7 Reversed-Phase Columns (ther than DS) Types and characteristics of reversed-phase columns -- 9, 9 YMC-Pack Pro C8, C--------------------------- 96 YMC-Pack C 8 -------------------------------------- 97 YMC-Pack C -------------------------------------- 97 YMC-Pack TMS ----------------------------------- 98 YMC-Pack Ph -------------------------------------- 98 YMC-Pack CN ------------------------------------- 99 YMC-Pack PRTEIN-RP ------------------------ 99 YMCbasic----------------------------------------- YMC Carotenoid -------------------------------- rdering Information --------------------,
7 Types and characteristics of reversed-phase columns YMC reversed-phase columns include a variety of columns other than DS, enabling column selection from a wide range of products to suit the sample characteristics. Elution behavior dependent on alkyl chain length In reversed-phase chromatography, retention due to hydrophobicity generally depends directly on the carbon number of the stationary phase. The degree of retention due to hydrophobicity of the stationary phase can generally be listed in descending order by column type as DS>C8>C>TMS. Stationary phases with low hydrophobicity can be used effectively to reduce analysis time for samples having too strong of a retention on DS. Stationary phases with low hydrophobicity are also useful for samples that are slightly soluble in organic solvents and need to be analyzed with mobile phase containing a low concentration of organic solvents. Elution behavior affected by other factors Phenyl, PFP, and CN have available π electrons derived from their functional groups. Phenyl, PFP, and CN sometimes show different separation characteristics from stationary phases that are chemically bonded with straight alkyl chains. Since CN has medium-polar functional groups, it can be used in both normal-phase and reversed-phase separation modes, depending on the mobile phase used. Types of reversed-phase columns (I) DS -C 8 H 7 Retention due to hydrophobicity C8 -C 8 H 7 High C -C H 9 TMS -CH Types of reversed-phase columns (II) Phenyl (Ph) Types and characteristics of reversed-phase columns (other than DS) YMC-Triart Product name C8 Pore size (Å) Particle size (μm) C% Silanol treatment Usable ph range Yes Phenyl.9,, 7. ~. 7. ~. PFP No. ~ 8. Meteoric Core C8 8.7 Yes. ~ 9. PFP CN F F F F F -(CH ) -CN Characteristics Low π electrons available π electrons available π electrons available Can be used also in normal-phase Versatile hybrid silica based C8 column Ideal for separations of isomers or structural analogs Versatile hybrid silica based Phenyl column Ideal for separations of aromatic compounds or compounds having long conjugated system Versatile hybrid silica based PFP column Ideal for separations of polar compounds or isomers Core-Shell type C8 Ultra fast analysis and excellent resolution Pages 6 6 6 7~7 Pro series Pro C8, Yes Processed with advanced endcapping technology Superior separation of basic compounds 96 Pro C, 7 Processed with advanced endcapping technology Different selectivity from DS YMC-Pack series C 8, 7,,, C,,, 7, Yes. ~ 7. Moderate hydrophobicity Useful for separation of proteins and peptides Lower hydrophobicity than DS and C8 Useful for separation of proteins and peptides TMS,, Reversed-phase packing material with the lowest hydrophobicity 98 Ph,, 9 Reversed-phase packing material with π electrons 98,, 7 Can be used in both normal-phase and CN 99 reversed-phase modes PRTEIN-RP. ~ 7. Useful for separation of proteins and peptides 99 YMCbasic, 7 Yes. ~ 7. Superior separation of basic compounds Useful for separation of proteins and peptides YMC Carotenoid,. ~ 7. Useful for carotenoids separation 97 97 9
Elution behavior dependent on alkyl chain length Pro C8 Pro C8 Pro C min. Toluene. Deoxycorticosterone acetate. Imipramine hydrochloride. Triphenylene. n-amylbenzene Column Eluent Flow rate Temperature Detection min min : X.6 mmi.d. : mm KHP-KHP (ph 6.9)/methanol (/7) :. ml/min : 7 C : UV at nm In this example, the retention behavior of a variety of compounds is shown to be dependent on the alkyl chain length of the stationary phase. Shorter alkyl chain lengths like C show reduced retention for neutral compounds due to the diminished hydrophobicity of the C stationary phase relative to longer alkyl chains phases like C8 and C8. The differences in the selectivity of stationary phases of different alkyl chain length is also illustrated for triphenylene, a planar molecule with restricted rotational movement. Triphenylene shows much shorter retention on C relative to C8 and C8 than would be expected on the basis of hydrophobicity of the stationary phase. Note the difference in elution order for triphenylene relative to imipramine and amylbenzene for this mixture on this series of stationary phases. Comparison of separation selectivity among YMC-Triart reversed-phase columns Separation selectivity of YMCTriart reversed-phase columns is compared on analysis of biologically active amines and their related compounds. Retention time of each compound is summarized by type of compounds. As shown, Triart PFP column shows strong retention of basic compounds (peak,, 6,, ). It is considered that basic compounds which has electron-donating characteristic and polarised PFP group/silanol group are strongly interacted, and as a result, Triart PFP shows longer retention time. Good resolution with long retention Good resolution with long retention Biologically active amines and their related compounds Column Eluent : μm, X. mmi.d. : A) mm formic acid B) methanol containing mm formic acid -%B (- min), %B (- min) Flow rate :. ml/min Temperature : C Detection : UV at 8 nm 9
7 YMC-Pack Pro C8, C Superior separation of basic compounds Excellent reproducibility Utilizes highly pure silica gel base C8 Pore size : Å Carbon content : % Usable ph range :.~7. USP L7 C Pore size : Å Carbon content : 7% Usable ph range :.~7. USP L6 Highly endcapped C8 and C reversed-phase columns YMC-Pack Pro C8 and C are highly appropriate for basic compounds since more advanced endcapping technology is used for processing of their residual silanol groups that are likely to affect quality. The YMC-Pack Pro C8 and C stationary phase surface hydrophobicity is lower than that of DS, making YMC-Pack Pro C8 and C useful for quick analysis of compounds that differ greatly in hydrophobicity. The separation behavior of hydrophilic compounds or planar compounds on YMC-Pack Pro C8 and C also differs from that on DS, making YMC-Pack Pro C8 and C useful for separating compounds in cases where separation optimization is difficult to achieve using DS. ptimization of separation using Pro C8 and Pro C Separation of antiarrhythmics Pro C8 Buffer/MeH (/6) Pro C8 Buffer/MeH (/6) 6 min Pro C8 Buffer / MeH (/6) 7 ptimization of mobile phase 6 7 min Pro C Buffer / MeH (/6) 6 7 6 7 min min. Phenytoin. Propranolol HCl. Quinidine. Lidocaine. Diltiazem HCl 6. Verapamil HCl 7. Nicardipine HCl Column : X.6 mmi.d. Eluent : mm KH P -K HP (ph 6.9)/methanol Flow rate :. ml/min Temperature : 7 C Detection : UV at nm Retention times of analytes on Pro C8 and Pro C tend to be shorter than those on C8. When alkyl chain lengths of packing material functional groups are shorter, hydrogen-bonding capacities tend to be greater; therefore, not only retention time, but also separation selectivity of Pro C8 and Pro C may differ from those of C8. Separation optimization is difficult to achieve for antiarrhythmics using Pro C8, even if the mobile phase is changed. In contrast, C8 and C can completely separate antiarrhythmics in a short time. As shown above, C8 and C may be useful in cases where separation optimization is difficult to achieve using C8. 96
YMC-Pack C 8 Stationary phase with lower hydrophobicity than DS Useful for separating samples with relatively high hydrophobicity Useful for separation of proteins and peptides Pore size :,, Å Carbon content : %, 7%, % Usable ph range :.~7. USP L7 Reversed-phase column with moderate hydrophobicity The hydrophobicity of YMC-Pack C 8 is moderate for a reversed-phase packing material. Retention times of samples on YMC-Pack C 8 tend to be shorter than those on DS stationary phase. The moderate hydrophobicity of YMC-Pack C 8 makes it useful for separating samples with relatively high hydrophobicity. Application (K9A) Anti-HIV nucleoside derivatives HH C NH N N N HN N N HH C HN N.... HH C CH HH C HN N CH ddc ddi dt N AZT min YMC-Pack C Stationary phase with low hydrophobicity Different separation characteristics from DS Useful for separation of proteins and peptides Column : YMC-Pack C 8 ( μm, Å) X.6 mmi.d. Eluent : methanol/ mm KH P (/6) Flow rate :. ml/min Temperature : 7 C Detection : UV at nm Pore size :,, Å Carbon content : 7%, %, % Usable ph range :.~7. USP L6 Reversed-phase column with shorter alkyl chain The YMC-Pack C stationary phase surface hydrophobicity is lower than that of both DS and C 8. Retention times of samples on YMC-Pack C therefore tend to be shorter than those on DS or C 8. Separation characteristics of YMC-Pack C also differ from those of DS. YMC-Pack C achieves better separation than DS for some types of samples. Application (T9A) 9 6 8 7,-DNPH derivatives of aldehydes and ketones. Formaldehyde,-DNPH. Acetaldehyde,-DNPH. Acetone,-DNPH. Acrolein,-DNPH. Propionaldehyde,,-DNPH 6. Crotonaldehyde,-DNPH 7. Methylethylketone,-DNPH 8. Isobutyraldehyde,-DNPH 9. Benzaldehyde,-DNPH. n-valeraldehyde,-dnph. p-tolualdehyde,-dnph. Capronaldehyde,-DNPH Column :YMC-Pack C ( μm, Å) X.6 mmi.d. Eluent : A) tetrahydrofuran/water (/9) B) acetonitrile %B (-7 min), -6%B (7-8 min, linear), %B (8-9 min), %B (9- min) Flow rate :. ml/min Temperature : C Detection : UV at 6 nm min 97
7 YMC-Pack TMS Stationary phase with the lowest hydrophobicity among reversed-phase packing materials Different separation characteristics from DS Pore size : Å Carbon content : % Usable ph range :.~7. USP L Reversed-phase column with the lowest hydrophobicity YMC-Pack TMS shows lower retention due to hydrophobic interaction than other packing materials, and it is useful for eluting highly hydrophobic compounds in a short time. In addition, it can sometimes achieve greater retention and better separation of hydrophilic compounds than other reversed-phase columns. Shorten analysis time using TMS YMC-Pack Pro C8 Isocratic 6%B min YMC-Pack Pro C8 min YMC-Pack Ph Reversed-phase column with π electrons Unique selectivity due to π-π interaction Useful in cases where separation optimization is difficult to achieve using DS Different selectivity from DS change elution conditions Gradient -9%B (- min), 9%B (- min) change column YMC-Pack TMS Isocratic %B min Soy isoflavones. Daidzin. Genistin. Daidzein. Genistein Column : X. mmi.d. Eluent : A) water/formic acid (/.) B) acetonitrile/water/formic acid (//.) Flow rate :. ml/min Temperature :7 C Detection : ESI positive mode TMS enables analysis time of highly hydrophobic compounds to shorten. Pore size : Å Carbon content : 9% Usable ph range :.~7. USP L YMC-Pack Ph has π electrons of phenyl group. YMC-Pack Ph shows different separation characteristics from alkyl-silica stationary phases including DS for separation of solutes such as aromatic compounds, since π-π interaction between the stationary phase and solutes, as well as hydrophobic interaction, contribute to the separation. Establishment of simple conditions using Ph YMC-Pack Pro C8 acetonitrile/water/tfa (/88/.) change column N6A min change mobile phase YMC-Pack Ph acetonitrile/water/tfa (/88/.) Catechins. (-)-Epigallocatechin. (+)-Catechin. (-)-Epicatechin. (-)-Epigallocatechin gallate. (-)-Epicatechin gallate Column : X.6 mmi.d. Flow rate :. ml/min Temperature :7 C Detection : UV at 8 nm YMC-Pack Pro C8 acetonitrile/ethyl acetate/water/tfa (9//88/.) N6A min Ph is suitable for separating catechins with simple mobile phase, whereas if using DS and optimizing analysis conditions, the mobile phase, addition with ethyl acetate, is complicated. N6A min 98
YMC-Pack CN Normal-phase and reversed-phase modes are selectable according to the purpose of analysis Low hydrophobicity Unique selectivity due to cyano group Pore size :, Å Carbon content : 7%, % Usable ph range :.~7. USP L Column can be used in both normal-phase and reversed-phase modes YMC-Pack CN can be used in both normal-phase and reversed-phase modes, since it has cyanopropyl group of medium polarity chemically bonded to the stationary phase. It can be used in normal phase mode with low-polarity mobile phase such as hexane. It can also be used in reversed-phase mode with highly-polar mobile phase such as methanol and water. The hydrophobicity of YMC- Pack CN is relatively low for a reversed-phase packing material, and it shows different selectivity from DS due to π electrons of the cyano groups. YMC-Pack CN is useful for shortening analysis time when retention time is too long with DS and useful in cases where separation optimization is difficult to achieve using DS. Application (S9E) Anti-hyperlipidemia drugs. CH CI CNHCHCH C CH Bezafibrate CH. CH CH CH CI C CCH ( CH ) C CH CH CH CH Clofibrate Gemfibrozil.. CH CI C C CCH CH Fenofibrate CH CH min Column : YMC-Pack CN ( μm, Å) X.6 mmi.d. Eluent : acetonitrile/methanol/water/acetic acid (//6/) Flow rate :. ml/min Temperature :7 C Detection : UV at 7 nm YMC-Pack PRTEIN-RP Improved recovery of proteins or peptides Improved durability when used with TFA solution Enables elution of high molecular weight proteins Reversed-phase column for separation of proteins or peptides Pore size : Å Carbon content : % Usable ph range :.~7. USP L6 YMC-Pack PRTEIN-RP is a reversed-phase column utilizing a silica gel base. It contains a stationary phase, specifically designed for separation of proteins or peptides. Problems that are associated with conventional reversed-phase columns with short alkyl chain lengths are minimized. Robust column lifetime and excellent recovery of hydrophobic proteins are typically possible with this phase. Improved durability when used with TFA solution.%tfa condition <Prior to flow> <After hours> % retention of diphenyl 9 8 7 6 YMC-Pack PRTEIN-RP (Peak split) C column, Company A C column, Company B Solvent flow in hours min min Theoretical plate number () =, Theoretical plate number () =,8. Uracil. Benzene. Naphthalene. Diphenyl <Flow conditions> Eluent : water/tfa (/.) Flow rate :. ml/min Temperature :ambient <Measurement conditions> Column : YMC-Pack PRTEIN-RP X.6 mmi.d. Eluent : acetonitrile/water (/6) Flow rate :. ml/min Temperature : C Detection : UV at nm,. AUFS Test results of the stability of stationary phase with.% aqueous TFA is shown adove. Retention of diphenyl on C columns manufactured by other companies greatly decreases as time passes. This is caused by cleavage of butyl groups from the packing material due to acid hydrolysis. Retention of diphenyl on PRTEIN-RP is shown to be stable after hours of mobile phase flow. 99
7 YMCbasic Superior separation of basic compounds Useful for separation of peptides Secondary interaction minimized as much as possible Pore size : Å Carbon content : 7% Usable ph range :.~7. USP L7 Column for separation of basic compounds YMCbasic is a reversed-phase silica based C8 column designed for separation of basic compounds, including pharmaceutical products. It is highly evaluated as a base-deactivated phase in Europe and the U.S. It offers superior separation of acidic compounds as well as basic compounds. It is suitable for separating peptides with molecular weights in the range of several thousands, such as insulin. Application (N67C) mau 6 Tryptic digest of BSA YMC Carotenoid * 6 min * undigested BSA Resolves polar and nonpolar geometric carotenoid isomers Column : YMCbasic ( μm) X. mmi.d. Eluent : A) water/tfa (/.) B) acetonitrile/tfa (/.) -%B (- min), -%B (- min), %B (-6 min) Flow rate :. ml/min Temperature :7 C Detection : UV at nm Separates carotenoids in blood samples, food products, natural product extracts, and USP L6 commercial preparations perates with low aqueous or non aqueous mobile phases desirable in LC/MS and prep fraction recovery Usable ph range :.~7. Carotenoid analytical column YMC Carotenoid is C bonded silica based reversed-phase column. It is for carotenoid analysis and useful for separation of geometric isomers. Application (AA) Carotene and xanthophyll 6 7 8 9. Astaxanthin. Capsanthin. Lutein. Zeaxanthin. Canthaxanthin 6. β-cryptoxanthin 7. Echinenone 8. -cis β-carotene 9. -cis β-carotene. α-carotene. trans β-carotene. 9-cis β-carotene. δ-carotene. Lycopene 6 7 8 min Column :YMC Carotenoid X.6 mmi.d. Eluent :A) methanol/mtbe*/h (8//) B) methanol/mtbe*/h (6/9/) -%B (-9 min) Flow rate :. ml/min Temperature :ambient Detection : UV at nm *methyl tert-butyl ether
rdering Information -Columns- YMC-Pack Pro C8 Å µm Å µm 7 I.D. (mm) mm length. SS-WT SS-LWT SS-WT SS-WT. SS-QGC. SS-WT SS-WT SS-WT. SS-GC.6 SS-6WT SS-L6WT SS-6WT SS-6WT. SS-GC. SS-WT SS-LWT SS-WT SS-WT. SS-QGC. SS-WT SS-WT SS-WT. SS-GC.6 SS-6WT SS-L6WT SS-6WT SS-6WT SS-6WT 6. SS-6WT. SS-GC YMC-Pack Pro C Å µm Å µm 7 I.D. (mm) mm length. BSS-WT BSS-LWT BSS-WT BSS-WT. BSS-QGC. BSS-WT BSS-WT BSS-WT. BSS-GC.6 BSS-6WT BSS-L6WT BSS-6WT BSS-6WT. BSS-GC. BSS-WT BSS-LWT BSS-WT BSS-WT. BSS-QGC. BSS-WT BSS-WT BSS-WT. BSS-GC.6 BSS-6WT BSS-L6WT BSS-6WT BSS-6WT BSS-6WT 6. BSS-6WT. BSS-GC YMC-Pack C 8 Å µm Å µm Å µm Å µm YMC-Pack C Å µm Å µm Å µm 7 I.D. (mm) mm length. CS-WT CS-LWT CS-WT CS-WT. CS-QGC. CS-WT CS-WT CS-WT. CS-GC.6 CS-6WT CS-6WT. CS-GC. CS-WT CS-WT. CS-QGC.6 CS-L6WT CS-6WT CS-6WT CS-6WT 6. CS-6WT CS-6WT CS-6WT. CS-GC CS-WT CS-WT CS-CC.6 CS-6WT CS-6WT. CS-GC. CS-WT CS-WT. CS-QGC.6 CS-L6WT CS-6WT CS-6WT CS-6WT. CS-GC 6. CS-6WT CS-6WT CS-6WT CS-WT CS-WT CS-CC 7 I.D. (mm) mm length. BUS-WT BUS-LWT BUS-WT BUS-WT. BUS-QGC. BUS-WT BUS-WT BUS-WT. BUS-GC.6 BUS-6WT BUS-6WT. BUS-GC. BUS-WT BUS-WT. BUS-QGC.6 BUS-L6WT BUS-6WT BUS-6WT BUS-6WT 6. BUS-6WT BUS-6WT BUS-6WT. BUS-GC BUS-WT BUS-WT BUS-CC. BUS-WT BUS-WT. BUS-QGC.6 BUS-L6WT BUS-6WT BUS-6WT BUS-6WT 6. BUS-6WT BUS-6WT BUS-6WT. BUS-GC BUS-WT BUS-WT BUS-CC * Guard cartridge holder required, part no. XPGCH-Q for. -. mmi.d. and XPCHSPW for mmi.d. See P. for preparative columns other than those listed above.
7 rdering Information -Columns- YMC-Pack TMS Å µm Å µm 7 I.D. (mm) mm length.6 TMS-6WT TMS-6WT. TMS-GC. TMS-WT TMS-WT. TMS-QGC.6 TMS-L6WT TMS-6WT TMS-6WT TMS-6WT. TMS-GC 6. TMS-6WT TMS-6WT TMS-6WT TMS-WT TMS-WT TMS-CC YMC-Pack Ph Å µm Å µm 7 I.D. (mm) mm length. PHS-WT PHS-LWT PHS-WT PHS-WT. PHS-QGC. PHS-WT PHS-WT PHS-WT. PHS-GC.6 PHS-6WT PHS-6WT. PHS-GC. PHS-WT PHS-WT. PHS-QGC.6 PHS-L6WT PHS-6WT PHS-6WT PHS-6WT 6. PHS-6WT PHS-6WT PHS-6WT. PHS-GC PHS-WT PHS-WT PHS-CC YMC-Pack CN Å µm Å µm Å µm YMC-Pack PRTEIN-RP Particle size µm Column I.D. (mm) 7 I.D. (mm) mm length. CNS-WT CNS-LWT CNS-WT CNS-WT. CNS-QGC. CNS-WT CNS-WT CNS-WT. CNS-GC.6 CNS-6WT CNS-6WT. CNS-GC. CNS-WT CNS-WT. CNS-QGC.6 CNS-L6WT CNS-6WT CNS-6WT CNS-6WT 6. CNS-6WT CNS-6WT CNS-6WT. CNS-GC CNS-WT CNS-WT CNS-CC. CNS-WT CNS-WT. CNS-QGC.6 CNS-L6WT CNS-6WT CNS-6WT CNS-6WT 6. CNS-6WT CNS-6WT CNS-6WT. CNS-GC 7 I.D. (mm) mm length. PR99S-WT PR99S-WT. PR99S-QGC.6 PR99S-6WT PR99S-6WT. PR99S-GC PR99S-WT PR99S-QGC YMCbasic Å µm Å µm 7 I.D. (mm) mm length. BA99S-WT BA99S-LWT BA99S-WT BA99S-WT. BA99S-QGC. BA99S-WT BA99S-WT BA99S-WT. BA99S-GC.6 BA99S-6WT BA99S-6WT BA99S-6WT. BA99S-GC. BA99S-WT. BA99S-QGC. BA99S-WT. BA99S-GC.6 BA99S-6WT BA99S-6WT BA99S-6WT BA99S-6WT 6. BA99S-6WT BA99S-6WT. BA99S-GC YMC Carotenoid Particle size Column I.D. (mm) 7 I.D. (mm) mm length µm.6 CT99S-6WT CT99S-6WT -. CT99S-GC µm.6 CT99S-6WT CT99S-6WT. CT99S-GC * Guard cartridge holder required, part no. XPGCH-Q for. -. mmi.d. and XPCHSPW for mmi.d. See P., for preparative columns other than those listed above.