Spiral Countercurrent Chromatography Studies Using the Spiral Disk Assembly

Knight, Martha; Finn, Thomas M.

doi:10.1080/10826070903245714

Cited by 11 publications

(21 citation statements)

References 11 publications

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“…2 . The STS rotors are manufactured under a license from the National Institutes of Health [17–18]. The STS rotor has circular channels holding tubing that is wound in a spiral path to the center, skipping 3 concentric channels.…”

Section: Methodsmentioning

confidence: 99%

Isolation of sutherlandins A, B, C and D from Sutherlandia frutescens (L.) R. Br. by counter-current chromatography using spiral tubing support rotors

Chen¹,

Folk

Lazo-Portugal³

et al. 2017

Journal of Chromatography A

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Spiral countercurrent-chromatography has great potential for improving the capacity and efficiency of purification of secondary metabolites, and here we describe applications useful for the isolation of flavonoids from the widely used South African medicinal plant, Sutherlandia frutescens (L.) R. Br. In the spiral tubing support rotor, STS-4 for high-speed counter-current chromatography, several polar butanol aqueous solvent systems were selected using a Log K plot, and the novel flavonol glycosides (sutherlandins A-D) were well separated by the optimized solvent system (ethyl acetate; n-butanol; acetic acid; water; 5:1:0.3:6 by vol.). The yield of purified flavonoids from 0.9 g extract varied from 8.6 mg to 54 mg of the sutherlandins for a total of 85.3 mg. The same extract was fractionated in the new STS-12 rotor of the same outside dimensions but with more radial channels forming 12 loops of the tubing instead of 4. The rotor holds more layers and increased length of tubing. From 0.9 g extract the STS-12 rotor yielded more recovery of 110.4 mg total with amounts varying from 11.2 mg to 64 mg of the sutherlandins. and apparent increased separation efficiency as noted by less volume of each fraction peak. Thus from 1-g amounts of extract, good recovery of the flavonoids was achieved in the butanol aqueous solvent system

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Section: Methodsmentioning

confidence: 99%

Isolation of sutherlandins A, B, C and D from Sutherlandia frutescens (L.) R. Br. by counter-current chromatography using spiral tubing support rotors

Chen¹,

Folk

Lazo-Portugal³

et al. 2017

Journal of Chromatography A

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“…A Conway Centri Chrom (Williamsville, NY, USA) planetary centrifuge [4] was mounted with a STS rotor (17-cm OD, CC Biotech LLC, Rockville, MD, USA) [5] (Fig. 2).…”

Section: Methodsmentioning

confidence: 99%

“…The lower phase was pumped at a flow of 1 mL/min and 4 or 5-min fractions were collected. The elution mode was L-i-H, meaning the mobile lower phase was introduced in the inner (top) entry which is head to tail in the CW direction of rotation used [4]. After elution of mobile phase, around 200 ml, the rotation was ceased and contents pushed out with He gas pressure.…”

Section: Methodsmentioning

confidence: 99%

“…We found that in these new rotors, the n-butanol/0.1M K 2 HPO 4 , KH 2 PO 4 (1:1) solvent system had a S F of 70% and sec -butanol/0.5% aq. trifluoroacetic acid (1:1) had a S F of 66% [4,5] compared to 40% or less in the multi-layer coil. Higher S F results in greater separation efficiency.…”

Section: Introductionmentioning

confidence: 99%

“…Here we investigated the addition of n -butanol to an organic-aqueous solvent system to modify the partitioning and achieve separation of various triterpenoid glycosides in an extract. Previously, we had studied the operation of the spiral disk rotor in some detail with non-polar to polar organic-aqueous solvent systems [4] and later studied the STS rotor and its applicability to small and larger molecules using the polar organic-aqueous solvent systems. Here we are applying this rotor for the first time to small molecule natural products separation.…”

Section: Introductionmentioning

confidence: 99%

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Isolation of the Predominant Cycloartane Glycoside, Sutherlandioside B, from Sutherlandia frutescens (L.) R.Br. by Spiral Countercurrent Chromatography

Brownstein

Rottinghaus²,

Knight

et al. 2014

Journal of Liquid Chromatography & Related Technologies

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The introduction of spiral countercurrent chromatography in the last few years using new separation columns such as the spiral tubing support rotor has enabled the application of more polar volatile solvent systems for natural products separation. This method can be applied to water soluble compounds and their metabolites. We have used spiral countercurrent chromatography with the spiral tubing support rotor to fractionate n-butanol extracts of an African plant Sutherlandia frutescens and have determined conditions by which the predominant cycloartane glycoside (sutherlandioside B) can be purified in good yield. A solvent system of ethyl acetate, methanol, and water was modified by adding n-butanol to separate sutherlandioside B from other compounds. With the optimal amount of n-butanol in the two-phase solvent system with the lower aqueous phase mobile, the target compound was eluted well separated from the other components. The purity of sutherlandioside B was determined by high performance liquid chromatography/mass spectrometry analysis and the yield compares favorably with the content in bulk material.

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Spiral counter-current chromatography of small molecules, peptides and proteins using the spiral tubing support rotor

Knight¹,

Finn²,

Zehmer³

et al. 2011

Journal of Chromatography A

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An important advance in countercurrent chromatography (CCC) carried out in open flow-tubing coils, rotated in planetary centrifuges, is the new design to spread out the tubing in spirals. More spacing between the tubing was found to significantly increase the stationary phase retention, such that now all types of two-phase solvent systems can be used for liquid-liquid partition chromatography in the J-type planetary centrifuges. A spiral tubing support (STS) frame with circular channels was constructed by laser sintering technology into which FEP tubing was placed in 4 spiral loops per layer from the bottom to the top and a cover affixed allowing the tubing to connect to flow-tubing of the planetary centrifuge. The rotor was mounted and run in a P.C. Inc. type instrument. Examples of compounds of molecular weights ranging from <300 to approximately 15,000 were chromatographed in appropriate two-phase solvent systems to assess the capability for separation and purification. A mixture of small molecules including aspirin was completely separated in hexane-ethyl acetate-methanol-water. Synthetic peptides including a very hydrophobic peptide were each purified to a very high purity level in a sec-butanol solvent system. In the STS rotor high stationary phase retention was possible with the aqueous sec-butanol solvent system at a normal flow rate. Finally, the two-phase aqueous polyethylene glycol-potassium phosphate solvent system was applied to separate a protein from a lysate of an E. coli expression system. These experiments demonstrate the versatility of spiral CCC using the STS rotor.

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Spiral Countercurrent Chromatography Studies Using the Spiral Disk Assembly

Cited by 11 publications

References 11 publications

Isolation of sutherlandins A, B, C and D from Sutherlandia frutescens (L.) R. Br. by counter-current chromatography using spiral tubing support rotors

Isolation of sutherlandins A, B, C and D from Sutherlandia frutescens (L.) R. Br. by counter-current chromatography using spiral tubing support rotors

Isolation of the Predominant Cycloartane Glycoside, Sutherlandioside B, from Sutherlandia frutescens (L.) R.Br. by Spiral Countercurrent Chromatography

Spiral counter-current chromatography of small molecules, peptides and proteins using the spiral tubing support rotor

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