For the purification of unidentified GOS, the reaction was carried out as described above and stopped after 22 h by heating the solution for 10 min at 96 ºC to inactivate the biocatalyst. The reaction mixture was filtered using 0.45 µm paper filters and purified by semipreparative hydrophilic interaction chromatography (HPLC-HILIC). A quaternary pump (Delta 600, Waters) coupled to a Lichrosorb-NH2 column (5 m, 10 x 250 mm, Merck) was used. The column temperature was kept at 25 ºC. The acetonitrile/water 75/25 (v/v) mobile phase was conditioned with helium and used as at a flow-rate of 6.25 mL/min for 43 min. Peaks were detected by using an evaporative light-scattering detector DDL-31 (Eurosep) equilibrated at 60 C. A three-way flow splitter (model Accurate, Dionex) and a fraction collector II (Waters) were employed. The fractions containing the main peaks were collected and the solvent was removed by rotary evaporation.
Fraction collector 2
Manufactured by Waters Corporation
Sourced in Germany
The Fraction Collector II is a versatile laboratory instrument designed to automatically collect fractions of liquid samples. It features a compact and user-friendly design, allowing for efficient and reliable fraction collection during various analytical processes.
Automatically generated - may contain errors
Lab products found in correlation
Silica gel, by Qingdao Haiyang Chemical (3 mentions)
Lichrosorb nh2 column, by Merck Group (1 mentions)
Delta 600, by Waters Corporation (1 mentions)
Waters 2996 photodiode array detector, by Waters Corporation (1 mentions)
Dip 370, by Jasco (1 mentions)
Sepa 300, by Jasco (1 mentions)
G6230 tof ms, by Agilent Technologies (1 mentions)
Am 400, by Bruker (1 mentions)
2401a spectrophotometer, by Shimadzu (1 mentions)
Drx 500, by Bruker (1 mentions)
Avance 3 600, by Bruker (1 mentions)
3 protocols using fraction collector 2
1
Purification of Unidentified Glycosidic Compounds
2
Analytical Techniques for Natural Product Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Optical rotations were carried out using a Horiba SEPA-300 polarimeter and JASCO DIP-370 digital polarimeter. UV spectra were recorded on Shimadzu 2401Aspectrophotometer. IR Spectra were obtained on Brucker Tensor 27 infrared spectrophotometer with KBr pellets. 1H, 13C and 2D NMR spectral data were measured on a Bruker Avance III-600, DRX-500, and AM-400 MHz spectrometers with SiMe4 as an internal standard. HRESIMS data were recorded on an Agilent G6230 TOF MS. Column chromatography (CC) was performed with silica gel (200–300 mesh, Qing-dao Haiyang Chemical Co., Ltd., Qingdao, China). RP-18 silica gel (20–45 μm, Fuji Silysia Chemical Ltd., Japan). Fractions were monitored by TLC on silica gel plates (GF254, Qingdao Haiyang Chemical Co., Ltd.) and spots visualized with Dragendorff’s reagent spray. MPLC was employed using a Buchi pump system coupled with RP-18 silica gel packed glass columns(15 × 230 and 26 × 460 mm, respectively). HPLC system was carried out on a Waters HPLC system (Waters 1525E pumps, Waters 2996 photodiode array detector, Waters fraction collector II) using a analytical semi-preparative or preparative Sunfire C18 column (4.6 × 150, 10 × 150, and 19 × 250 mm, respectively).
3
Elastin Fractionation by Reversed-Phase HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Elastin was dispersed in 50 mM Tris HCl, pH 8.5, at a concentration of 5 mg mL -1 . PE was added to a final enzyme-tosubstrate ratio of 1:100 (w/w) and the samples were subsequently incubated at 37°C for 24 h. The digestion was stopped by adding TFA to a final concentration of 0.5% (v/v), and the samples were stored at -26°C prior to further analysis. Fractionation of elastin digests was performed using an Agilent 1100 (Waldbronn, Germany) coupled to a Fraction Collector II (Waters, Manchester, UK) at a constant flow rate of 200 μL min -1 . For chromatographic separation, the injected sample was loaded onto a Reprosil-Pur 120 column (C18, 3 μm, 2 mm i.d. × 150 mm; Dr. Maisch GmbH, Ammerbuch-Entringen, Germany) and eluted using a solvent system of solvent A (0.1% FA in H 2 O) and solvent B (0.1% FA in ACN/H 2 O 80:20 (v/v)) by applying a linear binary gradient: 10%-40% B in 40 min, to 90% B in the next 5 min, followed by maintenance at 90% B for 10 min, and then from 90% to 10% B in 5 min. Fractions were collected every 30 s.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!