Reversed-Phase C18 Columns (ODS)
Transcription
Reversed-Phase C18 Columns (ODS)
YMC GENERAL CATALOG 1 Reversed-Phase C18 Columns (ODS) Types and characteristics of C18 packing materials 14 Guides for selecting C18 packing materials 15 YMC-Triart C18 16∼23 YMC-UltraHT series YMC-UltraHT series YMC-UltraHT Pro C18 YMC-UltraHT Hydrosphere C18 24,25 26 27 Pro series Features of Pro series ODS columns YMC-Pack Pro C18 Hydrosphere C18 YMC-Pack Pro C18 RS 28∼31 32∼35 36∼39 40∼43 YMC-Pack ODS series YMC-Pack YMC-Pack YMC-Pack YMC-Pack ODS-A ODS-AM ODS-AQ ODS-AL 44,45 46,47 48,49 50,51 , J sphere ODS series ODS-H80、M80、L80 52,53 YMC-Pack PolymerC18 54,55 13 第01章.indd 13 11.1.28 10:39:03 AM Types and characteristics of C18 packing materials Octadecyl-silica (ODS) is the industry standard packing material for HPLC applications. YMC Co., Ltd. offers packing materials with an impressive assortment of functional groups for liquid chromatography and a selection of ODS columns that is far ahead of all others in variety. We also offer hybrid-silica based C18 packing materials that are widely applicable to a variety of fields, from microanalysis to large-scale isolation. Types of C18 packing materials Base Products Hybrid silica ( ODS ) Reversed-Phase C18 Columns 1 YMC-Triart C18 Silica YMC-UltraHT series YMC-Pack Pro C18 Hydrosphere C18 YMC-Pack Pro C18 RS YMC-Pack ODS series J’ sphere ODS series Polymer YMC-Pack PolymerC18 Types and characteristics of C18 (ODS) packing materials Product name ODS Pore Particle Usable size size Silanol pH range (Å) (µm) C% treatment 2 YMC-Triart C18 120 3 20 Yes 5 120 2 16 Yes Hydrosphere C18 120 Pro series Pro C18 120 Hydrosphere C18 120 Pro C18 RS YMC-Pack ODS series 80 120 ODS-A 200 300 ODS-AM ODS-AQ 120 120 200 ODS-AL J'sphere ODS series ODS-H80 ODS-M80 ODS-L80 YMC-Pack PolymerC18 120 80 80 80 — 2 3 5 3 5 3 5 3 5 10 5 10 5 10 3 5 3 5 10 5 10 5 4 4 4 6 10 Pages ・Versatile hybrid silica based ODS column 1.0 〜 12.0 16 〜 23 ・Great chemical durability ・Excellent peak shape in every conditions UltraHT series Pro C18 Characteristics 2.0 〜 8.0 12 ・2 µm-ODS column for Ultrafast LC method ・Superior separation of basic compounds 26 ・Superior separation of hydrophilic compounds ・Reduced analysis time while maintaining resolution 27 Yes ・Processed with YMC Co., Ltd.’s advanced endcapping technology 32 〜 35 ・Superior separation of basic compounds 12 Yes 2.0 〜 8.0 ・Superior separation of hydrophilic compounds 36 〜 39 ・Can be used with 100% water mobile phase 22 Yes ・Highly durable ODS 1.0 〜 10.0 ・Superior separation of basic compounds 40 〜 43 and hydrophobic compounds 16 17 12 Yes 7 17 14 Yes Yes 10 17 22 14 9 — No Yes Yes Yes — — 2.0 〜 7.5 ・Currently in use worldwide 44,45 ・ODS with wide pore size ・For separation of peptides and proteins 44,45 ・Outstanding lot-to-lot reproducibility 46,47 ・Superior separation of hydrophilic compounds 48,49 ・For separation utilizing residual silanol 1.0 〜 9.0 ・High carbon ODS ・Medium carbon ODS 2.0 〜 7.5 ・Low carbon ODS 2.0 〜 13.0 ・Polymer-based ODS 50,51 52,53 54,55 14 第01章.indd 14 11.1.28 10:39:04 AM Guides for selecting C18 packing materials YMC Co., Ltd. offers twelve types of ODS columns, each with unique separation characteristics. The proper selection of packing material is therefore a key factor in the establishment of efficient separation conditions. Selection of packing materials without sufficient consideration of the separation characteristics may result in an expense of much time and effort in separating a target material. Low molecular weight compounds High molecular weight compounds Chemical structures differ slightly among solutes YMC-Pack Pro C18 RS J’sphere ODS-H80 Hydrophobicity differs greatly among solutes YMC-Pack ODS-A (Wide-Pore) J’sphere ODS-L80 Difficult to separate using standard ODS Hydrosphere C18 YMC-Pack Pro C18 RS YMC-Pack ODS-AL J’sphere ODS-H80 J’sphere ODS-L80 YMC-Pack PolymerC18 Basic compounds YMC-Triart C18 YMC-Pack Pro C18 Hydrosphere C18 YMC-Pack Pro C18 RS YMC-Pack ODS-AM YMC-Pack PolymerC18 Coordination compounds YMC-Triart C18 YMC-Pack Pro C18 Hydrosphere C18 YMC-Pack Pro C18 RS YMC-Pack ODS-AM YMC-Pack PolymerC18 Sugars and glycosides Hydrosphere C18 YMC-Pack ODS-AQ J’sphere ODS-L80 High molecular weight peptides and oligonucleic acids YMC-Pack ODS-A (Wide-Pore) Alkaline mobile phase ( ODS ) Reversed-Phase C18 Columns General organic compounds YMC-Triart C18 YMC-Pack Pro C18 YMC-Pack ODS-A J’sphere ODS-M80 1 YMC-Triart C18 YMC-Pack Pro C18 RS J’sphere ODS-H80 YMC-Pack PolymerC18 15 第01章.indd 15 11.1.28 10:39:05 AM Versatile Hybrid Silica Based HPLC Column YMC-Triart C18 YMC-Triart C18 is a hybrid silica based ODS column emphasizing durability, low operating pressure, and excellent performance. YMC-Triart C18 has been developed and is manufactured using state-of-the-art technologies that define materials, granulation and surface modification. Rugged, long lasting, hybrid particle technology Integration of triple artistic technologies Benefit ● Great chemical durability ● Lower back pressure ● Excellent resolution ▼ U sable over a wide pH range makes the column excellent for method development. Long column lifetime over wide pH range. Low cost per analysis. ▼ ● Hybrid silica base A bility to use on both older and newer instrumentation. ▼ Features R ugged, first choice option for analysis of acids, bases and neutral compounds. ▼ ( ODS ) Reversed-Phase C18 Columns 1 Specifications Base :organic/inorganic hybrid silica Carbon % Stationary :C18 (as USP-L1) Bonding :polymeric type Particle :2 µm, 3 µm, 5 µm Endcapping :multi-stage endcapping Pore :120 Å pH range :pH 1 〜 12 :20% YMC-Triart C18 -Integration of triple artistic technologies- <1>Versatile hybrid material YMC-Triart C18 is a multi-layer hybrid particle consisting of two types of layers: a silica based layer (inorganic) and a hybrid polymer based layer (organic). This layered structure contributes to Triart's combined physical durability and chromatographic performance. inorganic-silane layer Particle synthesis by Microreaction Versatile hybrid silica material Multi-stage endcapping Image structure for hybrid-silica gel organic-silane layer 16 第01章.indd 16 11.1.28 10:39:08 AM YMC-Triart C18 -Integration of triple artistic technologies- <2>New granulation process by microreactor technology Homogeneous and uniform particle Narrow pore distribution Triart C18 4500 4000 Hybrid particle for Triart 3000 Silica particle for conventional product 2500 2000 Hybrid particle for competitor's product 1500 1000 500 0 Use of microreactor technology produces homogeneous and uniform particles that results in low operating pressure and very reproducible surface modification making for the ultimate in column-to-column and lot-to-lot reproducibility. 0 50 100 150 200 Pore size (Å) 250 300 350 YMC-Triart C18 has a very narrow pore size distribution that enables narrow and reproducible peak shapes for analytes. <3>Surface modification technology Multi-stage endcapping ( ODS ) Reversed-Phase C18 Columns d Vp/d logD 3500 1 Endcapping methods employed for production of conventional products typically employ a single endcapping compound in one chemical step to "neutralize" highly reactive silanols. Unfortunately, highly reactive silanols are easily hydrolyzed and contribute to poor durability. Additionally, low reactive silanols endcap stubbornly, producing a surface modification that is often non-reproducible, and leading to peak tailing. YMC-Triart C18 solves this problem by using an innovative, newly developed "measured multi-stage/multicompound surface modification" technology which results in the thorough endcapping of all silanols that are accessible to analytes. Reaction image for "multi-stage endcapping" Evaluation of silanol activity mAU Triart C18 60 1 Before EC STEP 2 2 STEP 1 40 3 20 0 0.0 Silanol group 60 40 3 10.0 15.0 min mAU Endcapping (EC) group STEP Q090928C 5.0 Endcapping by traditional method Peak tailing for basic compounds 3 1 20 2 0 0.0 Q090928E 5.0 10.0 15.0 min Column :5 µm, 150×3.0 mmI.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)/acetonitrile (65/35) Flow rate :0.425 mL/min Temperature:40 ℃ Detection :UV at 235 nm Sample :1. Chlorpheniramine 2. Dextromethorphan 3. Propyl p-hydroxybenzoate 17 第01章.indd 17 11.1.28 10:39:14 AM Versatile Hybrid Silica Based HPLC Column YMC-Triart C18 Quality Control -Excellent reproducibility- Packing Material YMC-Triart C18 exhibits excellent lot-to-lot reproducibility for all types of compounds including difficult to chromatograph molecules that exhibit basic and coordination compound behavior. Basic compounds 2 1 Lot. E Lot. E Lot. D Lot. D Lot. C Lot. C Lot. B Lot. B Lot. A Lot. A 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 0.0 min 2. Dextromethorphan 12.0 14.0 min Column :5 µm, 150×3.0 mmI.D. Eluent :acetonitrile/0.1% H3PO4 (40/60) Flow rate :0.425 mL/min Temperature:40 ℃ Detection :UV at 254 nm 15000 2.0 10000 1.5 Tailing factor 5000 1.0 10 20 30 40 50 0.5 Low back-pressure -Free from pressure problem- Column pressure and solvents Triart C18, MeOH/W Triart C18, ACN/W Pro C18, MeOH/W 18 16 14 Pro C18, ACN/W 12 10 8 6 4 2 20 30 40 50 60 70 80 90 Standard ODS -Best balance hydrophobicity- Comparison of separation selectivity 20 10 Column :5 µm, 150×4.6 mmI.D. Eluent :acetonitrile/water (60/40) Flow rate :1.0 mL/min Temperature:ambient Sample :butyl benzoate 100 Organic solvent rate (%) Column :5 µm, 150×4.6 mmI.D. Eluent :acetonitrile/water or methanol/water Flow rate :1.0 mL/min Temperature:25 ℃ YMC-Triart C18 is designed for use in all conditions. Even with higher viscosity methanol, YMC-Triart C18 can operate with lower pressure (typically 30% less than conventional columns). Hydrogen-bonding capacity α (caffeine/benzene) Theoretical plate number 10.0 2.5 Column number Column pressure (MPa) 8.0 Theoretical plate number 0 0 6.0 Rigid control of theoretical plate number (N) and tailing factor (Tf) are exhibited for each YMC-Triart C18 packed column. 20000 0 4.0 1. Hinokitiol 2. Methyl benzoate Column :5 µm, 150×3.0 mmI.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)/acetonitrile (65/35) Flow rate :0.425 mL/min Temperature:40 ℃ Detection :UV at 235 nm Packed Column 2.0 J091201X J091201Y 1. Chlorpheniramine 3. Propyl p-hydroxybenzoate 2 1 3 Tailing factor ( ODS ) Reversed-Phase C18 Columns 1 Coordination compounds 0.180 0.160 Develosil C30-UG Hydrosphere C18 0.140 Atlantis T3 CAPCELL PAK C18 AQ 0.120 Standard ODS CAPCELL PAK C18 MGIII Gemini C18 The most versatile columns as first-choice. Pro C18 Unison UK-C18 Inertsil ODS-4 Hypersil GOLD CAPCELL PAK C18 MGII Gemini NX-C18 SunFire Inertsil ODS-3 XBridge C18 XTerra MS C18 Mightysil RP-18 C18 Eternity-C18 L-column ODS 0.100 Triart C18 0.080 TSKgel ODS-100S Cadenza CD-C18 L-column2 ODS Develosil ODS-SR Pro C18 RS 0.060 Eluent : MeOH/W (80/20) [amylbenzene], MeOH/W (30/70) [caffeine, benzene] 0.040 2.00 4.00 6.00 8.00 10.00 Hydrophobicity k’ (amylbenzene) 12.00 14.00 (S090805A) Conventional hybrid silica based ODS columns tend to be less hydrophobic than silica based columns. YMC-Triart C18 has a favorable carbon balance and may be used as a "versatile firstchoice" method development column. 18 第01章.indd 18 11.1.28 10:39:17 AM Great durability -Wide usable pH range and long lifetime- Durability in high pH Phosphate buffer (pH 11.5, 40 ℃) Triethylamine (pH 11.5, 40 ℃) Rate of theoretical plate number 120% YMC-Triart C18 100% 80% 60% 40% Hybrid-silica based C18 Silica based C18 20% 0% 0 10 20 30 40 80% 60% 40% Silica based C18 20% 0% 50 YMC-Triart C18 100% 0 50 100 Time (hr) 150 200 250 ( ODS ) Reversed-Phase C18 Columns Rate of theoretical plate number 120% 300 Time (hr) Column :5 µm, 150×4.6 mmI.D. Eluent :50 mM K2HPO4-K3PO4 (pH 11.5)/methanol (90/10) Flow rate :1.0 mL/min Temperature:40 ℃ Sample :benzyl alcohol Column :5 µm, 150×4.6 mmI.D. Eluent :50 mM triethylamine (pH 11.5)/methanol (90/10) Flow rate :1.0 mL/min Temperature:40 ℃ Sample :benzyl alcohol Durability in high temperature pH 6.9, 70 ℃ pH 1, 70 ℃ Rate of theoretical plate number 120% 120% YMC-Triart C18 100% 100% Rate of retention time YMC-Triart C18 80% Silica based C18 alkaline-resistant type 60% Silica based C18 40% Silica based C18 80% 40% 20% 0% 0% 100 200 300 400 500 600 700 Hybrid-silica based C18 60% 20% 0 1 The columns are kept in acetonitrile/water/TFA (10/90/1), 70 °C, and tested for performance every 20 hours. 0 20 40 60 80 100 120 Time (hr) Time (hr) Column :5 µm, 50×2.0 mmI.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)/acetonitrile (90/10) Flow rate :0.2 mL/min Temperature:70 ℃ Sample :phenol Test conditions Column :5 µm, 50×2.0 mmI.D. Eluent :acetonitrile/water (60/40) Flow rate :0.2 mL/min Temperature:37 ℃ Sample :butyl benzoate YMC-Triart C18 shows great chemical stability provided by newly developed hybrid silica. Even in high pH or high temperature conditions, the lifetime of YMC-Triart C18 is more than 10 times greater than conventional ODS columns. Reduced column bleed -Operate at high sensitivity- Bleed test by LC/MS Scan ES+ TIC 1.2E+09 − Triart C18 − No column 8.0E+08 Column Eluent 4.0E+08 0.0E+00 0 2 4 6 8 10 12 min :5 µm, 50×2.0 mmI.D. :A) water/formic acid (100/0.1) B) acetonitrile/formic acid (100/0.1) 5% B (0-1 min), 5-100% B (1-5 min), 100% B (5-10 min), 100-5% B (10-10.1 min), 5% B (10.1-12.5 min) Flow rate :0.4 mL/min Temperature:40 ℃ Detection :ESI positive, TIC (Mass Range: 50-1000) Bleeding of stationary phase can often contribute to high background signals when employing MS and other hyphenated detection techniques. In the example cited above, use of a 50×2.0 mmI.D. YMC-Triart C18 column shows no contribution to the signal observed by the MS positive total ion current (TIC) trace. Such performance allows for low limits of detection (high S/N ratio) for MS and other detection techniques. The inherent hybrid particle strength is also superior to conventional silica materials for detection techniques that are sensitive to particle degradation. 19 第01章.indd 19 11.1.28 10:39:21 AM Versatile Hybrid Silica Based HPLC Column YMC-Triart C18 Excellent resolution -Without adsorption and tailing- Analysis of basic compounds Ingredients in a cough/cold medication YMC-Triart C18 2 1 5 µm, 150×3.0 mmI.D. 1. Cl CH3 O 2. CH3 N H3C 3 N ( ODS ) Reversed-Phase C18 Columns 1 N H Q090928C CH3 Chlorpheniramine 1 2 Silica based C18 Brand D5 3 5 µm, 150×4.6 mmI.D. 3. O H3C O Q090821C 3 2 Propyl p-hydroxybenzoate Silica based C18 Brand N1 1 Column Eluent Flow rate :150×3.0 mmI.D. or 150×4.6 mmI.D. :20 mM KH2PO4-K2HPO4 (pH 6.9)/acetonitrile (65/35) :0.425 mL/min for 3.0 mmI.D. 1.0 mL/min for 4.6 mmI.D. Temperature:40 ℃ Detection :UV at 235 nm 3 µm, 150×3.0 mmI.D. J091111K 1 2 Hybrid-silica based C18 Brand I8 3 Dextromethorphan OH 5 µm, 150×4.6 mmI.D. Q090123F 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min Analysis of coordination compounds YMC-Triart C18's innovative surface modification technology results in excellent peak shapes even for the basic compounds that often exhibit tailing shapes on conventional silica and hybrid silica based ODS columns. Hinokitiol CH3 1. CH3 2. O O CH3 2 1 YMC-Triart C18 5 µm, 150×3.0 mmI.D. OH O Q090925G Silica based C18 Brand N1 2 1 3 µm, 150×3.0 mmI.D. J091110B 2 Hybrid-silica based C18 Brand I8 5 µm, 150×4.6 mmI.D. 1 0.0 2.0 4.0 Q090120A 6.0 8.0 10.0 12.0 14.0 min Analysis of acidic compounds YMC-Triart C18 5 µm, 150×3.0 mmI.D. 2 Q090605C 1 5 µm, 150×4.6 mmI.D. 3 Q090717A 1.5 2.0 :150×3.0 mmI.D. or 150×4.6 mmI.D. :acetonitrile/0.1% H3PO4 (40/60) :0.425 mL/min for 3.0 mmI.D. 1.0 mL/min for 4.6 mmI.D. Temperature:40 ℃ Detection :UV at 254 nm YMC-Triart C18 has an extremely low level of metal impurities, much lower than conventional products. YMC-Triart C18 is able to provide excellent peak shape for coordination compounds. 1. Formic acid 2. Acetic acid 3. Propionic acid :150×3.0 mmI.D. or 150×4.6 mmI.D. :acetonitrile/0.1% H3PO4 (5/95) :0.425 mL/min for 3.0 mmI.D. 1.0 mL/min for 4.6 mmI.D. Temperature:37 ℃ Detection :UV at 210 nm Hybrid-silica based C18 Brand G4 2 1.0 Column Eluent Flow rate Column Eluent Flow rate 3 0.5 Methyl benzoate Organic acid 1 0.0 Hinokitiol 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 min YMC-Triart C18 is synthesized utilizing methodology borrowed from microreactor technology. This synthesis technique allows for a reduction of impurities that contribute to peak tailing during the analysis of some types of acidic compounds. 20 第01章.indd 20 11.1.28 10:39:24 AM Excellent versatility -Buffer independency- Comparison of clemastine analysis with conventional columns 10 mM phosphate buffer (pH 6.7)/organic solvent methanol 10 mM CH3COONH4/organic solvent methanol acetonitrile acetonitrile YMC-Triart C18 50×2.0 mmI.D. Tf 0.97 Tf 1.11 Tf 1.21 50×2.1 mmI.D. Tf 1.56 F090727H07 Silica based C18 Brand D5 Silica based C18 Brand I6 Tf 1.03 Hybrid-silica based C18 Brand I8 Tf 1.14 50×2.1 mmI.D. 10.0 H3C N 11.0 12.0 Tf 1.78 F090731G07 F090731F07 min 8.0 9.0 10.0 min 6.0 7.0 8.0 min 1 Column Eluent :5 µm, 50×2.0 mmI.D. or 50×2.1 mmI.D. :A) 10 mM KH2PO4-K2HPO4 (pH 6.7) or 10 mM CH3COONH4 (pH 6.7) B) methanol or acetonitrile 5-90% B (0-10 min), 90% B (10-15 min) Flow rate :0.2 mL/min Temperature:25 ℃ Detection :UV at 230 nm CH3 O 8.0 F090722G07 Tf 1.22 F090731I07 7.0 Tf 1.88 F090722F07 Tf 1.88 6.0 F090714G07 Tf 1.13 F090722I07 min Tf 2.21 F090714F07 Tf 3.87 F090731H07 50×2.1 mmI.D. F090803G07 Tf 1.05 F090717I07 F090722H07 Tf 1.43 F090727F07 Tf 2.83 F090717H07 F090817G07 Tf 1.13 F090727I07 Tf 0.93 50×2.0 mmI.D. Tf 1.16 F090818F07 ( ODS ) Reversed-Phase C18 Columns Silica based C18 Brand C1 Tf 1.03 F090818I07 F090818H07 Cl Clemastine Clemastine is a well known basic compound which can easily tail on conventional ODS columns. YMC-Triart C18 can separate clemastine sharply using many buffer/solvent combinations. Improvement of loadability -Minimizing strong solvent/sample loading effects- Influence on peak shapes by injection volume 1 2 1 Hybrid-silica based C18 2 1 2 solvent for sample injection volume acetonitrile 1.0 µL 1.5 µL 2.0 µL 3.0 µL YMC-Triart C18 Column Eluent :5 µm, 50×2.0 mmI.D. or 2.1 mmI.D. :A) water/formic acid (100/0.1) B) acetonitrile/formic acid (100/0.1) 5% B(0-0.5 min) , 5-100% B (0.5-2.5 min) Flow rate :0.4 mL/min Temperature:40 ℃ Detection :UV at 275 nm Silica based C18 1. Peak anomaly at 1.5 µL 2. OCH2CHCH2NHCH(CH3)2 OH H3CO H3CO CN CH3 C (CH2)3NCH2CH2 OCH3 OCH3 CH(CH3)2 1.5 2.0 2.5 min Propranolol (50 µg/mL) Verapamil (50 µg/mL) YMC-Triart C18 can tolerate larger injection volumes of samples containing strong solvents (e.g., acetonitrile) while allowing for better peak shape than traditional columns. This can be important for difficult to solubilize samples that need higher concentrations of organic solvent for solubilization prior to analysis. 21 第01章.indd 21 11.1.28 10:39:27 AM Versatile Hybrid Silica Based HPLC Column YMC-Triart C18 Fast analysis of paraquat and diquat YMC-Triart C18 5 µm, 150×2.0 mmI.D. YMC-Triart C18 5 µm, 150×2.0 mmI.D. mAU 10%B, 0.2 mL/min, 5.7 MPa 1 25 20 ( ODS ) Reversed-Phase C18 Columns 1 mAU 2 20 Eluent optimization Insufficient separation 15 10 5 5 0 0 R091126J 2 4 6 8 Baseline separation, But long analysis time 15 10 0 5%B, 0.2 mL/min, 5.6 MPa 25 10 12 14 min 1 2 R091127C 0 5 10 15 20 25 min Downsizing 1. YMC-Triart C18 3 µm, 50×2.0 mmI.D. 2. N+ N+ 1 mAU ·2Br− 2 N+ CH3 ·2Cl− H3C N+ 25 20 Paraquat Diquat 5%B, 0.4 mL/min, 12.2 MPa 80% reduction in run time 15 10 Eluent :A) water/HFBA* (100/0.1) B) acetonitrile/HFBA* (100/0.1) Temperature:37 ℃ Detection :UV at 290 nm Injection :1 µL (0.1 mg/mL) *heptafluorobutyric acid 5 0 R091201B 0 1 2 3 Application 4 5 min (Q091110E) Anthocyanin 1. Delphinidin-3-O-galactoside 2. Delphinidin-3-O-glucoside 3. Cyanidin-3-O-galactoside 4. Delphinidin-3-O-arabinoside 5. Cyanidin-3-O-glucoside 6. Petunidin-3-O-galactoside 7. Cyanidin-3-O-arabinoside 8. Petunidin-3-O-glucoside 9. Peonidin-3-O-galactoside 10. Petunidin-3-O-arabinoside 11. Delphinidin 12. Peonidin-3-O-glucoside 13. Malvidin-3-O-galactoside 14. Peonidin-3-O-arabinoside 15. Malvidin-3-O-glucoside 16. Malvidin-3-O-arabinoside 17. Cyanidin 18. Petunidin 19. Peonidin 20. Malvidin 1 mAU 2 100 80 4 5 60 3 8 7 40 15 6 16 17 12 20 10 11 13 9 18 20 19 14 0 0 5 10 15 20 25 30 35 40 min Column :YMC-Triart C18 (5 µm, 120 Å) 250×4.6 mmI.D. Eluent :A) water/formic acid (90/10) B) acetonitrile/methanol/water/formic acid (22.5/22.5/40/10) 20-28% B (0-30 min), 28-70% B (30-40 min), 100% B (40-45 min) Flow rate :1.0 mL/min Temperature:25 ℃ Detection :UV/VIS at 535 nm Sample :commercial bilberry powder (1.25 mg/mL) 22 第01章.indd 22 11.1.28 10:39:38 AM Application (R091009C) Peptide mAU 1. Oxytocin 2. Met-Enkephalin 3. Leu-Enkephalin 4. Neurotensin 5. γ-Endorphin 6. β-Endorphin 3 100 6 1 75 4 2 50 Column 5 25 0 5 10 15 25 min 20 :YMC-Triart C18 (5 µm, 120 Å) 150×2.0 mmI.D. Eluent :A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 20-45% B (0-25 min) Flow rate :0.2 mL/min Temperature:37 ℃ Detection :UV at 220 nm Application (S100121A) LC/MS analysis for Triazolam and Etizolam Intens . x104 6 High resolution extracted chromatograms 1. + for[M+H] H3C N N N 5 Cl 4 2. N N Mixture 100 ppb 2 H3C 1 S H3C N N N Cl Cl 3 Triazolam (MW 343.22) 2 Etizolam (MW 342.85) Column 1 0 0 2 4 6 8 10 12 14 Time [min] 100ng_mL_RB4_01_2291.d: EIC 343.051±0.01 +All MS 100ng_mL_RB4_01_2291.d: EIC 343.078±0.01 +All MS Courtesy of J.Watanabe, Bruker Daltonics K. K. :YMC-Triart C18 (5 µm, 120 Å) 50×2.0 mmI.D. Eluent :A) 10 mM HCOOH, B) acetonitrile 25-50% B (0-10 min), 25% B (10-15 min) Flow rate :0.2 mL/min Temperature:40 ℃ Detection :ESI, positive mode (Bruker Daltonics micrOTOF) Injection :5 µL ( ODS ) Reversed-Phase C18 Columns 0 (MW 1,007) (MW 574) (MW 556) (MW 1,673) (MW 1,859) (MW 3,465) 1 Ordering information Analytical columns Particle size (µm) S-3 S-5 Semi-preparative columns Pore size Column size (Å) inner diameter×length(mm) 120 120 Please inquire for 2 µm columns. 2.0× 20 2.0× 30 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6× 35 4.6× 50 4.6× 75 4.6×100 4.6×150 2.0× 20 2.0× 30 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0× 75 3.0×100 3.0×150 4.6× 50 4.6× 75 4.6×100 4.6×150 4.6×250 Product number TA12S03-0202WT TA12S03-0302WT TA12S03-0502WT TA12S03-L502WT TA12S03-1002WT TA12S03-1502WT TA12S03-0503WT TA12S03-1003WT TA12S03-1503WT TA12S03-H546WT TA12S03-0546WT TA12S03-L546WT TA12S03-1046WT TA12S03-1546WT TA12S05-0202WT TA12S05-0302WT TA12S05-0502WT TA12S05-L502WT TA12S05-1002WT TA12S05-1502WT TA12S05-0503WT TA12S05-L503WT TA12S05-1003WT TA12S05-1503WT TA12S05-0546WT TA12S05-L546WT TA12S05-1046WT TA12S05-1546WT TA12S05-2546WT Particle size (µm) S-5 Pore size Column size (Å) inner diameter×length(mm) 120 10×150 10×250 20×100 20×150 20×250 Product number TA12S05-1510WT TA12S05-2510WT TA12S05-1020WT TA12S05-1520WT TA12S05-2520WT Semi-preparative columns YMC-Actus Triart C18 TA12S05-0520WX 20× 50 S-5 120 20×100 30× 50 30× 75 30×100 TA12S05-1020WX TA12S05-0530WX TA12S05-L530WX TA12S05-1030WX See P138-141 for details of YMC-Actus series. Guard cartridge columns (with inner diameter 2.0 mm: 2/pack, 4.0 mm: 3/pack) TA12S03-0102CC 2.0×10 S-3 120 S-5 120 4.0×23 2.0×10 4.0×23 TA12S03-G304CC TA12S05-0102CC TA12S05-G304CC A cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) Guard columns S-5 120 10×30 20×50 XPCHSMW XPCHW TA12S05-0310WTG TA12S05-0520WTG 23 第01章.indd 23 11.1.28 10:39:40 AM Ultra-Fast LC column YMC-UltraHT ( ODS ) Reversed-Phase C18 Columns 1 YMC-UltraHT series is a C18 bonded-phase based on standard ODS, and Hydrosphere C18 for hydrophilic highly efficient 2 µm spherical silica particles. YMC- compounds. Both of them show the same selectivity UltraHT series columns are specifically designed for and superior peak shapes as 3 µm and 5 µm Pro series, high-speed and high throughput analysis in various fields allowing for an easy method transfer from conventional such as pharmaceuticals, foods, and environmentals. HPLC to ultra-fast HPLC without changing elution YMC-UltraHT series has two products. Pro C18 as our conditions. 2 µm ODS column ideal for Ultra-Fast LC Features ● Superior column performance at higher flow rate and higher pressure ● Reduced analysis time while maintaining excellent resolution ● Excellent resolution with back pressure less than that of sub-2 µm ● Applicable to both conventional HPLC and specific ultra pressure LC system ● Same selectivity and superior peak shapes as 3 µm and 5 µm Pro series ●S imple method transfer from conventional HPLC on Pro series; no need to change elution conditions Characteristics of 2 µm packing material Van Deemter curves Column pressure 0.020 80 Optimum flow rate for 5 µm Optimum flow rate for 2 µm (0.3-0.8 mL/min for 2 mmI.D.) 0.015 YMC 3 µm 0.010 Brand L1 1.8 µm YMC 2 µm 0.005 Optimum flow rate for 2 µm 70 YMC 5 µm Column pressure (MPa) HETP (mm/plates) (0.2 mL/min for 2 mmI.D.) Brand I7 1.7 µm Brand I7 1.7 µm (0.3-0.8 mL/min for 2 mmI.D.) Brand L1 1.8 µm 60 50 40 YMC 2 µm applicable pressure range for conventional LC 30 YMC 3 µm 20 YMC 5 µm 10 0.000 0 1 2 3 4 5 6 7 8 9 10 0 11 Interstitial linear velocity (mm/sec) 0 1 2 3 4 5 6 7 8 9 10 11 Interstitial linear velocity (mm/sec) YMC-UltraHT series shows nearly the same efficiency as sub-2 µm columns for ultra fast LC system and reduces the column pressure significantly. Column : 50×2.0 or 2.1 mmI.D. Eluent : acetonitrile/water (60/40) Temperature: 25 ℃ Sample : Butyl benzoate YMC-UltraHT series Pro C18 Particle Size (µm) Pore Size (Å) Hydrosphere C18 2 2 120 120 Specific Surface area (m2/g) 330 330 Carbon Content 16% 12% Monomeric Monomeric Bonding Type Available pH Range Characteristics 2-8 2-8 Standard type For polar analytes 24 第01章.indd 24 11.1.28 10:39:43 AM Ultra Fast analysis for flavonoids conventional LC method:YMC-Pack C18 150×4.6 mmI.D. 5 µm tR(7) 24.3 min N(7) 77,000 /m 1.0 mL/min 5 µL injection 8.5 MPa Ultra-fast LC method:YMC-UltraHT C18 50×2.0 mmI.D. 2 µm Flavonoids 90% Decrease in analysis time 90% Increase in efficiency 1. Myricetin 2. Quercetin 3. Apigenin 4. Kaempferol 5. Baicalein 6. Chrysin 7. Acacetin tR(7) 2.7 min N(7) 144,000 /m 0.6 mL/min 1 µL injection 30 MPa Eluent : acetonitrile/water/formic acid (35/65/0.1) Temperature: 40 ℃ Detection : UV at 260 nm ( ODS ) Reversed-Phase C18 Columns Triplear line city Velo 1 This example shows the method transfer from conventional HPLC method to Ultra-fast LC method using a YMC-UltraHT column. Since the column efficiency is not decreased even when the flow rate is 3 times as fast as the conventional condition, it can be shorten analysis time by 90% without losing the separation characteristics. Applicable for conventional LC system 50×3 mmI.D. Conventional semi-micro LC system(2.5 µL, cell length 5 mm) mAU 1 80 Flow rate : 0.8 mL/min Injection : 2 µL 2 3 60 4 40 20 0 50×2 mmI.D. Conventional semi-micro LC system(2.5 µL, cell length 5 mm) 80 N(1) = 6300 N(2) = 6300 N(3) = 7500 N(4) = 7800 Rs(1, 2) = 2.21 2 mAU 1 3 2 100 4 50 0 2 3 min N(1) = 3500 N(2) = 3800 N(3) = 6100 N(4) = 6800 Rs(1, 2) = 1.61 Flow rate : 0.36 mL/min Injection : 1 µL mAU 150 3 min YMC-UltraHT series can be used for fast analysis with conventional LC system, as its pressure is lower than commercial sub-2 µm columns. Considering the extra column band broadening, the column of 3.0 mml.D. is more applicable for conventional LC system than the column of 2.0 mml.D.. 3 12 100 50 0 2 1 Conventional LC system(8 µL, cell length 10 mm) (N070606E) 1 N(1) = 3700 N(2) = 4100 N(3) = 6100 N(4) = 6900 Rs(1, 2) = 1.61 (N070608E) 0 Flow rate : 0.8 mL/min Injection : 2 µL 0 4 20 3 min Conventional LC system(8 µL, cell length 10 mm) 3 2 40 0 1 150 1 60 (N070606B) 0 Flow rate : 0.36 mL/min Injection : 1 µL mAU N(1) = 1400 N(2) = 1500 N(3) = 3600 4 N(4) = 4500 Rs(1, 2) = 0.90 (N070608B) 0 1 2 3 min Cephalosporin antibiotics 1. Cephalexin 2. Cefaclor 3. Cephaloglycin 4. Cephaloridine Column :YMC-UltraHT Pro C18 Eluent :acetonitrile/20 mM KH2PO4 (10/90) Temperature:37 ℃ Detection :UV at 260 nm 25 第01章.indd 25 11.1.28 10:39:45 AM Analytical columns YMC-UltraHT Pro C18 ● Superior column performance at higher flow rate and higher pressure ● Excellent resolution with back pressure less than 60% of commercial sub-2 µm ● Same selectivity and superior peak shapes as 3 µm and 5 µm Pro C18 ■Pore size:120 Å ■Carbon content:16% ■Usable pH range:2.0 〜 8.0 ● Simple method transfer from conventional HPLC First choice ODS column for Ultra-fast LC YMC-UltraHT Pro C18 is a C18 bonded-phase based on highly efficient 2 µm spherical silica particles. YMCUltraHT Pro C18 is designed for high-speed and high throughput analysis, advanced endcapping technology ( ODS ) Reversed-Phase C18 Columns 1 makes YMC-UltraHT Pro C18 highly appropriate for analysis of basic compounds. The physical properties and separation characteristics are strictly controlled, so it is highly suitable as the first choice column for ultra fast LC. Excellent peak shapes mV 60 YMC-UltraHT C18 50×2.0 mmI.D 2 µm 1 2 3 40 N(5) 106,000/m Tf(5) 1.07 30 MPa 4 5 20 mV 60 1 sub-2 µm ODS 2 4 20 0 N(5) 89,000/m Tf(5) 3.99 49 MPa 3 40 mV 60 40 0.4 0.8 1.2 1.6 2.0 [min] 1 2 4 N(5) 78,000/m Tf(5) 2.35 37 MPa 1.2 1.6 3 20 5 5 0 0 0.0 sub-2 µm ODS 0.0 0.4 0.8 1.2 1.6 Alkaloids 1. Scopolamine 2. Atropine 3. Cinchonine 4. Quinine 5. Dihydroquinine YMC-UltraHT Pro C18 exhibits greater peak shapes of alkaloids, which are highly basic compounds compared to commercial silica based sub-2 µm columns. 2.0 [min] 0.0 0.4 0.8 2.0 [min] Column : 50×2.0 or 2.1 mmI.D. Eluent :2 0 mM CH3COOH-CH3COONH4 (pH 4.9)/ acetonitrile (80/20) Flow rate : 0.6 mL/min Temperature: 40 ℃ Detection : UV at 220 nm Peptide mapping mV YMC-Pack C18 150×2.0 mmI.D. 5 µm * 0%B (0-5 min) 0-100%B (5-40 min) 100%B (40-45 min) 80 40 0 (06082204.D) 0 15 same flow rate, column length 1/3x, gradient time 1/3x mV * YMC-UltraHT C18 50×2.0 mmI.D. 2 µm 200 45 30 100 min 0%B (0-1.65 min) 0-100%B (1.65-13.35 min) 100%B (13.35-15 min) *undigested Cytochrome c 0 (N060811I) 5 0 10 15 min Analysis of a tryptic digest of cytochrome C is shown below. A 50mm length UltraHT Pro C18 column of 2 µm particle can be shorten the analysis time by one-thirds compared to a 150 mm column packed with 5 µm Pro C18. Eluent :A) acetonitrile/water/TFA (10/90/0.1) B) acetonitrile/water/TFA (35/65/0.1) Flow rate :0.2 mL/min Temperature:37 ℃ Detection :UV at 220 nm Sample :tryptic degest of Cytochrome c Ordering information Analytical columns Particle size (µm) S-2 Pore size Column size (Å) inner diameter×length(mm) 120 2.0× 30 2.0× 50 2.0× 75 2.0×100 3.0× 50 3.0× 75 3.0×100 Product number AS12S02-0302WT AS12S02-0502WT AS12S02-L502WT AS12S02-1002WT AS12S02-0503WT AS12S02-L503WT AS12S02-1003WT 26 第01章.indd 26 11.1.28 10:40:11 AM Analytical columns YMC-UltraHT Hydrosphere C18 ● Can be used with 100% aqueous as mobile phase ● Superior column performance at higher flow rate and higher pressure ● Reduced analysis time while maintaining excellent resolution ■Pore size:120 Å ■Carbon content:12% ■Usable pH range:2.0 〜 8.0 The best column for hydrophilic compounds YMC-UltraHT Hydrosphere C18 is a C18 bonded phase It is designed to maintain hydrohilicity on the packing based on highly efficient 2 µm spherical silica particles. surface for superior separation of hydrophilic compounds. Hydrosphere C18 50×4.6 mmI.D. 3 µm Rs(10,11)=2.38 3 1 9 2 4 11 6 5 12 10 7 8 Examples illustrated shows the optimization process of isoflavones separation by 2 µm YMC-UltraHT Hydrosphere C18. It has the same selectivity as 3 µm Hydrosphere C18, so there is no need to change the condition at downsizing. Maintaining resolution, the analysis time can be reduced by using 2 µm packing material. 12.5-30% B (0-10 min) 1.5 mL/min 3 µL injection (N060509E) 0 2 4 6 10 min 8 粒子サイズ、 カラム内径をダウンサイジング downsizing the particle size and column inner diameter ( ODS ) Reversed-Phase C18 Columns Ultrafast analyses of soy isoflavones 1 YMC-UltraHT Hydrosphere C18 2μm 50×2.0 mmI.D. YMC-UltraHT Hydrosphere C18 50×2.0 mmI.D. 2 µm Rs(10,11)=2.91 Rs(10,11)=2.91 Rs(7,8)=2.46 3 3 1 1 2 2 9 9 5 4 5 4 6 6 12 12 10 10 11 11 7 7 8 8 12.5-30% B (0-10 min) 0.3 mL/min 12.5-30% ACN (0-10 min) 0.6 µL injection 0.3 mL/min MPa (N070130I) (N070130I) 0 0 2 2 4 4 6 6 Soy isoflavones 10 10 min min 8 8 流速3倍、 グラジエント条件最適化 triple flow rate and optimizing the gradient condition YMC-UltraHT Hydrosphere C18 2μm 50×2.0 mmI.D. YMC-UltraHT Hydrosphere C18 50×2.0 mmI.D. 2 µm 3 3 1 1 2 2 Rs(7,8)=1.75 5 4 5 4 6 6 7 8 7 8 Rs(10,11)=2.64 Rs(10,11)=2.64 9 9 10 10 12 12 11 11 12.5-30% B (0-3.3 min) 0.9 mL/minACN (0-3.3 min) 12.5-30% 0.6 µL injection 0.9 mL/min 1. Daidzin 2. Glycitin 3. Genistin 4. 6"-O-Malonyldaidzin 5. 6"-O-Malonylglycitin 6. 6"-O-Acetyldaidzin 7. 6"-O-Acetylglycitin 8. 6"-O-Malonylgenistin 9. Daidzein 10. Glycitein 11. 6"-O-Acetylgenistin 12. Genistein MPa (N070130L) 0 0 (N070130L) 1 1 2 2 3 3 3.3 min 3.3 min Eluent :A) water/acetic acid (100/3) B) acetonitrile/acetic acid (100/3) Temperature:35 ℃ Detection :UV at 254 nm Ordering information Analytical columns Particle size (µm) S-2 Pore size Column size (Å) inner diameter×length(mm) 120 2.0× 30 2.0× 50 2.0× 75 2.0×100 3.0× 50 3.0× 75 3.0×100 Product number HS12S02-0302WT HS12S02-0502WT HS12S02-L502WT HS12S02-1002WT HS12S02-0503WT HS12S02-L503WT HS12S02-1003WT 27 第01章.indd 27 11.1.28 10:40:30 AM High performance reversed-phase column Features of Pro series ODS columns Pro series high performance reversed-phase columns We provide products with excellent performance and feature advanced endcapping technology and quality stable quality all over the world to fulfill the needs of the assurance which is ready for validation. The Pro series pharmaceutical industry, in which production, as well as includes three kinds of ODS, each with unique separation research and development, are increasingly globalized. characteristics. ( ODS ) Reversed-Phase C18 Columns 1 Providing products with stable quality all over the world Feature ● Silica gel base, low in metal impurities ● Processed with advanced endcapping technology ● Superior separation of basic compounds ●U seful for LC/MS ●E xcellent durability ●E xcellent reproducibility ● Two types of test reports are included with each column Standard ODS column with high versatility Low carbon ODS column, useful for separation of hydrophilic compounds High carbon ODS column, useful for separation of highly hydrophobic compounds YMC-Pack Pro C18 Hydrosphere C18 YMC-Pack Pro C18 RS Hydrosphere C18 is an ODS column YMC-Pack performance monomeric type ODS YMC-Pack C18 is a high designed to maintain maximum carbon "polymeric type" ODS bonding, C18 RS is a high column providing standard hydrophilicity without activating the characterized by high resolution and hydrophobicity, high resolution, high silica surface. It can be used with high durability. It is applicable to a durability and excellent reproducibility. 100% water eluent, a problematic wide range of compounds. The It is highly versatile and suitable as the eluent for conventional ODS columns. separation of compounds with relatively first choice column in almost all fields, Hydrosphere C18 is useful for high hydrophobicity and those that differ including pharmaceutical products, separating hydrophilic compounds. only slightly in hydrophobicity, such as agricultural chemicals, foods, and structural isomers, is outstanding. It natural products. also has excellent resistance to acid and alkali, making it useful under demanding separation conditions. Types of Pro series ODS Particle Size (µm) Pore Size (Å) Pro C18 Hydrosphere C18 Pro C18 RS 2 , 3 , 5 , 10 2,3,5 3,5 120 120 80 Specific Surface area (m /g) 330 330 510 Carbon Content 16% 12% 22% monomeric monomeric polymeric 2-8 2-8 1 - 10 Standard type Separation for hydrophilic compounds High durability 2 Bonding type Available pH Range Characteristics 28 第01章.indd 28 11.1.28 10:40:32 AM Three kinds of ODS, each with different separation characteristics The three kinds of ODS columns in the Pro series each have unique carbon content and separation characteristics, including hydrophobicity, plane recognition ability, and hydrogen-bonding capacity. Selection according to the analyte and mobile phase of Pro C18 with standard hydrophobicity, Pro C18 RS with high hydrophobicity, or Hydrosphere C18 with low hydrophobicity, facilitates method development. Hydrophobicity and plane recognition ability of Pro series ODS 1 C18 Hydrosphere C18 3 1 3 C18 RS 1 5 2 2 YMC-Pack 5 3 4 4 2 5 4 0 J020805B 5 1. Uracil 2. n-Butylbenzene 3. o-Terphenyl 4. n-Amylbenzene 5. Triphenylene 10 15 20 min 0 5 J020805D 10 15 20 min 0 J020805E 5 10 15 20 25 min ( ODS ) Reversed-Phase C18 Columns YMC-Pack 1 Column size :150×4.6 mmI.D. Eluent :methanol/water (80/20) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm Hydrogen-bonding capacity α (caffeine/benzene) Comparison of hydrophobicity and hydrogen-bonding capacity of various columns 0.180 Hydrophobicity: low Hydrogen-bonding capacity: high 0.160 Develosil C30-UG Hydrosphere C18 0.140 Atlantis T3 CAPCELL PAK C18 AQ 0.120 Hydrophobicity: intermediate Hydrogen-bonding capacity: middle CAPCELL PAK C18 MGIII Gemini C18 Pro C18 Unison UK-C18 Inertsil ODS-4 Hypersil GOLD CAPCELL PAK C18 MGII Gemini NX-C18 SunFire Inertsil ODS-3 XBridge C18 XTerra MS C18 Mightysil RP-18 C18 Eternity-C18 L-column ODS 0.100 0.080 TSKgel ODS-100S Cadenza CD-C18 L-column2 ODS Hydrophobicity: high Hydrogen-bonding capacity: low Develosil ODS-SR Pro C18 RS 0.060 Eluent : MeOH/W (80/20) [amylbenzene], MeOH/W (30/70) [caffeine, benzene] 0.040 2.00 4.00 6.00 8.00 10.00 Hydrophobicity k’ (amylbenzene) 12.00 14.00 (S090805A) 29 第01章.indd 29 11.1.28 10:40:35 AM High performance reversed-phase column Features of Pro series ODS columns Comparison of separation behavior on the three kinds of ODS columns in the Pro series Parabenzoates C18 ( ODS ) Reversed-Phase C18 Columns 1 3 types of ODS are available to separate the para-benzoates shown in the left, but the best separation between peaks 5 and 6 is achieved on Pro C18 RS. 1 1 1. OH 2 2 2 COOCH3 Methyl p-hydroxybenzoate 2. OH 34 3 4 5 COOCH2CH3 Ethyl p-hydroxybenzoate 34 3. CH3 OH COOCH CH3 56 6 Isopropyl p-hydroxybenzoate 5 6 4. OH 1 C18 RS Hydrosphere C18 COO(CH2)2CH3 Propyl p-hydroxybenzoate 5. OH 0 0 20 min 10 (J020827D) (J020827C) 10 20 min 0 (J020827E) 10 20 30 min CH3 COOCH2CH CH3 Isobutyl p-hydroxybenzoate 6. OH Column :150×4.6 mmI.D. Eluent :methanol/water (55/45) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 270 nm COO(CH2)3CH3 Butyl p-hydroxybenzoate Flavonoid glycosides 2 1 3 C18 4 J020828A 1 HO OH 4 J020828F 2 3 4 C18 RS J020828C 0 O OH 3 Hydrosphere C18 1 HO 2 5 10 15 min 1. Rutin 2. Isoquercitrin 3. Quercitrin 4. Quercetin OR O R R R R = rutinose = glucose = rhamnose =H Hydrogen-bonding interactions significantly contribute to the retention of hydrophilic compounds, including glycosides. Therefore, the retention is longer in the Hydrosphere C18, which has strong hydrogen-bonding capacity, than in Pro C18 RS, which has highly hydrophobic packing materials. To separate such hydrophilic compounds, the Hydrosphere C18 is suitable for precise separation of multicomponent samples, while Pro C18 RS and Pro C18 are suitable for rapid separation of simple components. Column :150×4.6 mmI.D. Eluent :acetonitrile/10 mM H3PO4 (25/75) Flow rate :1.0 mL/min Temperature:40 ℃ Detection :UV at 260 nm 30 第01章.indd 30 11.1.28 10:40:39 AM Terphenyl isomers C18 C18 RS Hydrosphere C18 Brand C1 2 2,3 2 1. 1 3 1 o-Terphenyl 3 1 1 3 2. m-Terphenyl 3. 0 J020719J 10 20 min 0 J020722A 10 20 min 0 J020719E 10 20 min 0 J020719K 10 ( ODS ) Reversed-Phase C18 Columns Since Pro C18 RS has excellent hydrophobic selectivity and shape selectivity, it works well in the separation of structural isomers that are not easily separated by other ODS. 2 1 20 min p-Terphenyl Column :150×4.6 mmI.D. Eluent :methanol/water (85/15) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm Saikosaponins Peaks 2 and 3, which are not easily separated by other ODS, can be completely separated by Hydrosphere C18. It is thought that hydrogen-bonding interaction of hydrophilic functional groups in sugar chain and terpene skeleton as well as hydrophobic interaction has great contribution to the separation of saikosaponins. Hydrosphere C18 is the most appropriate in the separation of this kind of class of compound. 1 3 2 C18 4 F030222B 1 Hydrosphere C18 23 1. 4 3. O H CH2OH OH OH F030222A 1 2,3 O O glucose 6 glucose fucose 4 rhamnose C18 RS Saikosaponin c 2. O H H OH OH O 2,3 fucose 4 10 20 30 3 CH2OH glucose Saikosaponin a F030224E 0 4. O 1 Brand B2 CH2OH glucose Saikosaponin b2 4 F030222C 3 40 min O fucose 3 CH2OH glucose Saikosaponin d Column :150×4.6 mmI.D. Eluent :acetonitrile/water (38/62) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 210 nm 31 第01章.indd 31 11.1.28 10:40:44 AM Analytical columns YMC-Pack Pro C18 ●S uperior separation of basic compounds ●P rocessed with advanced ● Excellent reproducibility ● Utilizes highly pure silica gel base ●T wo types of test reports are included endcapping technology ■Pore size:120 Å ■Carbon content:16% ■Usable pH range:2.0 〜 8.0 with each column Perfectly endcapped ODS YMC-Pack Pro C18 includes more advanced endcapping technology for strictly controlled processing of residual silanol groups that are likely to affect quality. Pro C18 is highly appropriate for basic compounds, including pharmaceutical products and agricultural chemicals. ( ODS ) Reversed-Phase C18 Columns 1 Two test reports are included with each product; a Column Test Report, indicating the column performance, and a Packing Material Test Report, detailing strictly controlled physical properties and separation characteristics of the packing material. Useful for separation of basic compounds Tricyclic antidepressants 1. 23 1 2. Advanced endcapping technology results in good peak shapes even for basic compounds that often elute with poor tailing shapes on competitive columns. ・ HCI 4 N CH2CH2CH2NHCH3 Desipramine hydrochloride Acenaphthene(I.S.) 3. 4. ・ HCI ・ HCI N CH2CH2CH2N CH3 CHCH2CH2N CH3 Imipramine hydrochloride 0 10 CH3 CH3 Amitriptyline hydrochloride 20 min (A961015G) Column :YMC-Pack Pro C18 (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :20 mM KH2PO4-K2HPO4 (pH6.9)/ methanol (25/75) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm Basic peptides C18 4 1 5 1. Bradykinin 2. Dynorphin A (Human,1-13) 3. Eledoisin related peptide 4. Neurotensin 5. Bombesin 6. Substance P ODS-A 4 6 1 2 3 5 6 2 3 0 5 10 15 20 min (A990123B) 0 5 10 15 20 min (A990121A) Neutral peptides C18 2 3 ODS-A 5 4 2 6 3 1 0 YMC-Pack Pro C18 is processed using advanced endcapping technology and produces a better chromatogram than that obtained using ODS-A for separating basic peptides. Similar chromatograms have been obtained with YMCPack Pro C18 and ODS-A for separation of neutral peptides. Column :YMC-Pack Pro C18 and ODS-A (5 µm, 120 Å) 75×4.6 mmI.D. Eluent :A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 20-40% B (0-20 min,linear) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 220 nm 5 4 6 1 5 (A990121B) 10 15 20 min 0 5 (A990119B) 10 15 20 min 1. Oxytocin 2. Met-Enkephalin 3. Leu-Enkephalin 4. Angiotensin l 5. α-Mating factor 6. Insulin 32 第01章.indd 32 11.1.28 10:40:48 AM Excellent lot-to-lot reproducibility batch-to-batch Test A Test B 1. Caffeine 2. Pyridine 3. Phenol Lot C Lot C Lot B Lot B Lot A Lot A 0 5 10 15 A three-lot comparison is shown on the left. Excellent reproducibility is achieved in separating hydrophilic, basic and acidic compounds, as well as hydrophobic compounds. ( ODS ) Reversed-Phase C18 Columns 1. Uracil 2. 4-n-Butoxybenzoic acid 3. Ethylbenzene 4. n-Propylbenzene 0 min 4 8 min 1 column-to-column 150×4.6 mml.D. theoretical plate number Advanced packing technology results in high column efficiency. All packed columns are inspected and controlled according to the strictest quality control criteria for column performance ever, including theoretical plate number testing. Theoretical plate number 24000 21000 18000 15000 12000 9000 6000 3000 20 40 60 80 100 Column No. Two types of test reports 〈Packing Material Test Report〉 〈Column Test Report〉 Each YMC-Pack Pro C18 includes both, a Column Test Report and a Packing Material Test Report, to certify lotto-lot and column-to-column reproducibility. This report describes basic physical properties of silica The Column Test Report shows the theoretical plate number gel, including particle size, pore size and metal content, and asymmetry factor to indicate column performance. For carbon loading after modification, and the specifications convenience in deterioration evaluation, this report includes all and results of separation characteristics tests. analytical conditions including sample concentration as well. 33 第01章.indd 33 11.1.28 10:40:51 AM Analytical columns YMC-Pack Pro C18 Application 3 4 5 3 1 C18 5 3 Brand I4 1 2 4, 5 Brand L1 1 2 2 6 0 5 10 6 F040119B 15 0 20 min 5 10 6 F030130C 15 20 min F040130E 0 5 10 15 20 min nasal drops (OTC) 1 2 CHCOOH CHCOOH 3 CH3 O NHCCH2 N(CH2 CH3 )2 CH3 4 H N Maleic acid Lidocaine 5 6 OH COOCH3 CH3 CH3 CH3 Chlorpheniramine CH3 + CH3 C CH2 C CH3 CH3 Methyl p-hydroxybenzoate Cl Naphazoline CH3 N CHCH2CH2N CH2 N ( ODS ) Reversed-Phase C18 Columns 1 4 (OCH2CH2)2 N CH2 CH3 Cl- Column :150×4.6 mmI.D. Eluent :A) 20 mM KH2PO4-H3PO4 (pH 2.5) B) methanol 20-90% B (0-15 min), 90% B (15-20 min) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 260 nm Benzethonium chloride Application (N050620A) catechins 1 3 1. (-)-Epigallocatechin 2. (+)-Catechin 3. (-)-Epicatechin 4. (-)-Epigallocatechin gallate 5. (-)-Epicatechin gallate 4 2 Column : YMC-Pack Pro C18 (5 µm, 120 Å) 150×4.6 mmI.D. Eluent : acetonitrile/ethyl acetate/water/TFA (9/3/88/0.1) Flow rate : 1.0 mL/min Temperature: 37 ℃ Detection : UV at 280 nm 5 0 10 20 min Application (A970526C) antioxidants 5 1. n-Propyl gallate(PG) 2. 2,4,5-Trihydroxybutyrophenone (THBP) 3. tert-Butylhydroquinone(TBHQ) 4. Nordihydroguaiaretic acid (NDGA) 5. 4-Hydroxymethyl-2,6-di-tert-butylphenol (HMBP) 6. n-Octyl gallate(OG) 7. n-Dodecyl gallate(DG) 4 6 1 7 2 Column :YMC-Pack Pro C18 (5 µm, 120 Å) 75×4.6 mm I.D. Eluent :A) acetonitrile/methanol/5% acetic acid (20/20/60) B) acetonitrile/methanol/5% acetic acid (45/45/10) 0-100% B (0-5 min), 100% B (5-15 min) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 280 nm 3 0 5 10 15 min 34 第01章.indd 34 11.1.28 10:40:55 AM Ordering information Analytical columns Particle size (µm) S-3 S-10 Pore size Column size (Å) inner diameter×length(mm) 120 120 120 1.0× 35 1.0× 50 1.0×100 1.5× 35 1.5× 50 1.5×100 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6× 35 4.6× 50 4.6× 75 4.6×100 4.6×150 6.0× 50 6.0× 75 6.0×100 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 2.0×250 3.0× 50 3.0× 75 3.0×100 3.0×150 3.0×250 4.6× 50 4.6× 75 4.6×100 4.6×150 4.6×250 6.0×150 6.0×250 4.6×150 4.6×250 Product number AS12S03-H501WT AS12S03-0501WT AS12S03-1001WT AS12S03-H5P5WT AS12S03-05P5WT AS12S03-10P5WT AS12S03-H502WT AS12S03-0502WT AS12S03-L502WT AS12S03-1002WT AS12S03-1502WT AS12S03-0503WT AS12S03-1003WT AS12S03-1503WT AS12S03-H546WT AS12S03-0546WT AS12S03-L546WT AS12S03-1046WT AS12S03-1546WT AS12S03-0506WT AS12S03-L506WT AS12S03-1006WT AS12S05-1501WT AS12S05-2501WT AS12S05-15P5WT AS12S05-25P5WT AS12S05-H502WT AS12S05-0502WT AS12S05-L502WT AS12S05-1002WT AS12S05-1502WT AS12S05-2502WT AS12S05-0503WT AS12S05-L503WT AS12S05-1003WT AS12S05-1503WT AS12S05-2503WT AS12S05-0546WT AS12S05-L546WT AS12S05-1046WT AS12S05-1546WT AS12S05-2546WT AS12S05-1506WT AS12S05-2506WT AS12S11-1546WT AS12S11-2546WT Particle size (µm) S-5 S-10 Pore size Column size (Å) inner diameter×length(mm) 120 120 10×150 10×250 20×100 20×150 20×250 20×100 20×150 20×250 Product number AS12S05-1510WT AS12S05-2510WT AS12S05-1020WT AS12S05-1520WT AS12S05-2520WT AS12S11-1020WT AS12S11-1520WT AS12S11-2520WT See P142 for preparative columns other than those listed above. Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) AS12S03-0101CC 1.0×10 S-3 120 S-5 120 1.5×10 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 AS12S03-01P5CC AS12S03-0102CC AS12S03-G304CC AS12S05-0101CC AS12S05-01P5CC AS12S05-0102CC AS12S05-G304CC ( ODS ) Reversed-Phase C18 Columns S-5 Semi-preparative columns 1 A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) YMC guard cartridge pack S-3 4.0×23 120 S-5 4.0×23 120 XPCHSMW XPCHW AS12S03-G304CCPK AS12S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-5 120 S-10 120 10×30 20×50 AS12S05-0310WTG AS12S05-0520WTG 20×50 AS12S11-0520WTG When columns packed with material from different lots are needed for validation purposes, please contact us. 35 第01章.indd 35 11.1.28 10:40:56 AM Analytical columns Hydrosphere C18 ● Strong retention of hydrophilic compounds ● Can be used with 100% water mobile phase ● Superior separation of basic compounds ● Utilizes highly pure silica gel base ●T wo types of test reports are included with each column ■Pore size:120 Å ■Carbon content:12% ■Usable pH range:2.0 〜 8.0 ● Excellent reproducibility Hydrophilic ODS Hydrosphere C18 is designed to maintain adequate hydrophilicity on the packing surface for superior separation of hydrophilic compounds. Hydrophilic compounds are retained with much greater affinity than with conventional ODS columns, permitting the product to be used ( ODS ) Reversed-Phase C18 Columns 1 with 100% water eluent, a problematic eluent for conventional ODS columns. Hydrosphere C18 is useful for separating a wide range of compounds, including nucleic acids and their derivatives, organic acids, saccharides, glycosides and peptides. Appropriate for separation of hydrophilic compounds Separation of organic acids conventional ODS column(BrandⅠ) Hydrosphere C18 1 1 3 0 4 3,4 5 5 5 A000310E High hydrophilicity of the Hydrosphere C18 packing material surface enables longer retention and superior separation of citric acid and fumaric acid as compared to a conventional ODS column (Brand Ⅰ). 2 2 10 0 min 5 A000615A 10 Column :150×4.6 mm I.D. Eluent :20 mM NaH2PO4-H3PO4 (pH 2.8) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 210 nm 1. Lactic acid 2. Acetic acid 3. Citric acid 4. Fumaric acid 5. Succinic acid min Reproducibility of retention time when used with 100% water mobile phase conventional ODS column(Brand Ⅲ) conventional ODS column(Brand Ⅱ) Hydrosphere C18 Before standing 3 1 2 3 5※ 10 20 min ※ 0 S0517A 10 5※ 4 0 S0518H After standing S0515A 2 1 1 4 0 3 2 10 ※ 98% 20 min 4 0 S0519A 20 min 0 S0518F 10 ※ 89% 10 20 min 0 S0519C 5※ 10 20 min 71% 20 min After completion of analysis, the flow was stopped and the columns were left to stand overnight (ca. 15 hours). 1. Cytosine 2. Uracil 3. Guanine 4. Thymine 5. Adenine Column :150×4.6 mm I.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm When conventional ODS columns are used with 100% water mobile phase, the apparent hydrophobicity decreases due to repulsive interactions between water and hydrophobic groups on the surface of the packing materials, reducing retention of compounds. In the experiment illustrated here, columns filled with 100% water mobile phase were left to stand overnight. Retention times of nucleic acid bases were analyzed before and after standing and compared. The retention times of adenine after standing decreased to 89% of the original value for Brand II and to 71% of the original Brand III value. By contrast, Hydrosphere C18 retained 98% of its original retention for adenine. Hydrosphere C18 is designed to maintain adequate hydrophilicity on the packing surface so that hydration can be achieved, and when Hydrosphere C18 is used with mobile phase containing no organic solvent, the retention time is not significantly shortened and highly reproducible chromatograms can be obtained. 36 第01章.indd 36 11.1.28 10:41:01 AM Differences in separation characteristics from conventional ODS columns Hydrosphere C18 2 3 0 10 S000309A 4 5 20 6 30 40 min C18 2 1 4 3 0 10 S990607B 5 20 30 6 40 min ( ODS ) Reversed-Phase C18 Columns The chromatograms shown were obtained in tests conducted to compare separation characteristics using a mixed sample containing compounds with different characteristics. The packing material surface of Hydrosphere C18 has higher hydrophilicity and lower C18 ligand density than those of Pro C18. This results in shorter retention of hydrophobic compounds, including n-propylbenzene and n-butylbenzene, and planar compounds like acenaphthene. By contrast, retention of testosterone, a compound with large hydrogen-bonding properties, is longer on Hydrosphere C18 and on Brand IV, which also has a highly hydrophilic packing surface. Amitriptyline, a basic compound, caused tailing peaks in Brand IV and Brand V. By contrast, Hydrosphere C18 and Pro C18 produce good peak shapes for such basic compounds. Although the retention times of amitriptyline on Hydrosphere C18 and Pro 18 are similar, there is a big difference between the two columns with regard to resolution of n-butylbenzene and amitriptyline. This difference is considered to be due to the hydrophilicity of the Hydrosphere C18 surface. These results demonstrate that Hydrosphere C18, having different separation characteristics from conventional ODS columns, enables separation of compounds that cannot be separated using other ODS columns. 1 1 Brand Ⅳ 1 2 4 3 5 6 0 10 F000614B 20 30 40 50 60 min Brand Ⅴ 2 1 3 4 5 6 0 F000614C 10 20 30 40 50 1. Toluene 2. Testosterone 3. n-Propylbenzene 4. Acenaphthene 5. n-Butylbenzene 6. Amitriptyline hydrochloride 60 70 80 90 min Column :150×4.6 mm I.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)/methanol (35/65) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm 37 第01章.indd 37 11.1.28 10:41:04 AM Analytical columns Hydrosphere C18 Short analysis time conventional ODS column Hydrosphere C18 3 2 4 2 4 ( ODS ) Reversed-Phase C18 Columns 1 Hydrosphere C18 can reduce the time necessary for analyzing compounds with high hydrophobicity by about 20% as compared with conventional ODS columns. 3 5 6 1 5 6 Antioxidants 1 1. Nordihydroguaiaretic acid (NDGA) 2. Butyl hydroxyanisol (BHA) 3. 4-Hydroxy-2,6-di-tert-butylphenol (HMBP) 4. n-Octyl gallate (OG) 5. Dibutyl hydroxytoluene (BHT) 6. n-Dodesyl gallate (DG) 0 J010216B 5 10 15 0 min J010215B 5 10 15 min Column :50×4.6 mm I.D. (3 µm, 120 Å) Eluent :acetonitrile/methanol/5% acetic acid (35/35/30) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 280 nm Application (F000707B) Nicotinic acid and derivatives 4 1 2 1. Nicotinamide adenine dinucleotide phosphate oxidized form (NADP) 2. Nicotinic acid 3. Nicotinamide adenine dinucleotide phosphate reduced form (NADPH) 4. Nicotinamide 5. β-Nicotinamide adenine dinucleotide (NAD) 6. β-Nicotinamide adenine dinucleotide reduced form (NADH) 5 Column :Hydrosphere C18 (5 µm, 120 Å) 150×4.6 mm I.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)/acetonitrile (99/1) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 260 nm 3 6 0 5 10 15 20 25 min Application (A000922A) Nucleotides 5 1. 5’-UTP 2. 5’-UDP 3. 5’-CMP 4. 5’-UMP 5. 5’-dCMP 6. 5’-GMP 1 7 2 4 8 0 10 Column :Hydrosphere C18 (5 µm, 120 Å) 150×4.6 mm I.D. Eluent :100 mM KH2PO4-K2HPO4 (pH 5.5) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 260 nm 11 6 3 9 5 10 15 7. 5’-ATP 8. 5’-IMP 9. 5’-ADP 10. 5’-TMP 11. 5’-AMP 20 min 38 第01章.indd 38 11.1.28 10:41:07 AM Ordering information Analytical columns Particle size (µm) S-3 Pore size Column size (Å) inner diameter×length(mm) 120 120 1.0× 35 1.0× 50 1.0×100 1.5× 35 1.5× 50 1.5×100 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6× 35 4.6× 50 4.6× 75 4.6×100 4.6×150 6.0× 50 6.0× 75 6.0×100 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 2.0×250 3.0× 50 3.0× 75 3.0×100 3.0×150 3.0×250 4.6× 50 4.6× 75 4.6×100 4.6×150 4.6×250 6.0×150 6.0×250 Product number HS12S03-H501WT HS12S03-0501WT HS12S03-1001WT HS12S03-H5P5WT HS12S03-05P5WT HS12S03-10P5WT HS12S03-H502WT HS12S03-0502WT HS12S03-L502WT HS12S03-1002WT HS12S03-1502WT HS12S03-0503WT HS12S03-1003WT HS12S03-1503WT HS12S03-H546WT HS12S03-0546WT HS12S03-L546WT HS12S03-1046WT HS12S03-1546WT HS12S03-0506WT HS12S03-L506WT HS12S03-1006WT HS12S05-1501WT HS12S05-2501WT HS12S05-15P5WT HS12S05-25P5WT HS12S05-H502WT HS12S05-0502WT HS12S05-L502WT HS12S05-1002WT HS12S05-1502WT HS12S05-2502WT HS12S05-0503WT HS12S05-L503WT HS12S05-1003WT HS12S05-1503WT HS12S05-2503WT HS12S05-0546WT HS12S05-L546WT HS12S05-1046WT HS12S05-1546WT HS12S05-2546WT HS12S05-1506WT HS12S05-2506WT Particle size (µm) S-5 Pore size Column size (Å) inner diameter×length(mm) 120 10×150 10×250 20×100 20×150 20×250 Product number HS12S05-1510WT HS12S05-2510WT HS12S05-1020WT HS12S05-1520WT HS12S05-2520WT Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) HS12S03-0101CC 1.0×10 S-3 120 S-5 120 1.5×10 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 HS12S03-01P5CC HS12S03-0102CC HS12S03-G304CC HS12S05-0101CC HS12S05-01P5CC HS12S05-0102CC HS12S05-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) YMC guard cartridge pack S-3 4.0×23 120 S-5 4.0×23 120 ( ODS ) Reversed-Phase C18 Columns S-5 Semi-preparative columns 1 XPCHSMW XPCHW HS12S03-G304CCPK HS12S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-5 120 10×30 20×50 HS12S05-0310WTG HS12S05-0520WTG 39 第01章.indd 39 11.1.28 10:41:08 AM Analytical columns YMC-Pack Pro C18 RS ●E xcellent acid resistance and alkali resistance (pH 1 to 10) ● Superior separation of structural isomers ● Excellent reproducibility ● Utilizes highly pure silica gel base ●T wo types of test reports are included ● Superior separation of basic compounds ■Pore size:80 Å ■Carbon content:22% ■Usable pH range:1.0 〜 10.0 with each column Highly durable ODS YMC-Pack Pro C18 RS is a polymeric type high carbon ODS column characterized by high resolution and high durability. It is applicable to a wide range of compounds, providing good separation of basic compounds that easily cause Excellent durability The pH 1 C18 RS packing material hydrophobicity is hardly changed Unlike conventional ODS columns, pH 10 . 100 100 80 80 60 60 % 20 0 40 ( ) YMC-Pack Pro C18 RS Acid-resistant column (manufactured by other company) Monomeric ODS 40 0 C18 RS can be used with an alkali mobile phase. under the harsh conditions of pH1 and column temperature 70 ℃. % ( ODS ) Reversed-Phase C18 Columns 1 tailing peaks. The separation selectivity for compounds that differ only slightly in hydrophobicity is outstanding. It also has excellent resistance to acid and alkali, making it useful under demanding separation conditions. Pro C18 RS Monomeric ODS 20 0 20 40 60 80 100(hours) Hours exposure to acetonitrile/water/TFA (10/90/1), change in packing material hydrophobicity at 70 ℃. 0 200 400 600 800(hours) Sequential analysis using 20 mM H3BO3-NaOH (pH 9.8)/methanol (50/50) A wide choice of mobile phases facilitates method development 150×4.6 mmI.D. 5 µm pH 3.2 3,4,5 50×4.6 mmI.D. 3 µm pH 6.8 pH 8.8 pH 8.8 Downsizing 1 2 12 2 3 F040716A 5 10 1. Flurbiprofen 2. Methyl benzoate 3. Lidocaine 4. Diltiazem 5. Verapamil 15 4 3 4 5 5 1 0 12 3 4 5 20 min 0 F040714E 5 10 15 20 min Column :YMC-Pack Pro C18 RS Eluent :20 mM acetate buffer/methanol (35/65) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 220 nm 0 F040714C 5 10 15 20 min 0 F040714D 5 10 min Pro C18 RS with excellent acid and alkali resistance can be used over a wide pH range, and it is easy to optimize the separating conditions. In addition, by downsizing from the column (particle 5 µm, length 150 mm) to a shorter column (particle 3 µm, length 50 mm), with high separation ability, it becomes possible to shorten the analysis time by one third without affecting the separation. 40 第01章.indd 40 11.1.28 10:41:12 AM Separation of basic compounds YMC-Pack C18 RS Acid-resistant column manufactured by other company(Brand D3) 4 Tf(4) 2.87 3 4 1 0 10 J011114B 20 min 0 10 J020523D 20 min 1. Desipramine 2. Diphenyl (I.S.) 3. Imipramine 4. Amitriptyline Column :150×4.6 mmI.D. Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9) /methanol (25/75) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm Separation of structural isomers Pro C18 RS with a high C18 density is excellent in hydrophobicity recognition ability and plane recognition ability, and can be applied to separate structural isomers which are difficult to separate with other ODS. In addition, by using 3 µm column packing, resolution can be increased, and it becomes possible to achieve a baseline separation of linolenate isomers. C18 RS 5 µm Brand F1 (standard-type ODS 5 µm) 1 1 2 2 ( ODS ) Reversed-Phase C18 Columns 3 1 2 Tf(4) 1.16 2 Amitriptyline is a basic compound that can easily tail on some silica based columns. In comparison with competing acid-resistant columns, Pro C18 RS produces good peak shapes without tailing. 1 Rs=1.24 Rs=0.8 0 F040713D 10 20 30 min 0 F040714 10 20 30 min Methyl linolenate isomers 1. Methyl linolenate 2. Methyl γ-linolenate Column :150×4.6 mmI.D. Eluent :acetonitrile/THF/water (35/30/35) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 210 nm C18 RS 3 µm 1 2 Rs=1.54 0 F040713A 10 20 30 min 41 第01章.indd 41 11.1.28 10:41:15 AM Analytical columns YMC-Pack Pro C18 RS Separation of peptides by LC/MS 530.95 M+2 100 % ( ODS ) Reversed-Phase C18 Columns 1 Pro C18 RS produces good peak shapes when it is used with mobile phase containing low TFA concentration, making it suitable for LC/ MS. Bradykinin(MW: 1059.56) 531.47 M+1 0 1060.27 400 600 800 2 100 1 % 18 5.00 1000 3 5 1200(m/z) 1. [Arg8]-Vasopressin 2. Bradykinin 3. Angiotensin Ⅱ 4. Oxytocin 5. Angiotensin Ⅰ 6. Leu-Enkephalin 6 4 10.00 15.00 Column :YMC-Pack Pro C18 RS (5 µm, 80 Å) 150×2.0 mmI.D. Eluent :A) water/TFA (100/0.01) B) acetonitrile/TFA (100/0.01) 10-35% B (0-15 min) Flow rate :0.2 mL/min Temperature:37 ℃ Detection :ESI positive-mode 20.00 (Time) Application Chloroacetophenone isomers 1. 2 Chlorophenol isomers COCH3 Cl 1. o-Chloroacetophenone 2. 3. 1 2. 3 J011026B 10 20 min 3. 1 Cl m-Chloroacetophenone 0 OH Cl p-Chlorophenol COCH3 Column :YMC-Pack Pro C18 RS (5 µm, 80 Å) 150×4.6 mmI.D. Eluent :acetonitrile/water (40/60) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 254 nm Cl o-Chlorophenol COCH3 Cl p-Chloroacetophenone 3 OH 2 OH Cl m-Chlorophenol Column Eluent 0 J011113D 10 20 min :YMC-Pack Pro C18 RS (5 µm, 80 Å) 150×4.6 mmI.D. :a cetonitrile/water/acetic acid (30/70/1) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 230 nm 42 第01章.indd 42 11.1.28 10:41:19 AM Ordering information Analytical columns Particle size (µm) S-3 Pore size Column size (Å) inner diameter×length(mm) 80 80 1.0× 35 1.0× 50 1.0×100 1.5× 35 1.5× 50 1.5×100 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6× 35 4.6× 50 4.6× 75 4.6×100 4.6×150 6.0× 50 6.0× 75 6.0×100 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 2.0×250 3.0× 50 3.0× 75 3.0×100 3.0×150 3.0×250 4.6× 50 4.6× 75 4.6×100 4.6×150 4.6×250 6.0×150 6.0×250 Product number RS08S03-H501WT RS08S03-0501WT RS08S03-1001WT RS08S03-H5P5WT RS08S03-05P5WT RS08S03-10P5WT RS08S03-H502WT RS08S03-0502WT RS08S03-L502WT RS08S03-1002WT RS08S03-1502WT RS08S03-0503WT RS08S03-1003WT RS08S03-1503WT RS08S03-H546WT RS08S03-0546WT RS08S03-L546WT RS08S03-1046WT RS08S03-1546WT RS08S03-0506WT RS08S03-L506WT RS08S03-1006WT RS08S05-1501WT RS08S05-2501WT RS08S05-15P5WT RS08S05-25P5WT RS08S05-H502WT RS08S05-0502WT RS08S05-L502WT RS08S05-1002WT RS08S05-1502WT RS08S05-2502WT RS08S05-0503WT RS08S05-L503WT RS08S05-1003WT RS08S05-1503WT RS08S05-2503WT RS08S05-0546WT RS08S05-L546WT RS08S05-1046WT RS08S05-1546WT RS08S05-2546WT RS08S05-1506WT RS08S05-2506WT Particle size (µm) S-5 Pore size Column size (Å) inner diameter×length(mm) 80 10×150 10×250 20×100 20×150 20×250 Product number RS08S05-1510WT RS08S05-2510WT RS08S05-1020WT RS08S05-1520WT RS08S05-2520WT Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) RS08S03-0101CC 1.0×10 S-3 80 S-5 80 1.5×10 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 RS08S03-01P5CC RS08S03-0102CC RS08S03-G304CC RS08S05-0101CC RS08S05-01P5CC RS08S05-0102CC RS08S05-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) YMC guard cartridge pack S-3 80 4.0×23 S-5 80 4.0×23 ( ODS ) Reversed-Phase C18 Columns S-5 Semi-preparative columns 1 XPCHSMW XPCHW RS08S03-G304CCPK RS08S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-5 80 10×30 20×50 RS08S05-0310WTG RS08S05-0520WTG 43 第01章.indd 43 11.1.28 10:41:20 AM Analytical columns YMC-Pack ODS-A ● Conventional ODS column ● Currently in use worldwide ■Pore size:120 Å, 200 Å, 300 Å ■Carbon content:17% (120 Å), 12% (200 Å), 7% (300 Å) ■Usable pH range:2.0 〜 7.5 Standard ODS YMC-Pack ODS-A has a highly endcapped surface structure and appropriate hydrophobicity for separation of a wide range of compounds. It is produced under strict quality control with respect to 50 or more parameters in Quality control system for excellent performance The graph indicates lot-to-lot reproducibility with respect to hydrophobic interaction for 10 lots, an essential column performance characteristic. Extremely stable lot-to-lot reproducibility of the hydrophobic surface is achieved by strict control of variables. In addition to measurement of the physical properties, a wide range of compounds including hydrophobic, acidic, basic and coordination compounds are analyzed under optimal conditions to evaluate column performance. 10 tR (Butyl benzoate), min ( ODS ) Reversed-Phase C18 Columns 1 order to ensure stable quality. This product is highly regarded as the standard YMCPack packing material in various countries around the world. 8 6 4 2 0 1 2 3 4 5 6 7 Lot Number 8 9 10 Application (S930422D) Application Analgesics O 1 CH3COHN OH 2 3 3 OC2H5 Ethenzamide CH3 N N N CH3 1.α-Carotene 2.β-Carotene 2 N Caffeine 4 CONH2 H 3C O Acetaminophen 1 Carotenes 1 2 CH2= CHCH2 H 3C H 3C CHCHCONHCONH 2 Apronalide 4 0 10 20 30 min (G910523Q) Column :YMC-Pack ODS-A (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :methanol/water (40/60) Flow rate :0.7 mL/min Temperature:30 ℃ Detection :UV at 220 nm 0 10 20 min Column :YMC-Pack ODS-A (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :acetonitrile/methanol/THF (50/45/5) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 450 nm Ordering information Analytical columns Particle size (µm) S-3 Pore size Column size (Å) inner diameter×length(mm) 120 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 Product number AA12S03-H502WT AA12S03-0502WT AA12S03-L502WT AA12S03-1002WT AA12S03-1502WT AA12S03-0503WT AA12S03-1003WT AA12S03-1503WT Particle size (µm) S-3 Pore size Column size (Å) inner diameter×length(mm) 120 4.6× 75 4.6×100 4.6×150 6.0×100 6.0×150 Product number AA12S03-L546WT AA12S03-1046WT AA12S03-1546WT AA12S03-1006WT AA12S03-1506WT 44 第01章.indd 44 11.1.28 10:41:23 AM Ordering information Semi-preparative columns Analytical columns Particle size (µm) S-5 Pore size Column size (Å) inner diameter×length(mm) 120 300 S-10 120 300 AA12S05-1501WT AA12S05-2501WT AA12S05-15P5WT AA12S05-25P5WT AA12S05-L502WT AA12S05-1502WT AA12S05-2502WT AA12S05-1503WT AA12S05-2503WT AA12S05-L546WT AA12S05-1046WT AA12S05-1546WT AA12S05-2546WT AA12S05-3046WT AA12S05-1006WT AA12S05-1506WT AA12S05-2506WT AA12S05-3006WT AA20S05-1546WT AA20S05-2546WT AA30S05-1501WT AA30S05-2501WT AA30S05-15P5WT AA30S05-25P5WT AA30S05-L502WT AA30S05-1502WT AA30S05-2502WT AA30S05-L546WT AA30S05-1046WT AA30S05-1546WT AA30S05-2546WT AA30S05-3046WT AA30S05-1006WT AA30S05-1506WT AA30S05-2506WT AA30S05-3006WT AA12S11-1046WT AA12S11-1546WT AA12S11-2546WT AA12S11-3046WT AA12S11-1006WT AA12S11-1506WT AA12S11-2506WT AA12S11-3006WT AA30S11-1046WT AA30S11-1546WT AA30S11-2546WT AA30S11-3046WT AA30S11-1006WT AA30S11-1506WT AA30S11-2506WT AA30S11-3006WT Particle size (µm) S-10 S-5 Pore size Column size (Å) inner diameter×length(mm) 120 200 300 10×150 10×250 10×300 20×100 20×150 20×250 20×150 20×250 10×150 10×250 10×300 20×150 20×250 120 200 300 AA12S11-1510WT AA12S11-2510WT AA12S11-3010WT AA12S11-1520WT AA12S11-2520WT AA20S11-1520WT AA20S11-2520WT AA30S11-1510WT AA30S11-2510WT AA30S11-3010WT AA30S11-1520WT AA30S11-2520WT Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) AA12S03-0102CC 2.0×10 S-3 S-5 120 200 300 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 AA12S03-G304CC AA12S05-0101CC AA12S05-01P5CC AA12S05-0102CC AA12S05-G304CC AA20S05-G304CC AA30S05-0101CC AA30S05-01P5CC AA30S05-0102CC AA30S05-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) 1 XPCHSMW XPCHW YMC guard cartridge pack S-3 120 4.0×23 AA12S03-G304CCPK S-5 120 4.0×23 AA12S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-3 120 S-5 120 200 Product number AA12S05-1510WT AA12S05-2510WT AA12S05-3010WT AA12S05-1020WT AA12S05-1520WT AA12S05-2520WT AA20S05-1520WT AA20S05-2520WT AA30S05-1510WT AA30S05-2510WT AA30S05-3010WT AA30S05-1520WT AA30S05-2520WT 10×150 10×250 10×300 20×150 20×250 20×150 20×250 10×150 10×250 10×300 20×150 20×250 Product number See P145 for preparative columns other than those listed above. Semi-preparative columns Particle size (µm) Pore size Column size (Å) inner diameter×length(mm) ( ODS ) Reversed-Phase C18 Columns 200 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 4.6×150 4.6×250 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×150 2.0×250 4.6× 75 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 Product number 300 S-10 120 200 300 4.0×10 5.0×10 4.0×10 5.0×10 10×30 20×50 4.0×10 20×50 4.0×10 5.0×10 10×30 20×50 4.0×10 5.0×10 10×30 20×50 4.0×10 5.0×10 20×50 4.0×10 5.0×10 10×30 20×50 AA12S03-0104WFG AA12S03-0105WFG AA12S05-0104WFG AA12S05-0105WFG AA12S05-0310WTG AA12S05-0520WTG AA20S05-0104WFG AA20S05-0520WTG AA30S05-0104WFG AA30S05-0105WFG AA30S05-0310WTG AA30S05-0520WTG AA12S11-0104WFG AA12S11-0105WFG AA12S11-0310WTG AA12S11-0520WTG AA20S11-0104WFG AA20S11-0105WFG AA20S11-0520WTG AA30S11-0104WFG AA30S11-0105WFG AA30S11-0310WTG AA30S11-0520WTG 45 第01章.indd 45 11.1.28 10:41:24 AM Analytical columns YMC-Pack ODS-AM ● Similar to ODS-A in selectivity ● Excellent reproducibility ● Useful for quality control purposes ■Pore size:120 Å ■Carbon content:17% ■Usable pH range:2.0 〜 7.5 ODS with outstanding lot-to-lot reproducibility YMC-Pack ODS-AM is a product which especially emphasizes lot-to-lot reproducibility of the packing material. Strict control is performed on all aspects of quality, including physical properties of silica gel base Special attention is given to lot-to-lot reproducibility 10 α (Methylparben/2,6-Dimethylpyridine) ( ODS ) Reversed-Phase C18 Columns 1 and surface modifying processes, in order to ensure stable quality. Double endcapping is conducted to ensure that lot-to-lot reproducibility is maintained. The separation factor (α) of methylparaben/2,6dimethylpyridine for 10 lots of ODS-AM packing material is plotted on the left graph. Excellent lot-to-lot reproducibility is achieved even in separation of basic compounds. 8 6 4 2 0 1 2 3 4 5 6 7 8 9 10 Lot Number Acidic compounds Coordination compounds 1. Uracil 2. Benzoic acid 3. p-Toluic acid 4. p-Ethylbenzoic acid 5. Methy benzoate 1 1. Uracil 2. Acetylacetone 3. 8-Quinolinol ODS-AM 3 2 3 ODS with a high degree of metal impurities 1 1 2 4 5 2 3 Secondary interaction of the stationary phase surface may affect peak shapes of acidic compounds. ODS-AM can provide chromatograms without tailing peaks as shown above. The peak shapes of coordination compounds, including 8-quinolinol and acetylacetone, may be affected by the presence of metal impurities on the surface of the silica. The distribution level of metal impurities on the column is strictly controlled for ODS-AM, providing excellent peak symmetry. 46 第01章.indd 46 11.1.28 10:41:26 AM Application (G910523J) Alkaloids 1 4 3 2 7 5 9 8 6 0 20 40 60 80 min Column :YMC-Pack ODS-AM (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :acetonitrile/20 mM KH2PO4 (20/80) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 210 nm Ordering information Analytical columns Particle size (µm) S-3 S-5 Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) Pore size Column size (Å) inner diameter×length(mm) 120 120 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6× 75 4.6×100 4.6×150 6.0×100 6.0×150 6.0×250 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×100 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 Semi-preparative columns 10×150 S-5 120 10×250 10×300 20×100 20×150 20×250 Product number AM12S03-H502WT AM12S03-0502WT AM12S03-L502WT AM12S03-1002WT AM12S03-1502WT AM12S03-0503WT AM12S03-1003WT AM12S03-1503WT AM12S03-L546WT AM12S03-1046WT AM12S03-1546WT AM12S03-1006WT AM12S03-1506WT AM12S03-2506WT AM12S05-1501WT AM12S05-2501WT AM12S05-15P5WT AM12S05-25P5WT AM12S05-L502WT AM12S05-1002WT AM12S05-1502WT AM12S05-2502WT AM12S05-1503WT AM12S05-2503WT AM12S05-L546WT AM12S05-1046WT AM12S05-1546WT AM12S05-2546WT AM12S05-3046WT AM12S05-1006WT AM12S05-1506WT AM12S05-2506WT AM12S05-3006WT AM12S05-1510WT AM12S05-2510WT AM12S05-3010WT AM12S05-1020WT AM12S05-1520WT AM12S05-2520WT See P147 for preparative columns other than those listed above. 第01章.indd 47 Particle size (µm) Pore size Column size (Å) inner diameter×length(mm) S-3 120 S-5 120 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 ( ODS ) Reversed-Phase C18 Columns 1. Hordenine sulfate 2. Pilocarpine chloride 3. Scopolamine bromide 4. Atropine sulfate 5. Cinchonine 6. Sparteine sulfate 7. Quinine 8. Papaverine chloride 9. Narcotine chloride 1 Product number AM12S03-0102CC AM12S03-G304CC AM12S05-0101CC AM12S05-01P5CC AM12S05-0102CC AM12S05-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) XPCHSMW XPCHW YMC guard cartridge pack S-3 120 4.0×23 AM12S03-G304CCPK S-5 120 4.0×23 AM12S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-3 120 S-5 120 4.0×10 5.0×10 4.0×10 5.0×10 10×30 20×50 AM12S03-0104WFG AM12S03-0105WFG AM12S05-0104WFG AM12S05-0105WFG AM12S05-0310WTG AM12S05-0520WTG 47 11.1.28 10:41:27 AM Analytical columns YMC-Pack ODS-AQ ● Useful for separation of hydrophilic compounds ● Can be used with 100% water in mobile phase ● Different selectivity from conventional ODS ■Pore size:120 Å, 200 Å ■Carbon content:14% , 10% ■Usable pH range:2.0 〜 7.5 Hydrophilic ODS YMC-Pack ODS-AQ has moderate hydrophobicity and hydrogen-bonding capacity. It shows different retention behavior from that of YMC-Pack ODS-A, for samples with relatively high hydrophilicity. ( ODS ) Reversed-Phase C18 Columns 1 It is useful in fields including carbohydrate chemistry for oligosaccharides and glycosides, pharmacognosy and natural product chemistry. Useful for separation of sugars This product is useful for separations using mobile phases in which water content is relatively high, such as separation of sugars and glycosides. G11 G12 G13 1. Maltoundecaose(G11) 2. Maltododecaose(G12) 3. Maltotridecaose(G13) 4. Maltotetradecaose(G14) 5. Maltopentadecaose(G15) G14 G15 10 20 30 Column :YMC-Pack ODS-AQ (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :methanol/water (5/995) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :RI, 8×10-6RIU/FS min (M910604M) Hydrogen-bonding capacity ODS-A ODS-AQ ODS-AL 1 1 1 2 2 3 3 4 4 5 Chromatograms of purine alkaloids are shown on the left. As compared with ODS-A, ODS-AQ and ODS-AL show longer retention of caffeine and theophylline relative to phenol, an internal standard. 2 5 3 5 4 1. Trigonelline 2. Theobromine 3. Theophylline 4. Caffeine 5. Phenol α (Phenol/Caffeine) 1.8 ODS-A 1.6 1.4 ODS-AQ 1.2 ODS-AL Change of separation factor (α) of caffeine/phenol according to temperature change in ODS-AQ, ODS-A and ODS-AL is plotted on the graph shown on the left. Although caffeine is considered to have stronger hydrogenbonding properties than phenol, the hydrogen-bonding capacity of stationary phase may affect the α of caffeine/ phenol. The smaller the hydrogen-bonding capacities are, the higher α of phenol/caffeine. ODS-AQ shows different retention behavior from that of ODS-A since it has moderate hydrogen-bonding capacity. 1.0 3.0 (60 ℃) 3.1 3.2 3 10 /T,K 3.3 -1 3.4 (20 ℃) 48 第01章.indd 48 11.1.28 10:41:30 AM Application (G910607E) Crude drugs (glycosides) 1 1. Arbutin 2. Salicin 3. Esculine 4. G-Strophantine(Ouabain) 5. Baicalin 6. Barbaloin 7. Glycyrrhizin 8. Digitoxine 5 34 2 6 7 8 10 20 30 40 min Ordering information Analytical columns Particle size (µm) S-3 S-5 Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) Pore size Column size (Å) inner diameter×length(mm) 120 120 200 2.0× 35 2.0× 50 2.0× 75 2.0×100 2.0×150 3.0× 50 3.0×100 3.0×150 4.6×100 4.6×150 6.0×100 6.0×150 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 4.6×150 4.6×250 Semi-preparative columns 10×150 S-5 120 200 S-10 120 200 10×250 10×300 20×100 20×150 20×250 20×150 20×250 10×150 10×250 10×300 20×150 20×250 20×150 20×250 Product number AQ12S03-H502WT AQ12S03-0502WT AQ12S03-L502WT AQ12S03-1002WT AQ12S03-1502WT AQ12S03-0503WT AQ12S03-1003WT AQ12S03-1503WT AQ12S03-1046WT AQ12S03-1546WT AQ12S03-1006WT AQ12S03-1506WT AQ12S05-1501WT AQ12S05-2501WT AQ12S05-15P5WT AQ12S05-25P5WT AQ12S05-L502WT AQ12S05-1502WT AQ12S05-2502WT AQ12S05-1503WT AQ12S05-2503WT AQ12S05-L546WT AQ12S05-1046WT AQ12S05-1546WT AQ12S05-2546WT AQ12S05-3046WT AQ12S05-1006WT AQ12S05-1506WT AQ12S05-2506WT AQ12S05-3006WT AQ20S05-1546WT AQ20S05-2546WT Particle size (µm) S-3 120 S-5 120 200 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 4.0×23 Product number AQ12S03-0102CC AQ12S03-G304CC AQ12S05-0101CC AQ12S05-01P5CC AQ12S05-0102CC AQ12S05-G304CC AQ20S05-G304CC 1 A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) XPCHSMW XPCHW YMC guard cartridge pack S-3 120 4.0×23 AQ12S03-G304CCPK S-5 120 4.0×23 AQ12S05-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns S-3 120 S-5 AQ12S05-1510WT AQ12S05-2510WT AQ12S05-3010WT AQ12S05-1020WT AQ12S05-1520WT AQ12S05-2520WT AQ20S05-1520WT AQ20S05-2520WT AQ12S11-1510WT AQ12S11-2510WT AQ12S11-3010WT AQ12S11-1520WT AQ12S11-2520WT AQ20S11-1520WT AQ20S11-2520WT Pore size Column size (Å) inner diameter×length(mm) ( ODS ) Reversed-Phase C18 Columns 0 Column :YMC-Pack ODS-AQ (5 µm, 120 Å) 250×4.6 mmI.D. Eluent :A) methanol/20 mM NH4H2PO4 (5/95) B) methanol/20 mM NH4H2PO4 (80/20) 0-100% B (0-20 min,linear),100% B (20-40 min) Flow rate :0.6 mL/min Temperature:30 ℃ Detection :UV at 250 nm 120 200 S-10 120 200 4.0×10 5.0×10 4.0×10 5.0×10 10×30 20×50 4.0×10 20×50 10×30 20×50 20×50 AQ12S03-0104WFG AQ12S03-0105WFG AQ12S05-0104WFG AQ12S05-0105WFG AQ12S05-0310WTG AQ12S05-0520WTG AQ20S05-0104WFG AQ20S05-0520WTG AQ12S11-0310WTG AQ12S11-0520WTG AQ20S11-0520WTG See P146 for preparative columns other than those listed above. 49 第01章.indd 49 11.1.28 10:41:32 AM Analytical columns YMC-Pack ODS-AL ● ODS with residual silanol groups ● Utilizes secondary interaction caused by silanol groups ■Pore size:120 Å ■Carbon content:17% ■Usable pH range:2.0 〜 7.5 Non-endcapped ODS YMC-Pack ODS-AL uses not only hydrophobic interaction but also secondary interaction caused by silanol groups that affect separation. This results in a different selectivity from conventional ODS columns. ( ODS ) Reversed-Phase C18 Columns 1 When ionic interaction is utilized, it is preferable to use a buffer in the mobile phase to achieve reproducibility of chromatograms. Effects of silanol group ODS-AL The separation factor (α) of internal standards methylparaben/2,6-dimethylpyridine for ODS-AM is different from that of ODS-AL, which is not endcapped. ODS-AL, which is affected by residual silanol, shows relatively great retention of pyridines. ODS-AM 1 1 2 3 2 3 α(4/3)=2.80 α(4/3)=1.46 1. Uracil 2. Pyridine 3. 2,6-Dimethylpyridine 4. Methylparaben 4 4 Utilizes residual silanol groups for separation ODS-AQ ODS-AL The figure shows separation of disinfectants under the conditions described in the Japanese Pharmacopeia. The object is to select the column permitting the elution of benzoic acid, salicylic acid and theophylline, in this order, insuring complete separation of these peaks. With ODS-AQ, separation of salicylic acid and theophylline is incomplete. By contrast, ODS-AL provides excellent separation. Thus, ODS-AL may provide excellent separation when the separation conditions cannot be optimized on other ODS columns. 2,3 3 1 Disinfectants 1 1. 2 COOH 2. COOH OH 4 Benzoic acid Salicylic acid 3. 4. H3C 4 O O H N N N N OH CH 3 Theophylline (I.S.) 0 5 10 15 20 min 0 5 10 15 20 min Phenol Column :YMC-Pack ODS-AL and ODS-AQ (5 µm, 120 Å) 150×4.6 mmI.D. Eluent :100 mM KH2PO4-Na2HPO4 (pH 7.0)/methanol (75/25) Flow rate :0.8 mL/min Temperature:30 ℃ Detection :UV at 270 nm (Conditions described in Japanese Pharmacopoeia 15th ed.) 50 第01章.indd 50 11.1.28 10:41:35 AM Application (G900417B) Phenacyl esters of bile acids from pig gall 2 1. Cholic acid (CA) 2. Hyodeoxycholic acid (HDCA) 3. Chenodeoxycholic acid (CDAC) ( ODS ) Reversed-Phase C18 Columns 3 Column 1 0 10 20 30 40 min :YMC-Pack ODS-AL (5 µm, 120 Å) 250×4.6 mmI.D. Eluent :acetonitrile/water (60/40) Flow rate :1.0 mL/min Temperature:40 ℃ Detection :UV at 240 nm 1 Ordering information Analytical columns Particle size (µm) S-5 Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) Pore size Column size (Å) inner diameter×length(mm) 120 1.0×150 1.0×250 1.5×150 1.5×250 2.0×150 2.0×250 4.6× 75 4.6×100 4.6×150 4.6×250 4.6×300 6.0×100 6.0×150 6.0×250 6.0×300 Semi-preparative columns 10×150 S-5 120 10×250 10×300 20×100 20×150 20×250 Product number AL12S05-1501WT AL12S05-2501WT AL12S05-15P5WT AL12S05-25P5WT AL12S05-1502WT AL12S05-2502WT AL12S05-L546WT AL12S05-1046WT AL12S05-1546WT AL12S05-2546WT AL12S05-3046WT AL12S05-1006WT AL12S05-1506WT AL12S05-2506WT AL12S05-3006WT AL12S05-1510WT AL12S05-2510WT AL12S05-3010WT AL12S05-1020WT AL12S05-1520WT AL12S05-2520WT Particle size (µm) S-5 Pore size Column size (Å) inner diameter×length(mm) 120 1.0×10 1.5×10 2.0×10 4.0×23 Product number AL12S05-0101CC AL12S05-01P5CC AL12S05-0102CC AL12S05-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) Guard columns S-5 120 4.0×10 5.0×10 10×30 20×50 XPCHSMW XPCHW AL12S05-0104WFG AL12S05-0105WFG AL12S05-0310WTG AL12S05-0520WTG See P147 for preparative columns other than those listed above. 51 第01章.indd 51 11.1.28 10:41:36 AM Analytical columns J’sphere ODS-H80, ODS-M80, ODS-L80 ● Useful for method development ● High theoretical plate number ■Pore size:80 Å ■Carbon content:ODS-H80 22% , ODS-M80 14% , ODS-L80 9% ■Usable pH range:ODS-H80 1.0 〜 9.0, ODS-M80・L80 2.0 〜 7.5 Three types of ODS, each with different ligand coverage J’sphere offers a choice from three kinds of ODS made from the same silica gel base, each with different ligand coverage. Differences in ligand coverage considerably affect the hydrophobic retention behavior of solutes, as well as the separation behavior ( ODS ) Reversed-Phase C18 Columns 1 resulting from solute functional groups or tertiary structure. J’sphere is useful for the optimization of separation conditions, since there is almost no need to consider interactions other than hydrophobic and hydrogen-bonding interactions (e.g., ionic or coordinate interaction). Three types of ODS Image of stationary phase ODS-H80 ODS-M80 ODS-L80 CnH 2n+1 (n=0-8) Alkylbenzenes ODS-L80 ODS-M80 Column :J’ sphere 150×4.6 mmI.D. Eluent :acetonitrile/water (75/25) Flow rate :1.0 mL/min Temperature:37 ℃ Detection :UV at 254 nm ODS-H80 0 10 20 30 40 50 min Differences in separation characteristics J’sphere ODS-H80 BRAND-B1 16.5 BRAND-F1 15.9 YMC-Pack ODS-AM 15.2 BRAND-G1 15.2 J’sphere ODS-M80 15.1 BRAND-A2 15.0 BRAND-C3 13.1 6.2 J’sphere ODS-L80 0 10 20 k’(octylbenzene) 0.21 J’sphere ODS-H80 BRAND-F1 0.24 BRAND-G1 0.24 YMC-Pack ODS-AM 0.25 BRAND-C3 0.25 BRAND-B1 0.25 0.27 J’sphere ODS-M80 BRAND-A2 J’sphere ODS-L80 0.20 0.25 α(caffeine/phenol) 29.6 Column :150×4.6 mmI.D. Eluent :acetonitrile/water (75/25) Flow rate :1.0 mL/min Temperature :37 ℃ Detection :UV at 254 nm 30 0.30 0.30 Column :150×4.6 mmI.D. Eluent :acetonitrile/20 mM KH2PO4 (15/85) Flow rate :1.0 mL/min Temperature :37 ℃ Detection :UV at 254 nm ODS columns with different ligand coverage have different hydrophobicity, hydrogen bonding capacity and steric selectivity for tertiary structures. ODS-H80 has exceptionally high hydrophobicity and low hydrogen bonding capacity, ODS-M80 has moderate hydrophobicity and hydrogen bonding capacity and ODS-L80 has low hydrophobicity, comparable to that of C8 and high hydrogen bonding capacity. J’sphere can be used with confidence to identify structural differences of solutes and is useful for improving the efficiency of separation optimization. 0.30 52 第01章.indd 52 11.1.28 10:41:39 AM Ordering information Analytical columns Semi-preparative columns Packing Particle size Pore size Column size material (µm) (Å) inner diameter×length(mm) ODS-H80 ODS-L80 S-4 S-4 80 80 80 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×100 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×150 4.6×250 6.0×150 6.0×250 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 75 2.0×100 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×150 4.6×250 6.0×150 6.0×250 1.0×150 1.0×250 1.5×150 1.5×250 2.0× 35 2.0× 75 2.0×100 2.0×150 2.0×250 3.0×150 3.0×250 4.6× 75 4.6×150 4.6×250 6.0×150 6.0×250 JH08S04-1501WT JH08S04-2501WT JH08S04-15P5WT JH08S04-25P5WT JH08S04-L502WT JH08S04-1002WT JH08S04-1502WT JH08S04-2502WT JH08S04-1503WT JH08S04-2503WT JH08S04-L546WT JH08S04-1546WT JH08S04-2546WT JH08S04-1506WT JH08S04-2506WT JM08S04-1501WT JM08S04-2501WT JM08S04-15P5WT JM08S04-25P5WT JM08S04-L502WT JM08S04-1002WT JM08S04-1502WT JM08S04-2502WT JM08S04-1503WT JM08S04-2503WT JM08S04-L546WT JM08S04-1546WT JM08S04-2546WT JM08S04-1506WT JM08S04-2506WT JL08S04-1501WT JL08S04-2501WT JL08S04-15P5WT JL08S04-25P5WT JL08S04-H502WT JL08S04-L502WT JL08S04-1002WT JL08S04-1502WT JL08S04-2502WT JL08S04-1503WT JL08S04-2503WT JL08S04-L546WT JL08S04-1546WT JL08S04-2546WT JL08S04-1506WT JL08S04-2506WT Packing Particle size Pore size Column size material (µm) (Å) inner diameter×length(mm) ODS-H80 S-4 80 ODS-M80 S-4 80 ODS-L80 S-4 80 10×150 10×250 20×150 20×250 10×150 10×250 20×150 20×250 10×150 10×250 20×150 20×250 Product number JH08S04-1510WT JH08S04-2510WT JH08S04-1520WT JH08S04-2520WT JM08S04-1510WT JM08S04-2510WT JM08S04-1520WT JM08S04-2520WT JL08S04-1510WT JL08S04-2510WT JL08S04-1520WT JL08S04-2520WT Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack) JH08S04-0101CC 1.0×10 ODS-H80 S-4 80 ODS-M80 S-4 80 ODS-L80 S-4 80 1.5×10 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 1.0×10 1.5×10 2.0×10 4.0×23 JH08S04-01P5CC JH08S04-0102CC JH08S04-G304CC JM08S04-0101CC JM08S04-01P5CC JM08S04-0102CC JM08S04-G304CC JL08S04-0101CC JL08S04-01P5CC JL08S04-0102CC JL08S04-G304CC ( ODS ) Reversed-Phase C18 Columns ODS-M80 S-4 Product number 1 A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) XPCHSMW XPCHW YMC guard cartridge pack ODS-H80 S-4 80 4.0×23 JH08S04-G304CCPK ODS-M80 S-4 80 4.0×23 JM08S04-G304CCPK ODS-L80 S-4 80 4.0×23 JL08S04-G304CCPK A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK). Guard columns ODS-H80 S-4 80 ODS-M80 S-4 80 ODS-L80 S-4 80 10×30 20×50 10×30 20×50 10×30 20×50 JH08S04-0310WTG JH08S04-0520WTG JM08S04-0310WTG JM08S04-0520WTG JL08S04-0310WTG JL08S04-0520WTG 53 第01章.indd 53 11.1.28 10:41:40 AM Analytical columns YMC-Pack PolymerC18 ● Utilizes polymer base which is not affected by silanol ● Different separation characteristics from silica-based ODS ● Excellent pH stability ■Usable pH range:2.0 〜 13.0 P o l y m e r t y p e C18 PolymerC18 is a C18 column made from methacrylate polymer. It has excellent pH stability and it is useful for separation of basic compounds because there are no silanol or metal impurities to cause secondary interaction. ( ODS ) Reversed-Phase C18 Columns 1 Since π electrons of the carbonyl group or the hydroxyl group on the surface of the base material show particular interactions with solutes, PolymerC18 can show different separation characteristics from silica-based ODS. Can be used with alkali mobile phase 1 1. Barbital 2. Phenobarbital 3. Pentobarbital 4. Hexobarbital 5. Secobarbital 3 4 2 5 0 10 20 min Column :YMC-Pack PolymerC18 150×4.6 mmI.D. Eluent :50 mM Na2HPO4-Na3PO4(pH 11.0)/methanol (75/25) Flow rate :0.5 mL/min Temperature:30 ℃ Detection :UV at 254 nm Excellent resistance to acid and alkali Alkali resistance Acid resistance 1 1 1 1 〈Prior to eluent flow〉 〈After 6 hours〉 〈Prior to eluent flow〉 〈After 100 hours〉 2 2 3 0 10 20 min 0 10 2 3 20 min 〈Flow conditions〉 Column :YMC-Pack PolymerC18 150×4.6 mmI.D. Eluent :0.1N NaOH (pH 13)/acetonitrile (80/20) Flow rate :0.5 mL/min Temperature :ambient Time :6 hours 〈Analytical conditions〉 Eluent :acetonitrile/water (60/40) Flow rate :0.5 mL/min Temperature :30 ℃ Detection :UV at 270 nm Sample :1.Uracil 2.Methyl benzoate 3.Butyl benzoate 0 3 10 2 20 min 0 10 3 20 min 〈Flow conditions〉 Column :YMC-Pack PolymerC18 150×4.6 mmI.D. Eluent :100 mM H3PO4 (pH 1.7) Flow rate :0.5 mL/min Temperature :ambient Time :100 hours 〈Analytical conditions〉 Eluent :acetonitrile/water (60/40) Flow rate :0.5 mL/min Temperature :30 ℃ Detection :UV at 270 nm Sample :1.Uracil 2.Methyl benzoate 3.Butyl benzoate 54 第01章.indd 54 11.1.28 10:41:43 AM Different separation characteristics from silica-based ODS (aromatic compounds) Polyphenol YMC-Pack ODS-A YMC-Pack PolymerC18 2 1. 3 HO 2. OH OH OH OH Phloroglucinol 2 3. OH Resorcinol Phenol 1 3 0 10 20 30 min 0 Column :YMC-Pack PolymerC18 and YMC-Pack ODS-A 150×6.0 mmI.D. Eluent :methanol/water (40/60) Flow rate :1.0 mL/min Temperature:30 ℃ Detection :UV at 280 nm 5 10 min Different separation characteristics from silica-based ODS (unsaturated compounds) YMC-Pack PolymerC18 Triterpene saponins YMC-Pack ODS-AM 1. 2. 19 ( ODS ) Reversed-Phase C18 Columns 1 1 18 1,2 1 COO-β-D-Glc HO COO-β-D-Glc HO HO HO CH2 OH CH2 OH 2 0.00 7.50 15.00 22.50 min 0.00 7.50 15.00 22.50 min Column :YMC-Pack PolymerC18 and YMC-Pack ODS-AM 150×4.6 mmI.D. Eluent :methanol/water (45/55) for PolymerC18 methanol/water (50/50) for ODS-AM Flow rate :0.8 mL/min Temperature:50 ℃ Detection :UV at 210 nm Ordering information Analytical columns Guard cartridge columns (with inner diameter 2.0 mm: 2/pack, 4.0 mm: 3/pack) Particle size (µm) Column size inner diameter×length(mm) Product number S-6 2.0× 75 2.0×150 4.6×150 4.6×250 6.0×150 6.0×250 4.6×250 4.6×300 6.0×250 6.0×300 PC99S06-L502WT PC99S06-1502WT PC99S06-1546WT PC99S06-2546WT PC99S06-1506WT PC99S06-2506WT PC99S10-2546WT PC99S10-3046WT PC99S10-2506WT PC99S10-3006WT S-10 Semi-preparative columns S-6 S-10 10×250 PC99S06-2510WT 10×250 20×300 PC99S10-2510WT PC99S10-3020WT Particle size (µm) Column size inner diameter×length(mm) Product number S-6 2.0×10 4.0×23 PC99S06-0102CC PC99S06-G304CC A guard cartridge holder will need to be purchased separately before using this product for the first time. Cartridge holder Semi-micro guard cartridge holder (inner diameter: 1.0, 1.5, or 2.0 mm) Cartridge holder (inner diameter: 4.0 mm) Guard columns S-6 S-10 4.0×10 5.0×10 10×30 4.0×10 5.0×10 10×30 20×50 XPCHSMW XPCHW PC99S06-0104WFG PC99S06-0105WFG PC99S06-0310WTG PC99S10-0104WFG PC99S10-0105WFG PC99S10-0310WTG PC99S10-0520WTG 55 第01章.indd 55 11.1.28 10:41:47 AM 第01章.indd 56 11.1.28 10:41:47 AM
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