Bds hypersil c18 hplc column

Manufactured by Thermo Fisher Scientific

Sourced in United States

The BDS Hypersil™ C18 HPLC column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. It features a bonded C18 stationary phase on a high-purity silica support.

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Lab products found in correlation

1525 binary hplc pump, by Waters Corporation (4 mentions) 717 plus autosampler, by Waters Corporation (4 mentions) Hplc system, by Waters Corporation (3 mentions) Hplc ms, by Shimadzu (1 mentions) Ltq ft ultra, by Thermo Fisher Scientific (1 mentions) Du 65 spectrophotometer, by Beckman Coulter (1 mentions) G1367b autosampler, by Agilent Technologies (1 mentions) Waters hplc system, by Waters Corporation (1 mentions)

Close spelling variants of this product

C18 column (178 citations) Hypersil BDS C18 column (74 citations) Hypersil BDS C18 (49 citations) Hypersil C18 column (19 citations) BDS HYPERSIL™ (9 citations) BDS C18 column (5 citations) C18 HPLC column (3 citations)

10 protocols using bds hypersil c18 hplc column

HPLC analysis of potassium phosphate buffer

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Potassium phosphate monobasic buffer (0.2 M) was prepared by dissolving 27.2 g of KH₂PO₄ in 800 mL water, adjusting the pH at 2.8 ± 0.05 with 85% H₃PO₄ and adding water to the total volume of 1000 mL. SOS solution (20 mM) was prepared by dissolving 4.7 g SOS in 1000 mL water. Mobile phases A and B, both consisting of ACN, 20 mM SOS, 0.2 M KH₂PO₄ buffer at pH 2.8 and water were prepared as specified in the Ph. Int. in the following ratios—mobile phase A: ACN/SOS/KH₂PO₄ buffer/water = 4/70/10/16 v/v/v/v; and mobile phase B: ACN/SOS/KH₂PO₄ buffer/water = 17/70/10/3 v/v/v/v.
A Hypersil BDS C18 HPLC column (250 × 4.6 mm; 5 µm) from Thermo Fischer Scientific (San Jose, CA, USA) was used for the analysis. Gradient elution was applied at 45 °C and a flow rate of 1 mL min⁻¹: 0% B (0–16 min), 0%–100%B (16–18 min), 100% B (18–22 min), 100%–0%B (22–24 min), and 0% B (24–30 min). The detection wavelength was set at 219 nm and the injection volume was 50 µL.

Makuc D., Švab Ž., Naumoska K., Plavec J, & Časar Z. (2020). Determination of d-Cycloserine Impurities in Pharmaceutical Dosage Forms: Comparison of the International Pharmacopoeia HPLC–UV Method and the DOSY NMR Method. Molecules, 25(7), 1684.

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Quantitative Analysis of 2,3-BDO and Intracellular Metabolites

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The extracellular 2,3-BDO titer was analyzed by GC-BID (Shimatsu, Japan) using the TG-WAXMS column (Shimatsu). The cells were cultured in 40 ml medium in the shaker flask for 11 days. After harvesting and centrifugation (6,000×g, 15 min), the supernatant was diluted 10-fold and analyzed using GC-BID. Meanwhile, 2,3-BDO standard (Sigma) was diluted to 200, 100, 50, 25, and 10 mg/L and analyzed similarly to establish the standard curve. The 2,3-BDO titers were determined based on the area of the signal and the standard curve.
For the analysis of intracellular metabolites F6P, AcCoA, OAA, and sucrose, the cells were cultured in 40 ml BG-11 medium for 11 days and 1 ml of the supernatant was withdrawn. After centrifugation, the cells were lysed using 100 μL TE buffer containing 2 mg/ml lysozyme in a 37 °C water bath for 20 min, followed by centrifugation (12,000×g, 5 min). Subsequently, 500 μL supernatant was analyzed by HPLC-MS (Shimadzu) using Hypersil™ BDS C18 HPLC Column (5 μm, 10 × 250 mm, Thermo Fisher) and 5% acetonitrile as the mobile phase. The standard F6P, AcCoA, OAA, and sucrose (Sigma) were diluted to 1,000, 500, 100, 10, and 1 mg/L and analyzed similarly to generate the standard curve.

Li H., Pham N.N., Shen C.R., Chang C.W., Tu Y., Chang Y.H., Tu J., Nguyen M.T, & Hu Y.C. (2022). Combinatorial CRISPR Interference Library for Enhancing 2,3-BDO Production and Elucidating Key Genes in Cyanobacteria. Frontiers in Bioengineering and Biotechnology, 10, 913820.

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UPLC-APCI-FT-MS Analysis of Metabolites

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LC-APCI-FT-MS was performed using a Hypersil BDS C18 HPLC column (100 x 2.1 mm, 3 μm; Thermo Fisher Scientific) mounted on an Accela UPLC system (Thermo Electron Corporation). The LC system was coupled to a LTQ FT Ultra (Thermo Electron Corporation) via an atmospheric pressure chemical ionization (APCI) system operated in positive mode. The following gradient was run using water/acetonitrile (99:1, v/v) (solvent A) and acetonitrile/water (99:1, v/v) (solvent B): time 0 min, 50% B; 19 min, 99.5% B; 25 min, 99.5% B. The injection volume was 10 μL, flow rate 400 μL/min, and column temperature 40°C. Positive ionization using the APCI source was obtained with the following parameter values: capillary temperature 200°C, APCI probe 350°C, sheath gas 40 (arbitrary units), aux. gas 5 (arbitrary units), spray voltage 6.0°kV, discharge current 5 μA, capillary voltage 4.0 V, and tube lens 70 V. Full MS spectra between m/z 250-1400 were recorded at a resolution of 100,000. Full MS spectra were interchanged with a dependent MS 2 scan event in which the most abundant ion in the previous full MS scan was fragmented, two dependent MS 3 scan events in which the two most abundant daughter ions were fragmented, and a dependent MS 4 scan event in which the most abundant granddaughter ion of the first MS 3 scan event was fragmented. The collision energy was set at 35%.

Bicalho K.U., Santoni M.M., Arendt P., Zanelli C.F., Furlan M., Goossens A, & Pollier J. (2019). CYP712K4 Catalyzes the C-29 Oxidation of Friedelin in the Maytenus ilicifolia Quinone Methide Triterpenoid Biosynthesis Pathway. Plant & cell physiology, 60(11).

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Quantifying Anthocyanins in Flower Petals

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For the determination of the anthocyanin content, 10 g of shock-frozen petals were pulverized and mixed with 35 ml 2 M methanolic hydrochloric acid. After shaking for 12 hours at 4°C in an overhead rotator, the suspension was centrifuged for 10 minutes at 19200×g. 10–140 µl of the supernatant was adjusted with 2 M methanolic hydrochloric acid to a final volume of 1000 µl. The absorption at 520 nm was determined on a DU-65 spectrophotometer (Beckman Instruments). The anthocyanin content was calculated as pelargonidin equivalent using a calibration curve obtained with commercially available pelargonidin chloride (Roth, Germany). For acidic hydrolysis of anthocyanins, 20 µl methanolic hydrochloric acid extract were mixed with 180 µl of 4 N HCL and incubated for 60 minutes at 90°C. After cooling for 10 minutes the mixture was centrifuged for 10 minutes at 10000×g. The supernatant was adjusted to 200 µl with 4 N HCL and aliquots were used for HPLC analysis [32] (link) using a Perkin Elmer Series 200 HPLC system equipped with a Perkin Elmer Series 200 diode array detector and Total Chrom Navigator, version 6.3.1 (Perkin Elmer Inc). The column was a BDS Hypersil C18 HPLC column, 5 µm, 250×4.6 mm (Thermo Scientific).

Miosic S., Thill J., Milosevic M., Gosch C., Pober S., Molitor C., Ejaz S., Rompel A., Stich K, & Halbwirth H. (2014). Dihydroflavonol 4-Reductase Genes Encode Enzymes with Contrasting Substrate Specificity and Show Divergent Gene Expression Profiles in Fragaria Species. PLoS ONE, 9(11), e112707.

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HPLC Separation of Long-Chain Fatty Acids

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Example 2

Sample injection was performed with an Agilent Technologies G1367B Autosampler.

The autosampler system automatically injected an aliquot of the above prepared reconstituted samples into a Thermo Scientific BDS Hypersil C18 HPLC column (3 μm particle size, 100×2.1 mm, from Thermo Scientific). An HPLC gradient was applied to the analytical column, to separate VLCFA and BCFA from other components in the sample. Mobile phase A was 20 mM ammonium acetate and mobile phase B was 82% acetonitrile in methanol. The HPLC gradient started with an 82% solvent B which was ramped to 90% in approximately 1 minute, then ramped up to 95% for another minute, and held at that percentage for approximately 36 seconds, before being ramped back down to 90% over the next one minute and 18 seconds, and then down to 82% over the next 24 seconds. Column flow rate during solvent application was about 0.85 mL/min. Pristanic acid, phytanic acid, docosanoic acid, tetracosanoic acid, and hexacosanoic acid were observed to elute off the column at approximately 1.43 minutes into the gradient profile.

US10242852B2. Mass spectrometric determination of fatty acids (2019-03-26). Quest Diagnostics Investments Incorporated [US]. Inventors: Scott Goldman [US].

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HPLC Quantification of DPV Compound

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A Waters HPLC system (Elstree, UK) consisting of a 1525 binary HPLC pump, a 717 plus autosampler, an in-line degasser unit, a 1500 series column heater, a 2487 dual wavelength absorbance detector, and a 2998 photodiode array detector was used for all analyses. Release and solubility samples (10 µL), appropriately diluted, were injected onto a Thermo Scientific BDS Hypersil™ C18 HPLC column (150 mm × 4.6 mm, 3 µm particle size) maintained at 25 °C and fitted with a guard column. Isocratic elution was performed using a mobile phase comprising 60% HPLC-grade acetonitrile and 40% phosphate buffer (pH 3.0) and a total flow rate of 1.5 mL/min. The run time was 6 min and DPV was detected at 287 nm after ~2.85 min. DPV amounts were determined using a weighted calibration curve generated from three separate sets of dilutions performed independently in IPA/water (Supplementary Information, Fig. S1 and Table S1).

Murphy D.J., Lim D., Armstrong R., McCoy C.F., Bashi Y.H., Boyd P., Derrick T., Spence P., Devlin B, & Malcolm R.K. (2021). Refining the in vitro release test method for a dapivirine-releasing vaginal ring to match in vivo performance. Drug Delivery and Translational Research, 13(8), 2072-2082.

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HPLC Analysis of Drug Release

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All in vitro release and content samples were analysed on a Waters HPLC system (Waters Limited, Elstree, UK) consisting of a 1525 Binary HPLC pump, a 717 Plus Autosampler and a 2487 Absorbance Detector. Samples (25 μL) were injected onto a Thermo Scientific BDS Hypersil™ C18 HPLC column (150 mm ⨉ 4.6 mm, 3 μm particle size) fitted with a guard column. The column was held at 25 °C and isocratic elution was performed using a mobile phase comprising 58% HPLC-grade acetonitrile and 42% pH 2.4 phosphate buffer (prepared using 7.7 mM pH 3.0 phosphate buffer modified by addition of 2.2 mL of 85% H₃PO₄ and 1 mL of 10 M NaOH), a flow rate of 1.2 mL/min and a run time of 5 min. DPV was detected at 210 nm after approximately 2.5 min and LNG at 240 nm after approximately 3.5 min (Dallal Bashi et al., 2019 (link)).

Murphy D.J., Dallal Bashi Y.H., McCoy C.F., Boyd P., Brown L., Martin F., McMullen N., Kleinbeck K., Dangi B., Spence P., Hansraj B., Devlin B, & Malcolm R.K. (2022). In vitro drug release, mechanical performance and stability testing of a custom silicone elastomer vaginal ring releasing dapivirine and levonorgestrel. International Journal of Pharmaceutics: X, 4, 100112.

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Quantification of DPV and LNG via HPLC

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Quantification of DPV and LNG was performed on a Waters HPLC system (Waters Limited, Elstree, UK) consisting of a 1525 Binary HPLC pump with an in-line degasser AF unit, column heater, 717 Plus Autosampler and a 2487 dual wavelength absorbance detector. Samples (25 μL) were injected onto a Thermo Scientific BDS Hypersil™ C18 HPLC column (150 mm × 4.6 mm, 3 μm particle size) fitted with a guard column. The column was held at 25 °C and isocratic elution was performed using a mobile phase of 58% HPLC-grade acetonitrile and 42% pH 2.4 phosphate buffer (prepared using 7.7 mM, pH 3 phosphate buffer modified by addition of 2.2 mL of 85% phosphoric acid and 1.0 mL of 10 M sodium hydroxide), a flow rate of 1.2 mL/min and a run time of 5 min. DPV was detected at 210 nm at ~2.7 min and LNG at 240 nm at ~3.7 min (Dallal Bashi et al., 2019 (link)).

Dallal Bashi Y.H., Murphy D.J., McCoy C.F., Boyd P., Brown L., Kihara M., Martin F., McMullen N., Kleinbeck K., Dangi B., Spence P., Hansraj B., Devlin B, & Malcolm R.K. (2021). Silicone elastomer formulations for improved performance of a multipurpose vaginal ring releasing dapivirine and levonorgestrel. International Journal of Pharmaceutics: X, 3, 100091.

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HPLC Analysis of DPV Release

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A Waters HPLC system (Elstree, UK) consisting of; a 1525 binary HPLC pump, a 717 plus autosampler, an in-line degasser unit, a 1500 series column heater, a 2487 dual wavelength absorbance detector and a 2998 photodiode array detector was used for all analysis.
In vitro release 25 µL of sample was injected on to a Thermo Scientific BDS Hypersil C18 HPLC column (150 x 4.6 mm, 3 µm particle size) fitted with a guard column. The column was held at 45˚C and isocratic elution performed using a mobile phase of 45% acetonitrile and 55% phosphate buffer (pH 3.0, 7.7 mM) with a total flow rate of 1.2 mL/min and a run time of 8 min. DPV was detected at 240 nm after approximately 6.1 min.

Murphy D.J., McCoy C.F., Boyd P., Derrick T., Spence P., Devlin B, & Malcolm R.K. (2019). Drug stability and product performance characteristics of a dapivirine-releasing vaginal ring under simulated real-world conditions. International journal of pharmaceutics, 565.

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HPLC Analysis of DPV Content and Release

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Samples for DPV content analysis in rings were analysed on a Waters HPLC system (Waters Corporation, Dublin, Ireland) consisting of a 1525 Binary HPLC pump with an inline degasser AF unit, 1500 column heater, 717 Plus Autosampler and a 2487 dual wavelength absorbance detector. 10 µL of each content sample was injected onto a Kromasil C18 HPLC column (150 mm x 4.6 mm, 5 µm particle size). Column temperature was maintained at 25 °C and isocratic elution was performed using a mobile phase of 75% HPLC-grade methanol and 25% water with a flow rate of 0.75 mL/min and a run time of 15 min. DPV was detected at 257 nm after approximately 10.8 min.
In vitro release samples (25 µL) were injected onto a Thermo Scientific BDS Hypersil™ C18 HPLC column (150 mm x 4.6 mm, 3 µm particle size) fitted with a guard column. The column was held at 45 °C and isocratic elution was performed using a mobile phase of 45% HPLC-grade acetonitrile and 55% phosphate buffer (pH 3.0; 7.7 mM) with a run time of 8 min. DPV was detected at 240 nm after 6.1 min.

McCoy C.F., Murphy D.J., Boyd P., Derrick T., Spence P., Devlin B, & Malcolm R.K. (2017). Packing Polymorphism of Dapivirine and Its Impact on the Performance of a Dapivirine-Releasing Silicone Elastomer Vaginal Ring. Journal of pharmaceutical sciences, 106(8).

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