YMC USER'S MANUAL

Column Care and Use Instructions

CHIRAL ART Amylose-SA/Cellulose-SB
Immobilized type1.

Introduction
Thank you for purchasing a YMC high-performance liquid chromatography (HPLC) column. The CHIRAL ART Amylose-SA andCellulose-SB columns are designed for separating optical isomers (in normal/ reversed phase mode). The immobilized chiralselectors, polysaccharide derivatives, provide high compatibility with wide range of organic solvents, and superior separation andselectivity. With its advantages, CHIRAL ART Amylose-SA and Cellulose-SB columns are suitable for separating a variety of chiralcompounds.
CHIRAL ART Amylose-SA and Cellulose-SB columns, which are manufactured under highly controlled conditions, must pass aseries of stringent tests before being accepted for shipment. (Please refer to the column inspection report). To ensure optimal
performance and durability of the column, please read these instructions carefully before using the columns.

2.

Specifications
Item

CHIRAL ART Amylose-SA

CHIRAL ART Cellulose-SB
3, 5, 10, 20 µm

Particle size

Amylose tris(3,5-dimethylphenylcarbamate)

Chiral selectorType

Cellulose tris(3,5-dimethylphenylcarbamate)

Immobilized type

Separation mode

Normal Phase/ Reversed Phase 1

Shipping solvent 1

n-hexane/2-propanol (90/10)

Temp. range (℃)

0 – 40℃ (Max. 25℃ in pH7 – 9)Usable pH range

pH 2 – 9

Pressure limit

2

Recommended flow rate

30 MPa3

4.6 mm I.D.:
10 mm I.D. :0.5 – 1.0 mL/min (Max. flow rate： 3.0 mL/min)
2.5 – 5.0 mL/min (Max. flow rate： 15 mL/min)

1: After use in normal phase; if you intend to store the column for a long time, replace the mobile phase in the column with shippingsolvent.
The initial use in reversed phase (aqueous); replace the shipping solvent with ethanol or 2-propanol before replace with themobile phase for separation.
After use in reversed phase; the mobile phase should be replaced with ethanol before storing in the shipping solvent. For themobile phase containing buffer salts/additives, the replacement process should be carried out with caution to prevent theprecipitation of salt when replacing.
2: Avoid using a column repeatedly near the pressure limit or abrupt change in the pressure in order to prevent from shortening thecolumn lifetime. The recommended operational pressure of the column is 25 MPa or less.
3: Adjust flow rate according to the recommendation in the table above to obtain the optimum results under the application. The

repeated use at or near the upper limit of flow rate can reduce the column life time. When using column dimensions other thanlisted, adjust flow rate according to the cross-section area of the column.
2, 3: Pressure changes depending on column length, temperature, types of organic solvent etc. If pressure exceeds the upper limit,

reduce flow rate to below the lower rate of recommended range.

(UM150428AE) 1/3

YMC USER'S MANUAL

3.

Column installation
・ The column endfitting is Waters style connection.
・ Tubing must have flat ends and must bottom out in the column endfitting. Tubing must be connected to the column correctly toavoid creating a void between the column frit and tubing, which can cause a leak and result in poor column performance (e.g.
peak tailing, loss of theoretical plate number).
・ The correct direction of the solvent flow is indicated by an arrow on the column identification label.
・ Do not disconnect a column from the LC system before the pressure drops to zero.

4.

Mobile phase and sample solvent
・ CHIRAL ART are suitable for any mobile phase commonly used in HPLC columns, from aqueous to non-aqueous solvents.
The column can be used on both normal phase and reversed phase (aqueous mobile phase). We recommend that the column
is dedicated to either phase. Frequent switch of modes between phases can cause extensive damage to the column. Pleaserefer to the recommendation of the solvents in the table below.
・ After replacing among solvents with difference in contained organic solvent or in pH value, sufficiently equilibrate the column
with the mobile phase. Depending on the mobile phase composition, it maybe take over 100 (one hundred) column volumes
running.
・ Make sure of miscibility among the organic solvents. When switching the mobile phase from alkane/alcohol mobile phase topolar organic solvents (methanol, acetonitrile etc), flush the column with compatible solvent such as ethanol or 2-propanol for atleast 10 (ten) column volumes beforehand. After that, sufficiently equilibrate the column with the mobile phase.
・ Be careful of the solvent resistance of HPLC system and PEEK tubing when using them on normal phase mode.
・ When a target compounds is ionic, addition of modifier listed below can improve peak shape and/or separation reproducibility.
High concentrations of modifiers can result in reducing column lifetime. Add/reduce the modifiers according to the notes in thetable.
・ Whenever possible, the sample should be dissolved in the same composition as the initial mobile phase. Using a stronger
solvent than mobile phase for sample dissolution may result in broad peaks and reducing the separability and reproducibility. Inaddition, before injection, please check the miscibility of the sample solvent and mobile phase in order to prevent the sample
from precipitating on injection.
・ In order to avoid blockage which can cause pressure increase, the sample solution should be filtered through a membrane filter(0.2 μm or smaller porosity).
【Recommended solvents for Normal phase】
Acidic compounds
Organic solvents
Modifiers

Basic compounds

Non-ionic compounds

alkane (n-hexane or n-heptane), alcohols(methanol, ethanol, 2-propanol), acetonitrile, ethyl acetate,tetrahydrofuran (THF), dichloromethane, chloroform, methyl tert-butyl ether (MTBE)
0.1%(Upper limit 0.5%)
trifluoroacetic acid (TFA),
acetic acid, formic acid, etc

Composition ratio

0.1%(Upper limit 0.5%)
diethylamine (DEA), butylamine,ethanolamine, etc

None

Any ratio (those should be miscible)【Recommended solvents for Reversed phase】
Acidic compounds
Organic solvents
Aqueous phase(Modifiers)
Composition ratio

Basic compounds

Non-ionic compounds

Acetonitrile, methanol, ethanol, 2-propanol, THF, etc
0.1%phosphoric acid,
0.1%formic acid,50 – 100 mM phosphate buffer(pH2.0 – 3.5), etc

20 mM NH4HCO3-DEAbuffer(pH9.0), etc

Water

Organic solvent/ aqueous solution (10/90 – 100/0)(UM150428AE) 2/3

YMC USER'S MANUAL

5.

Column cleaning (general method)
The column needs to be replaced when the cleaning methods below do not regenerate the column performance. To extend thecolumn lifetime, especially for samples containing large amount of impurities, we recommend a sample pretreatment conductedcarefully prior to introducing the sample to the column and /or a guard column to use with.
【Normal phase】
・ Flush the column with the one of the organic solvents, which has the highest polarity among the composition of mobile phasewith increased concentration (for example, for alkane/alcohol mobile phase, concentration of alcohol should be increased) towash out the residual substances strongly retained on the column. For further cleaning, flush with 100% ethanol.
・ If 100% ethanol does not improve the column performance, flush10 (ten) column volumes (30 (thirty) column volumes: whenmobile phase contains modifiers) of N, N-dimethylformamide (DMF) on Amylose-SA column and of ethyl acetate on
Cellulose-SB column. There is a possibility that the column performance is restored after storing the column with thecorresponding solvent for several days.
・ When the mobile phase is containing acid or amine, replace with solvent containing neither of them (at the same ratio as themobile phase), then wash as above procedure. Do not store the column with solvent containing modifiers even for a shortperiod of time.
【Reversed phase】
・ Flush the column with solution containing a higher ratio of organic solvent for washing out the compounds that have a great
capacity for retention in the column after using mobile phases not containing buffer salts/additives. Usable concentration oforganic solvent is up to 100%.
・ When using mobile phase containing buffer salts/additives, first replace with a water/organic solution containing no buffersalts/additives (A ratio of water to organic solvent should be set at the same proportions as a mobile phase). Then flush thecolumn in accordance with the method described above.

YMC Karasuma-Gojo Bldg., 284 Daigo-cho, Karasuma Nishiiru Gojo-dori, Shimogyo-ku,
Kyoto 600-8106, Japan
TEL +81-75-342-4515 FAX +81-75-342-4550
URL:www.ymc.co.jp
E-Mail:sales@ymc.co.jp(UM150428AE) 3/3