Method of purification and purified products

A method of packing a chromatography column in column chromatography comprising:

• adding a stationary phase to a column, and
• allowing a slurry comprising the stationary phase to settle in the column for a standing time, wherein the standing time is about 10 minutes or longer, and wherein a mobile phase is not discharged during the standing time, wherein the stationary phase is a reverse phase silica stationary phase, and the compound to be purified by column chromatography is selected from fatty acids derived from fish oil or esterified derivatives thereof, proteins and nucleic acids, and wherein chromatography comprises elution with a mobile phase comprising one or more of water, methanol, ethanol, or acetonitrile, and
• wherein the addition of the stationary phase is conducted by:
• a) adding the slurry to the column after the slurry is formed by premixing the stationary phase with a mobile phase;
• or
• b) adding a mobile phase to the column then adding the stationary phase to the mobile phase in the column.

A method of increasing theoretical plate number (N) in a column chromatography comprising:

• adding a stationary phase to a column, and
• allowing a slurry comprising the stationary phase to settle in the column for a standing time, wherein a mobile phase is not discharged during the standing time,
• performing the column chromatography using the column after the slurry is settled,
• performing a gas chromatography of a resulting compound obtained by the column chromatography, and
• calculating the theoretical plate number (N) according to equation (1) below,
• N
• =
• (
• t
• R
• σ
• )
• 2
• ,
• (
• 1
• )
• wherein σ is a standard deviation and tR is a retention time,
• wherein the increase in Nis relative to a column chromatography without standing time or wherein the increase in N is relative to a slurry that settles for a reduced standing time that is less than the standing time, wherein the stationary phase is a reverse phase silica stationary phase, and the compound to be purified by column chromatography is selected from fatty acids derived from fish oil or esterified derivatives thereof, proteins and nucleic acids, and wherein chromatography comprises elution with a mobile phase comprising one or more of water, methanol, ethanol, or acetonitrile,
• wherein the standing time is about 10 minutes or longer, and
• wherein the addition of the stationary phase is conducted by:
• a) adding the slurry to the column after the slurry is formed by premixing stationary phase with a mobile phase;
• or
• b) adding a mobile phase to the column then adding the stationary phase to the mobile phase.

A method of improving symmetry factor of a chromatographic peak in column chromatography comprising:

• adding a stationary phase to a column,
• allowing a slurry to settle in the column for a standing time, wherein a mobile phase is not discharged during the standing time,
• performing the column chromatography using the column after the slurry is settled,
• performing a gas chromatography of a resulting compound obtained by the column chromatography, and
• calculating a symmetry factor according to equation (2) below,
• S
• =
• W
• 0.05
• h
• 2
• ⁢
• f
• ,
• (
• 2
• )
• wherein W0.05h is a peak width at 5% of a height of a chromatographic peak, and f is a peak width of a first half of the chromatographic peak measured as a horizontal distance from a point of a beginning of the chromatographic peak to an apex of a peak amplitude at 5% of the height of the chromatographic peak, and a resulting compound obtained by the column chromatography is selected from fatty acids derived from fish oil or esterified derivatives thereof, proteins and nucleic acids, and wherein chromatography comprises elution with a mobile phase comprising one or more of water, methanol, ethanol, or acetonitrile, and wherein the standing time is about 10 minutes or longer, and
• wherein the addition of the stationary phase is conducted by:
• a) adding the slurry to the column after the slurry is formed by premixing the stationary phase with a mobile phase;
• or
• b) adding a mobile phase to the column then adding the stationary phase to the mobile phase,
• wherein the improvement comprises decreasing the symmetry factor of the peak to a quantity ≥1, and wherein the decrease is relative to a symmetry factor when the column chromatography comprises a reduced standing time that is absent or less than the standing time and the symmetry factor is >1;
• or
• the improvement comprises increasing the symmetry factor of the peak to a quantity ≤1, and wherein the increase is relative to a symmetry factor when a the column chromatography comprises a reduced standing time that is absent or less than the standing time and the symmetry factor is <1, and wherein the slurry comprises a stationary phase that is a reverse phase silica stationary phase, and the compound to be purified by column chromatography is selected from fatty acids derived from fish oil or esterified derivatives thereof, proteins and nucleic acids, and wherein chromatography comprises elution with a mobile phase comprising one or more of water, methanol, ethanol, or acetonitrile.