates (∼2 mg tissue) (A), brain homogenates (∼2 mg tissue) (B), and 1-O-1’-(Z)-octadecenyl-2-oleoyl- sn -glycero-3-phosphocholine (2 nmol) (C) were pretreated with either HCl or NaCl for 15 min at 65°C before aldehyde derivatization and separation of the fluorescent derivatives by RP-HPLC. Also a blank, containing no tissue or lipid, was analyzed. C16-al and C18-al eluted at 7.4 and 8.8 min, respectively. The drop of the blue line at 7.4 min and 8.8 min in (B) is due to a transient closure of the emission filter to protect the detector against too high intensity. The peak at 7.4 min in (C) is likely due to the presence of a contaminant in the stock solution of 1-O-1’-(Z)-octadecenyl-2-oleoyl- sn -glycero-3-phosphocholine.

    Journal: Journal of Lipid Research

    Article Title: RP-HPLC-fluorescence analysis of aliphatic aldehydes: application to aldehyde-generating enzymes HACL1 and SGPL1

    doi: 10.1194/jlr.D044230

    Figure Lengend Snippet: Plasmalogen cleavage. Liver homogenates (∼2 mg tissue) (A), brain homogenates (∼2 mg tissue) (B), and 1-O-1’-(Z)-octadecenyl-2-oleoyl- sn -glycero-3-phosphocholine (2 nmol) (C) were pretreated with either HCl or NaCl for 15 min at 65°C before aldehyde derivatization and separation of the fluorescent derivatives by RP-HPLC. Also a blank, containing no tissue or lipid, was analyzed. C16-al and C18-al eluted at 7.4 and 8.8 min, respectively. The drop of the blue line at 7.4 min and 8.8 min in (B) is due to a transient closure of the emission filter to protect the detector against too high intensity. The peak at 7.4 min in (C) is likely due to the presence of a contaminant in the stock solution of 1-O-1’-(Z)-octadecenyl-2-oleoyl- sn -glycero-3-phosphocholine.

    Article Snippet: After a 1000 g spin, the supernatant was transferred to inserts of HPLC-vials (Alltech) and 1/50 up to 1/5 of the total volume was injected on a Symmetry® C18 column (4.6 × 150 mm; 5 μm; 100 Å Waters) equilibrated with 80% (v/v) methanol.

    Techniques: High Performance Liquid Chromatography

    Recovery of long-chain aldehydes in presence of proteins. A: Effect of methanol on standards. C13-al (triangles) and C17-al (open squares) (1.5 and 2 nmol, respectively) were derivatized following the optimized protocol in the presence of 33% (v/v) methanol. Upon derivatization, equal volumes of water, water/methanol, or pure methanol were added to the assay to obtain the indicated methanol concentration. After centrifugation, the supernatant was subjected to RP-HPLC, and the fluorescent derivatives were detected by fluorimetry (Ex 390 nm, Em 460 nm). Values represent means of two determinations. B: Effect of methanol on standards in presence of proteins. Brain homogenates (2 mg tissue) were spiked with C13-al (triangles) and C17-al (open squares) before derivatization. Upon derivatization, mixtures were processed as described for (A). Integrated peak areas of C13-al and C17-al derivatives are plotted versus final methanol concentration. C. Recovery of IS in presence of proteins. Increasing amounts of brain and liver homogenates were spiked with C13-al and C17-al before derivatization. Water/methanol or methanol was added to the derivatized sample in order to have a final concentration of either 30% or 60% (v/v). Derivatives were separated by RP-HPLC. Recoveries of C13-al and C17-al derivatives were plotted versus amount of tissue per assay. Open triangles: C13-al, 60% methanol; filled triangles: C13-al, 30% methanol; open squares: C17-al, 60% methanol; filled squares: C17-al, 30% methanol. D. Recovery of IS in presence of proteins. Increasing amounts of brain and liver homogenates were spiked with C13-al (triangles) and C17-al (open squares) before derivatization. Methanol was added to the derivatized sample to a final concentration of 60% (v/v). Derivatives were separated by RP-HPLC. Integrated peak areas of C13-al and C17-al derivatives were plotted versus amount of tissue per assay. E. Analysis of endogenous aldehydes. Increasing amounts of brain and liver homogenates were spiked with C17-al before derivatization and separation of the fluorescent derivatives by RP-HPLC. The ratio of endogenous C16-al (squares) and C18-al (open triangles) over C17-al standard was plotted versus amount of tissue per assay.

    Journal: Journal of Lipid Research

    Article Title: RP-HPLC-fluorescence analysis of aliphatic aldehydes: application to aldehyde-generating enzymes HACL1 and SGPL1

    doi: 10.1194/jlr.D044230

    Figure Lengend Snippet: Recovery of long-chain aldehydes in presence of proteins. A: Effect of methanol on standards. C13-al (triangles) and C17-al (open squares) (1.5 and 2 nmol, respectively) were derivatized following the optimized protocol in the presence of 33% (v/v) methanol. Upon derivatization, equal volumes of water, water/methanol, or pure methanol were added to the assay to obtain the indicated methanol concentration. After centrifugation, the supernatant was subjected to RP-HPLC, and the fluorescent derivatives were detected by fluorimetry (Ex 390 nm, Em 460 nm). Values represent means of two determinations. B: Effect of methanol on standards in presence of proteins. Brain homogenates (2 mg tissue) were spiked with C13-al (triangles) and C17-al (open squares) before derivatization. Upon derivatization, mixtures were processed as described for (A). Integrated peak areas of C13-al and C17-al derivatives are plotted versus final methanol concentration. C. Recovery of IS in presence of proteins. Increasing amounts of brain and liver homogenates were spiked with C13-al and C17-al before derivatization. Water/methanol or methanol was added to the derivatized sample in order to have a final concentration of either 30% or 60% (v/v). Derivatives were separated by RP-HPLC. Recoveries of C13-al and C17-al derivatives were plotted versus amount of tissue per assay. Open triangles: C13-al, 60% methanol; filled triangles: C13-al, 30% methanol; open squares: C17-al, 60% methanol; filled squares: C17-al, 30% methanol. D. Recovery of IS in presence of proteins. Increasing amounts of brain and liver homogenates were spiked with C13-al (triangles) and C17-al (open squares) before derivatization. Methanol was added to the derivatized sample to a final concentration of 60% (v/v). Derivatives were separated by RP-HPLC. Integrated peak areas of C13-al and C17-al derivatives were plotted versus amount of tissue per assay. E. Analysis of endogenous aldehydes. Increasing amounts of brain and liver homogenates were spiked with C17-al before derivatization and separation of the fluorescent derivatives by RP-HPLC. The ratio of endogenous C16-al (squares) and C18-al (open triangles) over C17-al standard was plotted versus amount of tissue per assay.

    Article Snippet: After a 1000 g spin, the supernatant was transferred to inserts of HPLC-vials (Alltech) and 1/50 up to 1/5 of the total volume was injected on a Symmetry® C18 column (4.6 × 150 mm; 5 μm; 100 Å Waters) equilibrated with 80% (v/v) methanol.

    Techniques: Concentration Assay, Centrifugation, High Performance Liquid Chromatography

    Analysis of enzymes producing fatty aldehydes. A: Human recombinant HACL1 (5 μg purified protein) was incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA as substrate (40 μM). A blank reaction was also included, where no substrate was added. The reaction mixtures were fortified with IS (C13-al; 1.5 nmol) before derivatization. B: Human recombinant SGPL1 (0.8 mg protein of soluble E. coli extract) was incubated at 37°C for 60 min with sphinganine-1-phosphate as substrate (40 μM). A blank reaction was also included, where no substrate was added. The reaction mixtures were fortified with IS (C17-al; 2 nmol) before derivatization. C: Liver homogenates of Swiss mice (1 mg tissue, corresponding to ∼0.2 mg protein) were incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA (40 μM), mixtures were spiked with IS (C13-al; 0.74 nmol), and formed aldehydes were analyzed as described in (A). D: Liver homogenates of Swiss mice (2 mg tissue, corresponding to ∼0.4 mg protein) were incubated at 37°C for 60 min with sphinganine-1-phosphate (40 μM), and formed aldehydes were analyzed as described in (B). E: Human fibroblasts were grown either in standard medium (Ct) or in the presence of 1 mM oxythiamine (OT) for two days before HACL1 activity measurement. Fibroblast lysates (0.2–0.5 mg protein) were then incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA (40 μM). Blank reactions were also included, where no substrate was added. The reaction mixtures were spiked with IS (C13-al; 0.74 nmol) before derivatization. Note the presence in fibroblasts of endogenous C16-al and C18-al, eluting at 7.4 and 8.7 min, respectively. F: Lysates of mouse fibroblasts (∼0.2 mg protein), obtained from wild-type or Sgpl1 −/− C57BL6 embryos, were incubated at 37°C for 60 min with sphinganine-1-phosphate (40 μM). Blank reactions were also included, where no substrate was added. The reaction mixtures were spiked with IS (C17-al; 2 nmol) before derivatization, followed by analysis of the generated C16-al. Assays without substrate serve to correct for nonSGPL1-derived C16-al.

    Journal: Journal of Lipid Research

    Article Title: RP-HPLC-fluorescence analysis of aliphatic aldehydes: application to aldehyde-generating enzymes HACL1 and SGPL1

    doi: 10.1194/jlr.D044230

    Figure Lengend Snippet: Analysis of enzymes producing fatty aldehydes. A: Human recombinant HACL1 (5 μg purified protein) was incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA as substrate (40 μM). A blank reaction was also included, where no substrate was added. The reaction mixtures were fortified with IS (C13-al; 1.5 nmol) before derivatization. B: Human recombinant SGPL1 (0.8 mg protein of soluble E. coli extract) was incubated at 37°C for 60 min with sphinganine-1-phosphate as substrate (40 μM). A blank reaction was also included, where no substrate was added. The reaction mixtures were fortified with IS (C17-al; 2 nmol) before derivatization. C: Liver homogenates of Swiss mice (1 mg tissue, corresponding to ∼0.2 mg protein) were incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA (40 μM), mixtures were spiked with IS (C13-al; 0.74 nmol), and formed aldehydes were analyzed as described in (A). D: Liver homogenates of Swiss mice (2 mg tissue, corresponding to ∼0.4 mg protein) were incubated at 37°C for 60 min with sphinganine-1-phosphate (40 μM), and formed aldehydes were analyzed as described in (B). E: Human fibroblasts were grown either in standard medium (Ct) or in the presence of 1 mM oxythiamine (OT) for two days before HACL1 activity measurement. Fibroblast lysates (0.2–0.5 mg protein) were then incubated at 37°C for 5 min with 2-hydroxyoctadecanoyl-CoA (40 μM). Blank reactions were also included, where no substrate was added. The reaction mixtures were spiked with IS (C13-al; 0.74 nmol) before derivatization. Note the presence in fibroblasts of endogenous C16-al and C18-al, eluting at 7.4 and 8.7 min, respectively. F: Lysates of mouse fibroblasts (∼0.2 mg protein), obtained from wild-type or Sgpl1 −/− C57BL6 embryos, were incubated at 37°C for 60 min with sphinganine-1-phosphate (40 μM). Blank reactions were also included, where no substrate was added. The reaction mixtures were spiked with IS (C17-al; 2 nmol) before derivatization, followed by analysis of the generated C16-al. Assays without substrate serve to correct for nonSGPL1-derived C16-al.

    Article Snippet: After a 1000 g spin, the supernatant was transferred to inserts of HPLC-vials (Alltech) and 1/50 up to 1/5 of the total volume was injected on a Symmetry® C18 column (4.6 × 150 mm; 5 μm; 100 Å Waters) equilibrated with 80% (v/v) methanol.

    Techniques: Recombinant, Purification, Incubation, Mouse Assay, Activity Assay, Generated, Derivative Assay

    Ion chromatograms of conjugated BAs on an ACQUITY BEH C18 column eluted by the Gradient-I with various buffers (from bottom to top: 0.01% formic acid, 2 mM ammonium formate at pH 3.5, 2 mM ammonium acetate at pH 4.5, 2 mM ammonium acetate at pH 5.5 and 2 mM ammonium bicarbonate at pH 6.5) and acetonitrile as the aqueous phase and the organic phase respectively.

    Journal: Analytical and bioanalytical chemistry

    Article Title: Factors affecting separation and detection of bile acids by liquid chromatography coupled with mass spectrometry at negative mode

    doi: 10.1007/s00216-017-0489-1

    Figure Lengend Snippet: Ion chromatograms of conjugated BAs on an ACQUITY BEH C18 column eluted by the Gradient-I with various buffers (from bottom to top: 0.01% formic acid, 2 mM ammonium formate at pH 3.5, 2 mM ammonium acetate at pH 4.5, 2 mM ammonium acetate at pH 5.5 and 2 mM ammonium bicarbonate at pH 6.5) and acetonitrile as the aqueous phase and the organic phase respectively.

    Article Snippet: The Gradient-I with a run time of 20 min utilized ACQUITY BEH C18 column (1.7 μm, 100 mm × 2.1 mm) (Waters, Milford, MA, USA) maintained at 45 °C.

    Techniques:

    Ion chromatograms of unconjugated BAs on an ACQUITY BEH C18 column eluted by the Gradient-II with various buffers (from bottom to top: 0.01% formic acid, 2 mM ammonium formate at pH 3.5, 2 mM ammonium acetate at pH 4.5, 2 mM ammonium acetate at pH 5.5, 2 mM ammonium bicarbonate at pH 6.5 and 2 mM ammonium bicarbonate at pH 7.5) and acetonitrile as the aqueous phase and the organic phase respectively.

    Journal: Analytical and bioanalytical chemistry

    Article Title: Factors affecting separation and detection of bile acids by liquid chromatography coupled with mass spectrometry at negative mode

    doi: 10.1007/s00216-017-0489-1

    Figure Lengend Snippet: Ion chromatograms of unconjugated BAs on an ACQUITY BEH C18 column eluted by the Gradient-II with various buffers (from bottom to top: 0.01% formic acid, 2 mM ammonium formate at pH 3.5, 2 mM ammonium acetate at pH 4.5, 2 mM ammonium acetate at pH 5.5, 2 mM ammonium bicarbonate at pH 6.5 and 2 mM ammonium bicarbonate at pH 7.5) and acetonitrile as the aqueous phase and the organic phase respectively.

    Article Snippet: The Gradient-I with a run time of 20 min utilized ACQUITY BEH C18 column (1.7 μm, 100 mm × 2.1 mm) (Waters, Milford, MA, USA) maintained at 45 °C.

    Techniques:

    Retention time of unconjugated BAs eluted by the key intermediate gradient programs during optimization of the Gradient-II on an ACQUITY BEH C18 column. Mobile phase A: 0.01% formic acid (v/v) in water; Mobile phase B: acetonitrile; Blue square: monohydroxyl-BAs (isoLCA/ alloLCA/LCA, blue square); Green diamond: dihydroxyl-BAs (muroCA/βUDCA/UDCA/HDCA/ CDCA/DCA/isoDCA); Red circle: trihydroxyl-BAs (βUCA/UCA/ωMCA/βCA/αMCA/βMCA/HCA/ ACA/ CA).

    Journal: Analytical and bioanalytical chemistry

    Article Title: Factors affecting separation and detection of bile acids by liquid chromatography coupled with mass spectrometry at negative mode

    doi: 10.1007/s00216-017-0489-1

    Figure Lengend Snippet: Retention time of unconjugated BAs eluted by the key intermediate gradient programs during optimization of the Gradient-II on an ACQUITY BEH C18 column. Mobile phase A: 0.01% formic acid (v/v) in water; Mobile phase B: acetonitrile; Blue square: monohydroxyl-BAs (isoLCA/ alloLCA/LCA, blue square); Green diamond: dihydroxyl-BAs (muroCA/βUDCA/UDCA/HDCA/ CDCA/DCA/isoDCA); Red circle: trihydroxyl-BAs (βUCA/UCA/ωMCA/βCA/αMCA/βMCA/HCA/ ACA/ CA).

    Article Snippet: The Gradient-I with a run time of 20 min utilized ACQUITY BEH C18 column (1.7 μm, 100 mm × 2.1 mm) (Waters, Milford, MA, USA) maintained at 45 °C.

    Techniques: High Content Screening

    Retention variations with the pH value of mobile phases for the unconjugated BAs (A, C) and the conjugated BAs (B, D) on BEH C18 column (A, B) and HSS T3 column (C, D).

    Journal: Analytical and bioanalytical chemistry

    Article Title: Factors affecting separation and detection of bile acids by liquid chromatography coupled with mass spectrometry at negative mode

    doi: 10.1007/s00216-017-0489-1

    Figure Lengend Snippet: Retention variations with the pH value of mobile phases for the unconjugated BAs (A, C) and the conjugated BAs (B, D) on BEH C18 column (A, B) and HSS T3 column (C, D).

    Article Snippet: The Gradient-I with a run time of 20 min utilized ACQUITY BEH C18 column (1.7 μm, 100 mm × 2.1 mm) (Waters, Milford, MA, USA) maintained at 45 °C.

    Techniques:

    Processing of proHD5 by trypsin at 30 min and 2 h. HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic

    Journal:

    Article Title: The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability *The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞ ♦

    doi: 10.1074/jbc.M801851200

    Figure Lengend Snippet: Processing of proHD5 by trypsin at 30 min and 2 h. HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic

    Article Snippet: Aliquots of 100 μl were withdrawn and quenched with 20 μl of 10% acetic acid at different time intervals (0.5, 1, 2, and 4 h), of which 60 μl was injected for RP-HPLC analysis on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) running a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic acid.

    Techniques: High Performance Liquid Chromatography

    Comparison of proteolytic susceptibility to trypsin between HD5 (8 h) and E14Q-HD5 (5 min). HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45%

    Journal:

    Article Title: The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability *The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞ ♦

    doi: 10.1074/jbc.M801851200

    Figure Lengend Snippet: Comparison of proteolytic susceptibility to trypsin between HD5 (8 h) and E14Q-HD5 (5 min). HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45%

    Article Snippet: Aliquots of 100 μl were withdrawn and quenched with 20 μl of 10% acetic acid at different time intervals (0.5, 1, 2, and 4 h), of which 60 μl was injected for RP-HPLC analysis on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) running a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic acid.

    Techniques: High Performance Liquid Chromatography

    Processing of E57Q-proHD5 by trypsin at 30 min and 2 h. HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic

    Journal:

    Article Title: The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability *The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞The Conserved Salt Bridge in Human ?-Defensin 5 Is Required for Its Precursor Processing and Proteolytic Stability * S⃞ ♦

    doi: 10.1074/jbc.M801851200

    Figure Lengend Snippet: Processing of E57Q-proHD5 by trypsin at 30 min and 2 h. HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic

    Article Snippet: Aliquots of 100 μl were withdrawn and quenched with 20 μl of 10% acetic acid at different time intervals (0.5, 1, 2, and 4 h), of which 60 μl was injected for RP-HPLC analysis on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) running a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic acid.

    Techniques: High Performance Liquid Chromatography

    HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Combretum aff. laxum. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Journal: Scientia Pharmaceutica

    Article Title: Screening of Panamanian Plant Extracts for Pesticidal Properties and HPLC-Based Identification of Active Compounds

    doi: 10.3797/scipharm.1410-10

    Figure Lengend Snippet: HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Combretum aff. laxum. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Article Snippet: A SunFire C18 column (150 x 30 mm i.d., 5 μm; Waters) connected to a pre-column (10 x 30 mm) was used, at a flow rate of 20 mL/min.

    Techniques: High Performance Liquid Chromatography

    HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Erythroxylum macrophyllum. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Journal: Scientia Pharmaceutica

    Article Title: Screening of Panamanian Plant Extracts for Pesticidal Properties and HPLC-Based Identification of Active Compounds

    doi: 10.3797/scipharm.1410-10

    Figure Lengend Snippet: HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Erythroxylum macrophyllum. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Article Snippet: A SunFire C18 column (150 x 30 mm i.d., 5 μm; Waters) connected to a pre-column (10 x 30 mm) was used, at a flow rate of 20 mL/min.

    Techniques: High Performance Liquid Chromatography

    HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Myrcia splendens. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Journal: Scientia Pharmaceutica

    Article Title: Screening of Panamanian Plant Extracts for Pesticidal Properties and HPLC-Based Identification of Active Compounds

    doi: 10.3797/scipharm.1410-10

    Figure Lengend Snippet: HPLC-DAD chromatograms of the crude extract and its polyamide fractions (PA1–PA5) of Myrcia splendens. SunFire C18 column (150 x 3 mm i.d., 3.5 μm); 5–100% MeCN/0.1% aqueous formic acid in 30min, 0.4 mL/min; detection: 210–700nm, maxplot.

    Article Snippet: A SunFire C18 column (150 x 30 mm i.d., 5 μm; Waters) connected to a pre-column (10 x 30 mm) was used, at a flow rate of 20 mL/min.

    Techniques: High Performance Liquid Chromatography

    A total ion chromatogram of a standard mixture of UTL-5g (at m/z of 271.17 > 109.96), ISOX (at m/z of 128.05 > 109.96), DCA (at m/z of 161.92 > 125.95) and the internal standard zileuton (at m/z of 237.08 > 161.03) at the concentration of 1 μM. Chromatographic separation was performed on a Nova-Pak C18 column (4 μm, 3.9 mm × 150 mm) at 30°C, running with mobile phase consisting of 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B) at the flow rate of 0.2 mL/min. Mobile phase gradient B%(min) was programmed as 10 (0) → 95(3.5) → 95 (19) → 10(19.5) → 10(24). The retention times were 16.86 ± 0.02 for UTL-5g, 11.85 ± 0.01 for ISOX and 14.60 ± 0.02 for DCA, and 13.53 ± 0.02 for internal standard zileuton.

    Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

    Article Title: A liquid chromatography with tandem mass spectrometry method for simultaneous determination of UTL-5g and its metabolites in human plasma

    doi: 10.1016/j.jchromb.2015.04.015

    Figure Lengend Snippet: A total ion chromatogram of a standard mixture of UTL-5g (at m/z of 271.17 > 109.96), ISOX (at m/z of 128.05 > 109.96), DCA (at m/z of 161.92 > 125.95) and the internal standard zileuton (at m/z of 237.08 > 161.03) at the concentration of 1 μM. Chromatographic separation was performed on a Nova-Pak C18 column (4 μm, 3.9 mm × 150 mm) at 30°C, running with mobile phase consisting of 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B) at the flow rate of 0.2 mL/min. Mobile phase gradient B%(min) was programmed as 10 (0) → 95(3.5) → 95 (19) → 10(19.5) → 10(24). The retention times were 16.86 ± 0.02 for UTL-5g, 11.85 ± 0.01 for ISOX and 14.60 ± 0.02 for DCA, and 13.53 ± 0.02 for internal standard zileuton.

    Article Snippet: UTL-5g, its metabolites DCA and ISOX, and the internal standard zileuton were separated from potentially interfering material on a Nova-Pak C18 column (4 μm, 3.9 mm × 150 mm; Waters Corp., Milford, MA) maintained at 30 °C.

    Techniques: Concentration Assay, Flow Cytometry

    Production of riboflavin in Pseudogymnoascus destructans . ( a ) HPLC chromatogram of SPE C18 extract from liquid cultivation medium of P. destructans strain 20631-21 T (see Fig. 6a,b for medium and blank controls). ( b ) Average production curve (±standard error) of riboflavin in six strains of P. destructans ( Fig. 3 ) shows continuous metabolite accumulation. ( c ) Average production curve of [Fe 3+ ] triacetylfusarinine C in six strains of P. destructans peaks after 6 to 8 weeks of culture. The concentration of the metabolite was corrected for biomass content in the fermentation medium.

    Journal: Scientific Reports

    Article Title: Vitamin B2 as a virulence factor in Pseudogymnoascus destructans skin infection

    doi: 10.1038/srep33200

    Figure Lengend Snippet: Production of riboflavin in Pseudogymnoascus destructans . ( a ) HPLC chromatogram of SPE C18 extract from liquid cultivation medium of P. destructans strain 20631-21 T (see Fig. 6a,b for medium and blank controls). ( b ) Average production curve (±standard error) of riboflavin in six strains of P. destructans ( Fig. 3 ) shows continuous metabolite accumulation. ( c ) Average production curve of [Fe 3+ ] triacetylfusarinine C in six strains of P. destructans peaks after 6 to 8 weeks of culture. The concentration of the metabolite was corrected for biomass content in the fermentation medium.

    Article Snippet: A Gemini 5 μm C18 column (250 × 4.6 mm, Phenomenex, Torrance, CA, USA) with a guard column was used for the analysis.

    Techniques: High Performance Liquid Chromatography, Concentration Assay

    Production and tolerance of riboflavin in Pseudogymnoascus fungi. ( a ) HPLC chromatogram of SPE C18 extract from liquid cultivation medium of Pseudogymnoascus sp. strain CCF5026. ( b ) Blank liquid cultivation medium HPLC chromatogram. ( c ) Average production curves (±standard error) of riboflavin in non-pathogenic Pseudogymnoascus strains. The concentration of the metabolite was corrected for biomass content in the fermentation medium. ( d ) Tolerance of P. destructans strains to riboflavin in concentrations ranging from 0 to 200 μg ml −1 . Riboflavin was added after autoclaving the medium containing glucose (20 g l −1 ), yeast extract (5 g l −1 ), and agar (20 g l −1 ). Colony diameter was measured after 49 days. Three replicates were used in each experiment, and their values may overlap.

    Journal: Scientific Reports

    Article Title: Vitamin B2 as a virulence factor in Pseudogymnoascus destructans skin infection

    doi: 10.1038/srep33200

    Figure Lengend Snippet: Production and tolerance of riboflavin in Pseudogymnoascus fungi. ( a ) HPLC chromatogram of SPE C18 extract from liquid cultivation medium of Pseudogymnoascus sp. strain CCF5026. ( b ) Blank liquid cultivation medium HPLC chromatogram. ( c ) Average production curves (±standard error) of riboflavin in non-pathogenic Pseudogymnoascus strains. The concentration of the metabolite was corrected for biomass content in the fermentation medium. ( d ) Tolerance of P. destructans strains to riboflavin in concentrations ranging from 0 to 200 μg ml −1 . Riboflavin was added after autoclaving the medium containing glucose (20 g l −1 ), yeast extract (5 g l −1 ), and agar (20 g l −1 ). Colony diameter was measured after 49 days. Three replicates were used in each experiment, and their values may overlap.

    Article Snippet: A Gemini 5 μm C18 column (250 × 4.6 mm, Phenomenex, Torrance, CA, USA) with a guard column was used for the analysis.

    Techniques: High Performance Liquid Chromatography, Concentration Assay

    Enzymatic transformation of Lac-Enk to Gal-Lac-Enk. A) RP-HPLC analysis of one-pot reaction for Lac-Enk with UDP-Glc and galactose epimerase. From top to bottom: substrate (Lac-Enk) only; positive control – reaction mixture containing UDP-Gal and LgtC; negative control – reaction mixture containing UDP-Glc and LgtC; reaction mixture containing UDP-Glc, LgtC and galactose epimerase. All reactions were conducted in 50 mM HEPES (pH 7), 10 mM MnCl 2 buffer at 37°C. RP-HPLC was conducted on an Alltima C18 column using a gradient of 0–90% solvent B, t R = 17.2 min (substrate, Lac-Enk) and t R = 16.8 min (product, Gal-Lac-Enk). B) ES-MS analysis of the acceptor, Lac-Enk m/z (C 44 H 63 N 7 O 18 , 977.4) m/z 978.5 [M+H] + (calcd. 978.4). C) ES-MS analysis of the product, Gal-Lac-Enk (C 52 H 76 N 8 O 23 , 1139.5) m/z 1140.5 [M+H] + (calcd. 1140.5).

    Journal: PLoS ONE

    Article Title: A Drug Delivery Strategy: Binding Enkephalin to Asialoglycoprotein Receptor by Enzymatic Galactosylation

    doi: 10.1371/journal.pone.0095024

    Figure Lengend Snippet: Enzymatic transformation of Lac-Enk to Gal-Lac-Enk. A) RP-HPLC analysis of one-pot reaction for Lac-Enk with UDP-Glc and galactose epimerase. From top to bottom: substrate (Lac-Enk) only; positive control – reaction mixture containing UDP-Gal and LgtC; negative control – reaction mixture containing UDP-Glc and LgtC; reaction mixture containing UDP-Glc, LgtC and galactose epimerase. All reactions were conducted in 50 mM HEPES (pH 7), 10 mM MnCl 2 buffer at 37°C. RP-HPLC was conducted on an Alltima C18 column using a gradient of 0–90% solvent B, t R = 17.2 min (substrate, Lac-Enk) and t R = 16.8 min (product, Gal-Lac-Enk). B) ES-MS analysis of the acceptor, Lac-Enk m/z (C 44 H 63 N 7 O 18 , 977.4) m/z 978.5 [M+H] + (calcd. 978.4). C) ES-MS analysis of the product, Gal-Lac-Enk (C 52 H 76 N 8 O 23 , 1139.5) m/z 1140.5 [M+H] + (calcd. 1140.5).

    Article Snippet: A Phenomenex Luna C18 column (5 µm, 50×2.0 mm, Torrance, USA) was used for LC-MS analysis.

    Techniques: Transformation Assay, High Performance Liquid Chromatography, Gas Chromatography, Positive Control, Negative Control, Mass Spectrometry

    Stability profiles of enkephalin and carbohydrate-derived enkephalins. A) Plasma stability assay: the concentration of compounds in plasma was determined at various time points using RP-HPLC on a C18 Vydac column, gradient of 20–35% (enkephalin) or 0–70% solvent B (glycosylated enkephalins) over 30 min. B) Caco-2 cell stability assay: The concentration of compound in Caco-2 cell homogenates at various time points was quantified using LC-MS. Data presented is the mean ± S.D. (n = 3).

    Journal: PLoS ONE

    Article Title: A Drug Delivery Strategy: Binding Enkephalin to Asialoglycoprotein Receptor by Enzymatic Galactosylation

    doi: 10.1371/journal.pone.0095024

    Figure Lengend Snippet: Stability profiles of enkephalin and carbohydrate-derived enkephalins. A) Plasma stability assay: the concentration of compounds in plasma was determined at various time points using RP-HPLC on a C18 Vydac column, gradient of 20–35% (enkephalin) or 0–70% solvent B (glycosylated enkephalins) over 30 min. B) Caco-2 cell stability assay: The concentration of compound in Caco-2 cell homogenates at various time points was quantified using LC-MS. Data presented is the mean ± S.D. (n = 3).

    Article Snippet: A Phenomenex Luna C18 column (5 µm, 50×2.0 mm, Torrance, USA) was used for LC-MS analysis.

    Techniques: Derivative Assay, Stability Assay, Concentration Assay, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy

    Phytochemical characterization of water extracts of D. morbifera (DM) by HPLC analysis. (A) HPLC-UV chromatogram for four standard compounds (syringin, chlorogenic acid, rutin, and quercetin), (B) HPLC-UV chromatogram for water extracts of D. morbifera (DM), (C) Chemical structures of four major compounds in DM, (D) Regression equations and correlation coefficients for four standard compounds. HPLC analysis was carried out using a gradient elution on a Phenomenex Kinetex C18 column (5 µ, 100 A, 150 × 4.6 mm). The flow rate, column oven temperature, and UV wavelength for detection were set at 1 mL/min, 30 °C, and 254 nm, respectively.

    Journal: International Journal of Biological Sciences

    Article Title: Dendropanax morbifera Ameliorates Thioacetamide-Induced Hepatic Fibrosis via TGF-β1/Smads Pathways

    doi: 10.7150/ijbs.30356

    Figure Lengend Snippet: Phytochemical characterization of water extracts of D. morbifera (DM) by HPLC analysis. (A) HPLC-UV chromatogram for four standard compounds (syringin, chlorogenic acid, rutin, and quercetin), (B) HPLC-UV chromatogram for water extracts of D. morbifera (DM), (C) Chemical structures of four major compounds in DM, (D) Regression equations and correlation coefficients for four standard compounds. HPLC analysis was carried out using a gradient elution on a Phenomenex Kinetex C18 column (5 µ, 100 A, 150 × 4.6 mm). The flow rate, column oven temperature, and UV wavelength for detection were set at 1 mL/min, 30 °C, and 254 nm, respectively.

    Article Snippet: HPLC analysis was carried out by using a Knauer Smartline system (a Manager 5000, two Pump 1000 and an UV Detector 2500) equipped with a Phenomenex Kinetex C18 column (5 µm, 100 A, 150 × 4.6 mm).

    Techniques: High Performance Liquid Chromatography, Flow Cytometry

    (a–l) Separation of prednisolone and impurity A and B peaks obtained with the official Ph. Eur. method using different columns: (a) Venusil AQ C18, (b) Gemini C18, (c) Synergi Hydro-RP, (d) Luna C18(2), (e) Gemini NX-C18, (f) Luna Omega Polar C18, (g) Kinetex F5, (h) Kinetex Biphenyl, (i) Kinetex C18, (j) Kinetex Phenyl-Hexyl, (k) Kinetex XB-C18, and (l) Kinetex Polar C18 ( Table 2 ). All of the chromatograms are on the same y -axis scale and have a 5 min time interval.

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: (a–l) Separation of prednisolone and impurity A and B peaks obtained with the official Ph. Eur. method using different columns: (a) Venusil AQ C18, (b) Gemini C18, (c) Synergi Hydro-RP, (d) Luna C18(2), (e) Gemini NX-C18, (f) Luna Omega Polar C18, (g) Kinetex F5, (h) Kinetex Biphenyl, (i) Kinetex C18, (j) Kinetex Phenyl-Hexyl, (k) Kinetex XB-C18, and (l) Kinetex Polar C18 ( Table 2 ). All of the chromatograms are on the same y -axis scale and have a 5 min time interval.

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    Separation of the prednisolone ( t R = 12.972 min), impurity A ( t R = 13.511 min), and impurity B ( t R = 11.963 min) peaks obtained with the adjusted official Ph. Eur. method using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) given in Table 3 . (The y -scale is the same as in Figure 4 .)

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: Separation of the prednisolone ( t R = 12.972 min), impurity A ( t R = 13.511 min), and impurity B ( t R = 11.963 min) peaks obtained with the adjusted official Ph. Eur. method using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) given in Table 3 . (The y -scale is the same as in Figure 4 .)

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    Chromatogram of prednisolone FSS obtained using the Venusil AQ C18 column (150 mm × 4.6 mm, 3 μm) with the official Ph. Eur. method ( Table 1 ).

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: Chromatogram of prednisolone FSS obtained using the Venusil AQ C18 column (150 mm × 4.6 mm, 3 μm) with the official Ph. Eur. method ( Table 1 ).

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    (a, b) Chromatograms of (a) prednisolone FSS and (b) prednisolone FPI obtained using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) with the improved method reported in Table 5 .

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: (a, b) Chromatograms of (a) prednisolone FSS and (b) prednisolone FPI obtained using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) with the improved method reported in Table 5 .

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    Chromatogram of prednisolone FPI obtained using the Venusil AQ C18 column (150 mm × 4.6 mm, 3 μm) with the official Ph. Eur. method ( Table 1 ).

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: Chromatogram of prednisolone FPI obtained using the Venusil AQ C18 column (150 mm × 4.6 mm, 3 μm) with the official Ph. Eur. method ( Table 1 ).

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    (a, b) Chromatograms of (a) prednisolone FSS and (b) prednisolone FPI obtained using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) with the isocratic method in Table 4 .

    Journal: ACS Omega

    Article Title: An Improved Reversed-Phase High-Performance Liquid Chromatography Method for the Analysis of Related Substances of Prednisolone in Active Ingredient

    doi: 10.1021/acsomega.0c00037

    Figure Lengend Snippet: (a, b) Chromatograms of (a) prednisolone FSS and (b) prednisolone FPI obtained using the Gemini C18 column (150 mm × 4.6 mm, 3 μm) with the isocratic method in Table 4 .

    Article Snippet: The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm.

    Techniques:

    Representative radio-HPLC traces for 30 min p.i. of the metabolism study of ( S )-[ 18 F] 9a . The radiochemical purity of ( S )-[ 18 F] 9a was > 97% before injection. The samples were analyzed by HPLC, using a γ-detector (Raytest Isotopenmessgeräte GmbH/Agilent). The HPLC was done on a Phenomenex C18 column (250 × 4.6 mm) using a gradient method with acetonitrile and water (both having 0.05% TFA)

    Journal: EJNMMI Radiopharmacy and Chemistry

    Article Title: Synthesis, radiosynthesis, in vitro and first in vivo evaluation of a new matrix metalloproteinase inhibitor based on γ-fluorinated α-sulfonylaminohydroxamic acid

    doi: 10.1186/s41181-018-0045-0

    Figure Lengend Snippet: Representative radio-HPLC traces for 30 min p.i. of the metabolism study of ( S )-[ 18 F] 9a . The radiochemical purity of ( S )-[ 18 F] 9a was > 97% before injection. The samples were analyzed by HPLC, using a γ-detector (Raytest Isotopenmessgeräte GmbH/Agilent). The HPLC was done on a Phenomenex C18 column (250 × 4.6 mm) using a gradient method with acetonitrile and water (both having 0.05% TFA)

    Article Snippet: HPLC was done on a Phenomenex C18 column (250 mm × 4.6 mm) using a gradient method with acetonitrile and water (both having 0.05% TFA).

    Techniques: High Performance Liquid Chromatography, Injection

    HPLC-FLD chromatograms after periodate oxidation from: a) overlaid toxins standards of dcNEO (0.37 μM) and dcSTX (0.70 μM), both producing two oxidation products coded ‘1’ and ‘2’; b) urine sample from patient A on 2018-10-11 after C18 cleanup dominated by dcSTX oxidation products.

    Journal: Toxicon: X

    Article Title: Paralytic shellfish poisoning due to ingestion of contaminated mussels: A 2018 case report in Caparica (Portugal)

    doi: 10.1016/j.toxcx.2019.100017

    Figure Lengend Snippet: HPLC-FLD chromatograms after periodate oxidation from: a) overlaid toxins standards of dcNEO (0.37 μM) and dcSTX (0.70 μM), both producing two oxidation products coded ‘1’ and ‘2’; b) urine sample from patient A on 2018-10-11 after C18 cleanup dominated by dcSTX oxidation products.

    Article Snippet: Separation was performed on a Supelcosil LC-18, 150 × 4.6 mm, 5 μm column (Supelco, USA) equipped with a C18 guard column (SecurityGuard™, 4 × 3 mm, Luna, Phenomenex, USA).

    Techniques: High Performance Liquid Chromatography

    Representative LC-ESI MS/MS MRM (multi reaction monitoring) chromatogram of urophysial sample of round goby. Chromatographic conditions: Agilent Zorbax Extend Plus C18 column (50 mm × 2.1 mm I.D., 1.8 μm particle); elution: solvent A (0.1 % acetic acid in H 2 O), solvent B [0.1 % acetic acid in acetonitrile: H 2 O (3:1)], a gradient elution was used starting from 5 to 30 % B in 5.3 min; flow rate 0.6 mL/min; column temperature 20 °C; injection volume 5 μL; the monitored mass transitions for AVT were set at m/z 525.5 → 517.2 and for IT were set at m/z 483.7 → 136.1

    Journal: Fish Physiology and Biochemistry

    Article Title: Neuropeptides isotocin and arginine vasotocin in urophysis of three fish species

    doi: 10.1007/s10695-012-9746-6

    Figure Lengend Snippet: Representative LC-ESI MS/MS MRM (multi reaction monitoring) chromatogram of urophysial sample of round goby. Chromatographic conditions: Agilent Zorbax Extend Plus C18 column (50 mm × 2.1 mm I.D., 1.8 μm particle); elution: solvent A (0.1 % acetic acid in H 2 O), solvent B [0.1 % acetic acid in acetonitrile: H 2 O (3:1)], a gradient elution was used starting from 5 to 30 % B in 5.3 min; flow rate 0.6 mL/min; column temperature 20 °C; injection volume 5 μL; the monitored mass transitions for AVT were set at m/z 525.5 → 517.2 and for IT were set at m/z 483.7 → 136.1

    Article Snippet: Chromatographic separation was achieved on Agilent Zorbax Eclipse XDB-C18 column (150 mm × 4.6 mm I.D., 5 μm particle size).

    Techniques: Mass Spectrometry, Flow Cytometry, Injection

    Representative HPLC-FL chromatograms of a urophysial sample of round goby and b the same sample spiked with standard AVT (6.6 pmol/mL) and IT (6.8 pmol/mL). Chromatographic conditions: Agilent Zorbax Eclipse XDB-C18 column (150 mm × 4.6 mm I.D., 5 μm particle); elution: solvent A (0.1 % TFA in H 2 O), solvent B [0.1 % TFA in acetonitrile : H 2 O (3:1)], linear gradient 45–80 % of eluent B in 12 min; flow rate 1 mL/min; column temperature 20 °C; injection volume 40 μL; detection: FL, excitation 470 nm, emission 530 nm

    Journal: Fish Physiology and Biochemistry

    Article Title: Neuropeptides isotocin and arginine vasotocin in urophysis of three fish species

    doi: 10.1007/s10695-012-9746-6

    Figure Lengend Snippet: Representative HPLC-FL chromatograms of a urophysial sample of round goby and b the same sample spiked with standard AVT (6.6 pmol/mL) and IT (6.8 pmol/mL). Chromatographic conditions: Agilent Zorbax Eclipse XDB-C18 column (150 mm × 4.6 mm I.D., 5 μm particle); elution: solvent A (0.1 % TFA in H 2 O), solvent B [0.1 % TFA in acetonitrile : H 2 O (3:1)], linear gradient 45–80 % of eluent B in 12 min; flow rate 1 mL/min; column temperature 20 °C; injection volume 40 μL; detection: FL, excitation 470 nm, emission 530 nm

    Article Snippet: Chromatographic separation was achieved on Agilent Zorbax Eclipse XDB-C18 column (150 mm × 4.6 mm I.D., 5 μm particle size).

    Techniques: High Performance Liquid Chromatography, Flow Cytometry, Injection

    HPLC analysis of a) [ 18 F]NST732 after formulation; b) [ 18 F]NST732 co-injected with the non-radioactive standard. HPLC conditions: Agilent Eclipse XDB C18 4.6 × 150 mm, 5 μm column, 20–40% acetonitrile in water (0.1% TFA) for 10

    Journal: Nuclear Medicine and Biology

    Article Title: Synthesis of ApoSense compound [18F]2-(5-(dimethylamino)naphthalene-1-sulfonamido)-2-(fluoromethyl)butanoic acid ([18F]NST732) by nucleophilic ring-opening of an aziridine precursor

    doi: 10.1016/j.nucmedbio.2011.12.008

    Figure Lengend Snippet: HPLC analysis of a) [ 18 F]NST732 after formulation; b) [ 18 F]NST732 co-injected with the non-radioactive standard. HPLC conditions: Agilent Eclipse XDB C18 4.6 × 150 mm, 5 μm column, 20–40% acetonitrile in water (0.1% TFA) for 10

    Article Snippet: Analytical HPLC analyses for radiochemical work were performed on an Agilent 1200 Series instrument equipped with multi-wavelength detectors using an Agilent Eclipse XDB C18 column (4.6 × 150 mm, 5 μm) with a flow rate of 1.0 mL/min.

    Techniques: High Performance Liquid Chromatography, Injection

    HPLC analysis of the crude reaction mixture of compound 5 (t R = 7.3 min) and its regioisomer ( 6 , t R = 8.7 min). HPLC conditions: Agilent Eclipse XDB C18 5 μm, 4.6 × 150 mm column, 35–50% acetonitrile in water (0.1% TFA) in 10 min,

    Journal: Nuclear Medicine and Biology

    Article Title: Synthesis of ApoSense compound [18F]2-(5-(dimethylamino)naphthalene-1-sulfonamido)-2-(fluoromethyl)butanoic acid ([18F]NST732) by nucleophilic ring-opening of an aziridine precursor

    doi: 10.1016/j.nucmedbio.2011.12.008

    Figure Lengend Snippet: HPLC analysis of the crude reaction mixture of compound 5 (t R = 7.3 min) and its regioisomer ( 6 , t R = 8.7 min). HPLC conditions: Agilent Eclipse XDB C18 5 μm, 4.6 × 150 mm column, 35–50% acetonitrile in water (0.1% TFA) in 10 min,

    Article Snippet: Analytical HPLC analyses for radiochemical work were performed on an Agilent 1200 Series instrument equipped with multi-wavelength detectors using an Agilent Eclipse XDB C18 column (4.6 × 150 mm, 5 μm) with a flow rate of 1.0 mL/min.

    Techniques: High Performance Liquid Chromatography

    HPLC analysis of the crude reaction mixture of [ 18 F]NST732 (t R = 7.4 min) and its regioisomer ( 7 , t R = 6.0 min). HPLC conditions: Agilent Eclipse XDB C18 4.6 × 150 mm, 5 μm column, 20–40% acetonitrile in water (0.1% TFA) for 10

    Journal: Nuclear Medicine and Biology

    Article Title: Synthesis of ApoSense compound [18F]2-(5-(dimethylamino)naphthalene-1-sulfonamido)-2-(fluoromethyl)butanoic acid ([18F]NST732) by nucleophilic ring-opening of an aziridine precursor

    doi: 10.1016/j.nucmedbio.2011.12.008

    Figure Lengend Snippet: HPLC analysis of the crude reaction mixture of [ 18 F]NST732 (t R = 7.4 min) and its regioisomer ( 7 , t R = 6.0 min). HPLC conditions: Agilent Eclipse XDB C18 4.6 × 150 mm, 5 μm column, 20–40% acetonitrile in water (0.1% TFA) for 10

    Article Snippet: Analytical HPLC analyses for radiochemical work were performed on an Agilent 1200 Series instrument equipped with multi-wavelength detectors using an Agilent Eclipse XDB C18 column (4.6 × 150 mm, 5 μm) with a flow rate of 1.0 mL/min.

    Techniques: High Performance Liquid Chromatography

    The R2 fraction prepared from the ethyl acetate fraction of the roasted coffee bean extract exhibits anti-inflammatory activity. ( A ) Procedure for the fractionation of the ethyl acetate fraction of coffee bean extract using the Sep Pak C18 column. ( B ) The ethyl acetate fraction of the roasted coffee bean extract ( E ) and the R1-R3 fractions were analyzed by HPLC. ( C – F ) RAW264.7 cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the LPS stimulation (1 μg/mL). ( C ) Nitrate concentrations in culture supernatants were measured 24 h after the LPS stimulation using Griess reagent. ( D ) iNOS mRNA expression was assessed 12 h after the LPS stimulation by RT-PCR. GAPDH mRNA expression was used as an internal control. ( E ) The amounts of CCL2 in supernatants were evaluated 24 h after the LPS stimulation by ELISA. ( F ) CCL2 mRNA expression was assessed 2 h after the LPS stimulation by RT-PCR. ( G ) RAW264.7 cells were transfected with pNF-κB-Luc and pRL-TK. Transfected cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the stimulation with LPS (1 μg/mL) for 6 h. NF-κB-dependent luciferase activity was normalized to the activity of constitutively expressed Renilla luciferase. ( H ) RAW264.7 cells were treated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h. Nuclear extracts were immunoblotted with an anti-Nrf2 or anti-Lamin B antibody. The relative expression levels of Nrf2 in the nucleus are shown in the graph.

    Journal: Scientific Reports

    Article Title: Pyrocatechol, a component of coffee, suppresses LPS-induced inflammatory responses by inhibiting NF-κB and activating Nrf2

    doi: 10.1038/s41598-020-59380-x

    Figure Lengend Snippet: The R2 fraction prepared from the ethyl acetate fraction of the roasted coffee bean extract exhibits anti-inflammatory activity. ( A ) Procedure for the fractionation of the ethyl acetate fraction of coffee bean extract using the Sep Pak C18 column. ( B ) The ethyl acetate fraction of the roasted coffee bean extract ( E ) and the R1-R3 fractions were analyzed by HPLC. ( C – F ) RAW264.7 cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the LPS stimulation (1 μg/mL). ( C ) Nitrate concentrations in culture supernatants were measured 24 h after the LPS stimulation using Griess reagent. ( D ) iNOS mRNA expression was assessed 12 h after the LPS stimulation by RT-PCR. GAPDH mRNA expression was used as an internal control. ( E ) The amounts of CCL2 in supernatants were evaluated 24 h after the LPS stimulation by ELISA. ( F ) CCL2 mRNA expression was assessed 2 h after the LPS stimulation by RT-PCR. ( G ) RAW264.7 cells were transfected with pNF-κB-Luc and pRL-TK. Transfected cells were pretreated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h prior to the stimulation with LPS (1 μg/mL) for 6 h. NF-κB-dependent luciferase activity was normalized to the activity of constitutively expressed Renilla luciferase. ( H ) RAW264.7 cells were treated with fractions R1-R3, equivalent to 5% (v/v) of coffee, for 1 h. Nuclear extracts were immunoblotted with an anti-Nrf2 or anti-Lamin B antibody. The relative expression levels of Nrf2 in the nucleus are shown in the graph.

    Article Snippet: To separate ethyl acetate extracts, dried ethyl acetate extracts (equivalent to 15 mL coffee) were dissolved in 15 mL of methanol/ethyl acetate (2:1), applied to a Sep-Pak C18 column (Waters, Milford, MA), and eluted by 15-mL stepwise rinses of water (R1), 20% methanol (R2), and 100% methanol (R3).

    Techniques: Activity Assay, Fractionation, High Performance Liquid Chromatography, Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Transfection, Luciferase