Transfer Scalable particles: EASY UHPLC HPLC. Flexible ph = 1 12 Temp. up to 70 C 100% aqueous conditions. Universal. Introduction.

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1 YMC-Triart YMC-Triart 3 YMC-Triart Contents Transfer Scalable particles: EASY UHPLC HPLC Universal YMC-Triart for acidic, basic and neutral analytes Flexible ph = Temp. up to 70 C 00% aqueous conditions Phases Overview... 4 First Choice Column for Method Development... 5 Particle Technology... 6 Specifications... 6 ph & Temperature Stability... 7 Method Transfer Between HPLC UHPLC UHPLC YMC-Triart C8 ExRS YMC-Triart Phenyl YMC-Triart PFP LC/MS Pharmaceuticals Food Peptides YMC-Triart AQ / YMC-Triart C HILIC QC Data Ordering Information Introduction Chromatographers always seek to push the limits of HPLC columns to greater extremes to allow them to perform dayto-day with ever-changing ph, buffers and temperature ranges. The column for the laboratory of today must be suitable for harsh ph conditions in combination with high temperature ranges without sacrificing selectivity. In addition narrow, symmetrical peak shapes are necessary in order to cope with rapid analysis of demanding samples. This has required manufacturers to seek more innovative ways to produce suitable stationary phases. In order to meet these goals, YMC has developed a new particle technology. This is based on a particle produced via a tightly controlled granulation technology which has been adapted from micro-reactor technology. The revolutionary production technique provides a silica-organic hybrid stationary phase, which provides an outstandingly narrow pore size and particle size distribution. This in turn, results in low back pressures and high loadability.

2 4 YMC-Triart Phases Overview YMC-Triart 5 YMC-Triart YMC-Triart C8 YMC-Triart C8 ExRS YMC-Triart C8 First choice column for method development Hydrophilic ODS Hydrosphere C8 Si C8 versatile applications first choice for method development ph-stable - Si C8 ExRS extended ph and stability hydrophobic substances positional isomers Si C8 alternative to C8 short retention time Hydrogen bonding capacity a (caffeine/benzene) Pro C8 Triart C8 Pro C8 Triart C8 Symmetry C8 ZORBAX Extend-C8 Standard ODS Pro C8 RS Hydrophobic ODS 00% aqueous eluents ph-stable - ph-stable - Triart C8 ExRS Hydrophobicity k (amylbenzene) 6.00 YMC-Triart Phenyl YMC-Triart PFP YMC-Triart Diol-HILIC Si Phenyl F F F F F HO Si PFP Si HILIC aromatic compounds (p-electron acceptor) conjugated systems 00% aqueous eluents aromatic compounds (p-electron donor) cis-trans isomer polar halogenated compounds 00% aqueous eluents good alternative for very polar compounds 00% aqueous eluents Conventional hybrid silica-based ODS columns tend to be less hydrophobic than silica-based columns. YMC-Triart C8 has a higher carbon load, giving it a hydrophobicity comparable to that of standard ODS columns, thereby making it a versatile first-choice column for method development. On the other hand, YMC-Triart C8 ExRS has been designed to provide contrastingly different separation characteristics! Transfer Scalable particles: EASY UHPLC HPLC Universal YMC-Triart for acidic, basic and neutral analytes Flexible ph = Temp. up to 70 C 00% aqueous conditions

3 6 YMC-Triart YMC-Triart Particle technology YMC-Triart is a versatile material prepared by using tightly controlled particle formation technology which has been adapted from micro-reactor technology. This recently developed production process results in exceptionally narrow particle and pore size distributions. With YMC-Triart, challenging ph and high temperature conditions are no longer a limitation to the day-to-day work in laboratories. Most importantly, due to its unique particle composition, a balanced hydrophobicity and silanol activity are achieved which makes YMC-Triart a First Choice column in method development. Versatile wide ph stability Phosphate buffer (ph.5, 40 C)* YMC-Triart C8 YMC-Triart C8 ExRS Triethylamine (ph.5, 40 C)* YMC-Triart 7 ph & Temperature YMC-Triart C8 YMC-Triart C8 YMC-Triart C8 YMC-Triart C8 ExRS YMC-Triart C8 YMC-Triart Phenyl YMC-Triart PFP YMC-Triart Diol-HILIC Base organic/inorganic silica Stationary phase C8 C8 C8 Phenyl Pentafluorophenyl USP class L L L7 L L43 L0 Particle size.9, 3 and 5 µm Pore size nm 8 nm nm nm nm nm Diol Column: 5 µm, 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: 50 mm K HPO 4 -K 3 PO 4 (ph.5) / methanol (90/0) Flow rate:.0 ml/min Sample: benzyl alcohol Column: 5 µm, 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: 50 mm triethylamine (ph.5) / methanol (90/0) Flow rate:.0 ml/min Sample: benzyl alcohol Specific surface 360 m /g 430 m /g 360 m /g 360 m /g 360 m /g 360 m /g Stability at high temperature Carbon content 0% 5% 7% 7% 5% ph 6.9, 70 C* 0% ph, 70 C* Bonding trifunctional Endcapping multi-stage multi-stage multi-stage multi-stage none none ph range ~ ~ ~ ~ 0 ~ 8 ~ 0 Temp. range ph -7: 70 C, ph 7-: 50 C ph -7: 70 C, ph 7-: 50 C ph -7: 70 C, ph 7-: 50 C 50 C 50 C 50 C Rate on N [%] 00% 80% 60% 40% 0% Silica based C8 YMC-Triart C8 Silica based C8 alkaline-resistant type 00% aqueous eluents 0% Time [h] Column: 5 µm, 50 x.0 mm ID Part No.: TAS05-050WT Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) / acetonitrile (90/0) Flow rate: 0. ml/min Temperature: 70 C Sample: phenol Column: 5 µm, 50 x.0 mm ID Part No.: TAS05-050WT Eluent: acetonitrile / water (60/40) Flow rate: 0. ml/min Temperature: 37 C Sample: butyl benzoate YMC-Triart phases show great chemical stability due to the newly developed hybrid-silica. Even under high ph or high temperature conditions, the lifetime of YMC-Triart phases is more than 0x greater than conventional reversed phase columns.

4 8 YMC-Triart Transfer HPLC YMC-Triart C8 UHPLC Secure your method transfer! Differences in selectivity, retention time, and also peak shapes between different particle sizes of commercially available C8 phases in the same brand (or an alternative as recommended by its manufacture) have been observed. 3 µm TF().43.9 µm TF().47 YMC has addressed this issue of method transfer. YMC-Triart columns show identical selectivity and excellent peak shapes for basic compounds for all 3.0 µm to.9 µm particle sizes. It allows predictable scale up from UHPLC to conventional HPLC and even to semi-preparative LC, and vice versa. Case Studies** X-Bridge BEH C8 and Acquity UPLC BEH C8.5 µm TF().4.7 µm TF().35 Evaluation of method transfer performance!* Hydrogen bonding capacity [a (Caffeine/Benzene)] Hypersil GOLD 5 µm Hypersil GOLD.9 µm Eluent: methanol/water (80/0) [Amylbenzene] methanol/water (30/70) [Caffeine, Benzene] Kinetex C8.6 µm Kinetex C8.7 µm ACQUITY UPLC BEH C8.7 µm XBridge C8 5 µm L-column ODS.0 µm Hydrophobicity [k (Amylbenzene)] Transfer HPLC YMC-Triart C8 3 µm.9 µm L-column ODS 5 µm With the introduction of UHPLC, sub--µm particles became necessary. Therefore smaller particles have been added to existing column lines. Consequently, sub--µm particles may exhibit differences in chromatographic performance. By introducing YMC-Triart, YMC provides matching chromatographic behaviour for all particles sizes! YMC-Triart 9 UHPLC Method transfer between HPLC UHPLC* Kinetex TM C8.6 µm TF() µm TF() 4.94 YMC-Triart C8 50 x.0 mm ID, 5 µm TAS05-50WT 4.5 MPa (650 psi) 5% B (0-3 min) 5-30% B (3-0 min) 30-45% B (0-55 min) 0. ml/min Eluent: A) water/ formic acid (00/0.) B) acetonitrile/ formic acid (00/0.) Detection: UV at 40 nm Injection: µl (5 µg/ml) 55 min Kinetex TM C8 columns show significant peak tailing and have limited scalability due to lack of larger particle sizes. Column: 50 x.0 mm ID or. mm ID Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0. ml/min Detection: UV at 35 nm. Chlorpheniramine (basic). Dextromethorphan (basic) 3. Propyl paraben (internal standard). Oxine-copper (Copper 8-quinolinolate). Asulam 3. Thiram 4. Triclopyr 5. Mecoprop 6. Flazasulfuron 7. Siduron 8. Halosulfuronmethyl 9. Azoxystrobin YMC-Triart C8 50 x.0 mm ID,.9 µm TASP9-050PT 37 MPa (5400 psi) 5% B (0-0.3 min) 5-30% B ( min) 30-45% B ( min) 0.6 ml/min 6 min When transferring the 55 min HPLC method to UHPLC scale, the resolution remains the same although the separation time is reduced to only 6 min. ** These observations might not be representative for all applications.

5 0 YMC-Triart UHPLC YMC-Triart UHPLC & MS Higher resolution and good loadability* YMC-Triart C8 3 TF()=.47.9 µm Determination of Artificial Sweeteners with LC-MS/MS Ascentis Express C8 3.7 µm TF()=5.05 Acesulfam-K Saccharin Cyclamat-Na Non-biological markers of wastewater entries in ground and surface water Kinetex C8.6 µm,3 TF()= µm,3 TF()=4.94 Column: 50 x.0 or. mm ID Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9)/ acetonitrile (65/35) Flow rate: 0. ml/min Detection: UV at 35 nm YMC-Triart C8 shows superior resolution to the other three phases thereby allowing considerable higher loadability. Lower HETP means higher resolution!* 0,05.9 µm YMC-Triart C8 Extracted Ion Chromatogram (XIC) of Acesulfam-K, 0. µg/l Extracted Ion Chromatogram (XIC) of Saccharin, 0. µg/l HETP (mm/plates) 0,00 0,005.7 µm Ascentis Express (Core-shell type).6 µm Kinetex C8 (Core-shell type).7 µm Kinetex C8 (Core-shell type) 0, Linear velocity (mm/sec) Column: 50 x.0 or. mm ID Eluent: acetonitrile / water (60/40) Sample: Butyl benzoate YMC-Triart C8 always shows the lowest HETP compared to the three Core-Shell products over the range of linear velocity applied. Extracted Ion Chromatogram (XIC) of Cyclamat-Na, 0. µg/l Column: YMC-Triart C8, nm,.9 µm, 00 x 3.0 mm ID Part No.: TASP9-003PT LC-System: Agilent 00 HPLC system and CTC Analytics HTC-Pal Autosampler MS/MS System: Applied Biosystems MDS Sciex API 4000, ESI negative Temperature: 35 C Flow rate: 0.3 ml/min Injection: 40 µl, direct injection Eluent: A: H O (containing 0 mmol NH 4 formate) B: Me (containing 0 mmol NH 4 formate) Gradient: Time % B 75 by courtesy of: Thomas Class, Sandro Jooß PTRL Europe, Helmoholtzstraße, Science Park I, D-8908 Ulm

6 YMC-Triart UHPLC YMC-Triart 3 UHPLC Angiotensin I, II and III* Insulin* Tetracycline antibiotics* Betablockers*. Angiotensin I. Angiotensin II 3. Angiotensin III. Bovine insulin. Human insulin 3. Porcine insulin 3. Oxytetracycline. Tetracycline 3. Chlortetracycline 4. Atenolol. Pindolol 3. Nadolol 4. Metoprolol 5. Propranolol Column: YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID Part No.: TASP9-050PT Eluent: 0 mm KH PO 4 + K HPO 4 (ph 7.9) / acetonitrile (/78) Flow rate: 0.7 ml/min Detection: UV at 0 nm Pressure: 70 bar Injection: 0.5 µl Macrolide antibiotics*. Erythromycin. Spiramycin Column: Part No.: Eluent: Gradient: Flow rate: Detection: Pressure: Injection: 0.5 µl Temperature: 30 C Sulpha drugs* YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID TASP9-050PT A) H O + 0.% TFA B) acetonitrile + 0.% TFA 30% B (0 min); 30-3% B (0-5 min); 3% B (55 min) 0.6 ml/min UV at 0 nm 6 bar. Uracil. Sulphathiazole 3. Sulphamerazin 4. Sulphamethaxole 4 Column: YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID Part No.: TASP9-050PT Eluent: 5 mm CH 3 COONH 4 / acetonitrile (87/3) Flow rate: 0.65 ml/min Detection: UV at 80 nm Pressure: 66 bar Injection: µl Fast LC for conventional HPLC* YMC-Triart C8, 5 µm, 50 x.0 mm ID Column: YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID Part No.: TASP9-050PT Eluent: A) 0 mm CH 3 COONH 4 + ammonia (ph 9.0) B) acetonitrile Gradient: 5% B (.0 min); 75% B (-6 min) Flow rate: 0.35 ml/min Pressure: 450 bar Injection: µl 0% B, 0. ml/min, 5.7 MPa 5% B, 0. ml/min, 5.6 MPa 3 Insufficient separation optimisation Baseline separation, but long analysis time Down scaling YMC-Triart C8, 3 µm, 50 x.0 mm ID Column: YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID Part No.: TASP9-050PT Eluent: A) 0 mm K HPO mm KH PO 4 (ph 7.9) B) acetonitrile Gradient: 60% B (0.5 min); 60-70% B ( min); 70% B (3.5 min) Flow rate: 0.45 ml/min Detection: UV at 0 nm Pressure: 50 bar Injection: µl Temperature: 50 C Column: YMC-Trairt C8, nm,.9 µm, 50 x.0 mm ID Part No.: TASP9-050PT Eluent: H O + formic acid (ph.5) / acetonitrile (75/5) Flow rate: 0.75 ml/min Detection: UV at 80 nm Pressure: 740 bar Injection: 0.5 µl Temperature: 50 C Eluent: A) water / HFBA* (00/0.) B) acetonitrile / HFBA* (00/0.) Temperature: 37 C Detection: UV at 90 nm Injection: µl (0. mg/ml) *heptafluorobutyric acid 5% B, 0.4 ml/min,. MPa Fast baseline separation

7 4 YMC-Triart UHPLC YMC-Triart 5 UHPLC Antidepressants* Stevia leaves* Nasal spray*. Nortriptyline. Toluol 3. Imipramine 4. Amitriptyline 3 4. Stevioside hydrate. Rebaudioside A. Maleic acid sodium salt. Tetrahydrozoline HCl 3. Chlorpheniramine maleate 4. Benzethonium chloride Column: YMC-Triart Phenyl,.9 µm (00 x.0 mm ID) Part No.: TPHSP9-00PT Eluent: methanol / 5 mm KH PO 4 (ph 6.0) (65/35) Flow rate: 0.4 ml/min Injection: µl Temperature: 5 C Resorcinol* Column: YMC-Triart Diol-HILIC,.9 µm (00 x 3.0 mm ID) Part No.: TDHSP9-003PT Eluent: acetonitrile / water (85/5) Flow rate: ml/min Detection: UV at 00 nm Injection: µl Temperature: 30 C Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID) Part No.: TASP9-050PT Eluent: A) water % TFA / B) methanol (50/50) Gradient: min A B Flow rate: 0.6 ml/min Detection: UV at 60 nm Injection: 0. µl. Resorcinol. Pyrocatechol 3. -Methylresorcinol 4. 4-Methylcatechol 5.,5-Methylresorcinol 6. 4-Nitrocatechol Eye drop formulation*. Maleic acid sodium salt. Pyridoxine 3. Neostigmine methylsulfate 4. Chlorpheniramine maleate 5. Naphazoline HCl 6. Glycyrrhizin acid ammonium salt Column: YMC-Triart PFP,.9 µm (00 x.0 mm ID ) Part No.: TPFSP9-00PT Eluent: water + 0.% formic acid / acetonitrile + 0.% formic acid (85/5) Flow rate: 0.8 ml/min Detection: UV at 70 nm Injection: 0.5 µl Temperature: 5 C Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID) Part No.: TASP9-050PT Eluent: A water % TFA B) acetonitrile Flow rate: 0.6 ml/min Detection: UV at 65 nm Injection: 0.5 µl Gradient: min A B

8 6 YMC-Triart UHPLC YMC-Triart 7 UHPLC Guaiacol and impurities* 7 Analgesics*. Pyrocatechol. Phenol 3. Guaiacol 4.,-Dimethoxyl benzene 5. Methyl benzoate. Acetaminophene. -Acetamidophenol 3. Caffeine 4. Acetanilide 5. Acetylsalicylic Acid 6. Phenacetine 7. Salicylic Acid Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID ) Part No.: TASP9-050PT Eluent: water / acetonitrile (50/50) Flow rate: 0.7 ml/min Injection: 0.5 µl Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID ) Part No.: TASP9-050PT Eluent: water + formic acid (ph.5) / acetonitrile (50/50) Flow rate: 0.8 ml/min Detection: UV at 40 nm Injection: µl Paracetamol* Hydroxychloroquine and chloroquine*. 4-Aminophenol. Acetaminophenone 3. -Acetamidophenol 4. 4-Acetamidoacetophenone 5. Acetanilide 6. 4-Nitrophenol 7. 4-Chloroacetanilide 8. -Hydroxyacetophenone Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID ) Part No.: TASP9-050PT Eluent: A) water + formic acid (ph.5) / B) acetonitrile Flow rate: 0.7 ml/min Injection: 0.5 µl Gradient: min A B Column: YMC-Triart C8,.9 µm, nm (50 x.0 mm ID ) Part No.: TASP9-050PT Eluent: 0 mm HCO-HCOONH 4 (ph 4.3) / acetonitrile (90/0) Flow rate:.0 ml/min Injection: µl (0 µg/ml) Temperature: 5 C

9 8 YMC-Triart UHPLC YMC-Triart 9 UHPLC Oligonucleotides d(t) -0 method transfer from HPLC to UHPLC* Duloxetine and its degradation products Column: Part No. Flow rate: Gradient: YMC-Triart C8 (5 µm, nm), 50 x.0 mm ID TAS05-50WT 0. ml/min 30-50% B (0-84 min) Column: YMC-Triart C8 (5 µm, nm), 50 x 3.0 mm ID Part No. TAS05-503WT Flow rate: 0.45 ml/min Gradient: 0-90% B (0-36 min) Injection: 6 µl 35 min Column: Part No. Flow rate: Gradient: 80 min YMC-Triart C8 (.9 µm, nm), 50 x.0 mm ID TASP9-050PT 0.8 ml/min 35-50% B (0-7 min) x faster Column: YMC-Triart C8 (.9 µm, nm), 50 x.0 mm ID Part No. TASP9-050PT Flow rate: 0.8 ml/min Gradient: 0-90% B (0-3 min) Injection: µl 3 min x faster 7 min Eluent: A) 0 mm di-n-butylamine-acetic acid (ph 6.0) B) methanol Detection: UV at 69 nm Injection: µl (5 nmol/ml) Temperature: 37 C Eluent: A) 0 mm CH3COONH4 (ph 6.0) B) acetonitrile Detection: UV at 30 nm Temperature: 30 C Sample: Oxidative degradation products of duloxetine hydrochloride* * Sample preparation was performed as described by Veera Reddy. Arava et al. Der Pharma Chemica, 0 4 (4):

10 30 YMC-Triart YMC-Triart C8 ExRS YMC-Triart 3 YMC-Triart C8 ExRS Structural analogs* High hydrophobicity & high steric recognition ability* Basic Compound Coordination Compound Neutral Compounds Polar p-p interaction. Amitriptyline. 8-Quinolinol 3. Testosterone 4. Naphthalene Acidic Compound Hydrophobic 5. Ibuprofen 6. n-propylbenzene 7. n-butylbenzene Steric Cognitive Ability 8. o-terphenyl (steric) 9. Triphenylene (planar) Column: YMC-Triart C8 ExRS, 5 µm, 8 nm (50 x 3.0 mm ID ) Part No.: TAR08S05-503PTH Eluent: THF / acetonitrile (0/90) Flow rate: 0.45 ml/min Detection: UV at 65 nm Injection: 4.5 µl (0 µg/ml) Temperature: 30 C Column: YMC-Triart C8 ExRS, 5 µm, 8 nm (50 x 3.0 mm ID ) Part No.: TAR08S05-503PTH Eluent: 0 mm HCO-HCOONH 4 (ph 4.3) / acetonitrile (90/0) Flow rate:.0 ml/min Injection: µl (0 µg/ml) Temperature: 5 C

11 3 YMC-Triart YMC-Triart Phenyl YMC-Triart 33 YMC-Triart Phenyl Fexofenadine hydrochloride (Japanese Pharmacopoeia)* A) Standard solution * (0.00 mg/ml fexofenadine hydrochloride) Theoretical plate number (Fexofenadine) Tailing factor (Fexofenadine) System suitability requirement result > <.0.00 Fexofenadine hydrochloride (US Pharmacopoeia)* A) Assay preparation * (assay), Reference solution * (related compounds) (0.06 mg/ml fexofenadine hydrochloride) System suitability Tailing factor (Fexofenadine) requirement (assay) result <.0.00 B) Sample solution * ( mg/ml fexofenadine hydrochloride) B) Test solution * (related compounds) ( mg/ml fexofenadine hydrochloride) Column: YMC-Triart Phenyl, 5 µm, nm (50 x 4.6 mm ID) Part No.: TPHS05-546WT Eluent: acetonitrile / buffer * / triethylamine (350/650/3) * Dissolve 7.5 g of NaH PO 4 -H O and 0.96 g of NaClO 4 -H O in 000 ml water, adjust ph.0 with H 3 PO 4 Flow rate:.0 ml/min (adjust the flow rate so that the retention time of fexofenadine is about 9 min) Detection: UV at 0 nm Injection: 0 µl Temperature: 5 C (The Japanese Pharmacopoeia 6th; related substances) Column: YMC-Triart Phenyl, 5 µm, nm (50 x 4.6 mm ID) Part No.: TPHS05-546WT Eluent: acetonitrile / buffer * / triethylamine (350/650/3) * Dissolve 7.5 g of NaH PO 4 -H O and 0.96 g of NaClO 4 -H O in 000 ml water, adjust ph.0 with H 3 PO 4 Flow rate:.5 ml/min Detection: UV at 0 nm Injection: 0 µl Temperature: 5 C (The United States Pharmacopoeia 36th; assay, related compounds) * All standard and sample solutions were prepared from fexofenadine hydrochloride supplied as a reagent for laboratory use. * All standard and sample solutions were prepared from fexofenadine hydrochloride supplied as a reagent for laboratory use. Fexofenadine hydrochloride (Japanese Pharmacopoeia)* Standard solution * (0.06 mg/ml fexofenadine hydrochloride) Fexofenadine hydrochloride (US Pharmacopoeia)* A) Standard solution * (0.05 mg/ml fexofenadine hydrochloride) Tailing factor (Fexofenadine) System suitability requirement (assay) result < Theoretical plate number (Fexofenadine) Tailing factor (Fexofenadine) System suitability requirement result > < B) Sample solution * (0.08 mg/ml fexofenadine hydrochloride) Column: YMC-Triart Phenyl, 5 µm, nm (50 x 4.6 mm ID) Part No.: TPHS05-546WT Eluent: acetonitrile / buffer * / triethylamine (350/650/3) * Dissolve 7.5 g of NaH PO 4 -H O and 0.96 g of NaClO 4 -H O in 000 ml water, adjust ph.0 with H 3 PO 4 Flow rate:.0 ml/min (adjust the flow rate so that the retention time of fexofenadine is about 9 min) Detection: UV at 0 nm Injection: 0 µl Temperature: 5 C (The Japanese Pharmacopoeia 6th; assay) * Standard solutions was prepared from fexofenadine hydrochloride supplied as a reagent for laboratory use. Column: YMC-Triart Phenyl, 5 µm, nm (50 x 4.6 mm ID) Part No.: TPHS05-546WT Eluent: acetonitrile / buffer *3 (9/6) *3 Add 5 ml of acetonitrile/triethylamine (/) to 000 ml of acetic acid/water (7/9983), adjust ph 5.5 with H 3 PO 4 Flow rate:.5 ml/min Detection: UV at 0 nm Injection: 0 µl Temperature: 35 C (The United States Pharmacopoeia 36th; assay) * Standard solution was prepared from fexofenadine hydrochloride supplied as a reagent for laboratory use. * Sample solution was prepared from fexofenadine hydrochloride tablets.

12 34 YMC-Triart YMC-Triart Phenyl YMC-Triart 35 YMC-Triart PFP Analysis of coal tar dye (Brilliant blue FCF) and its impurities* Enlarged chromatogram Catecholamines, serotonin, and their precursors and metabolites* Brilliant blue FCF A-E: Impurities in Brilliant blue FCF reagent Column: YMC-Triart Phenyl, 5 µm, nm (50 x 3.0 mm ID) Part No.: TPHS05-503WT Eluent: water + 0.% phosphoric acid / methanol (55/45) Flow rate: 0.45 ml/min Detection: UV at 630 nm Injection:.0 µl Column: YMC-Triart PFP, 3 µm, nm (50 x 3.0 mm ID) Part No.: TPFS03-503WT Eluent: A) 0 mm formic acid B) methanol containing 0 mm formic acid 0-0% B (0-30 min), 0% B (30-35 min) Flow rate: 0.45 ml/min Detection: UV at 80 nm Injection: 4 µl (5 µg/ml) Temperature: 5 C

13 36 YMC-Triart LC/MS YMC-Triart 37 LC/MS LC/MS compatibility* LC/MS analysis of tetracycline antibiotics* YMC-Triart C8 No column Column: 5 µm, 50 x.0 mm ID Part No.: TAS05-050WT Eluent: A) water / formic acid (00/0.) B) acetonitrile / formic acid (00/0.) 5% B (0- min), 5-00% B (-5 min), 00% B (5-0 min), 00-5% B (0-0. min), 5% B (0.-.5 min) Flow rate: 0.4 ml/min Detection: ESI positive, TIC (Mass Range: ) Column bleeding, caused by the fragments of stationary phase, is the main reason for background noise and restrictions on detection limits. No bleed is observed in the test of total ion current (TIC) measured by LC/MS with blank or with YMC-Triart C8. So in terms of the signal/noise ratio (S/N ratio), YMC-Triart C8 can be expect to not only reduce the background noise but to also increase the sensitivity of the analysis. LC/MS analysis of benzodiazepine derivates Peak : 00ng_mL_RB4_0_9.d: EIC ±0.0+All MS Peak : 00ng_mL_RB4_0_9.d: EIC ±0.0+All MS Courtesy of J. Watanabe, Bruker Daltonics K. K. Column: YMC-Triart C8 (5 µm, nm) 50 x.0 mm ID Part No.: TAS05-050WT Eluent: A) 0 mm formic acid B) acetonitrile Gradient: 5-50% B (0-0 min) Flow rate: 0. ml/min Detection: Bruker Daltonics microtof, ESI, positive mode Injection: 5 µl (00 ng/ml) Column: YMC-Triart C8 (5 µm, nm) 50 x.0 mm ID Part No.: TAS05-050WT Eluent: acetonitrile / water / formic acid (5/85/0.) Flow rate: 0.4 ml/min Detection: A) UV at 70 nm B) ESI positive-mode Injection: 0 µl ( µg/ml)

14 38 YMC-Triart LC/MS YMC-Triart 39 Pharmaceuticals Analysis of 360 pesticides in a single run Separation of alkaloids*. Scopolamine. Atropine 3. Cinchonine 4. Quinine 5. Dihydroquinine Column: YMC-Triart C8 (5 µm, nm) 50 x.0 mm ID Part No.: TAS05-050WT Eluent: 0 mm CH 3 CO-CH 3 COONH 4 (ph 4.9) / acetonitrile (80/0) Flow rate: 0. ml/min Detection: UV at 0 nm Injection: µl ( mg/ml) Barbiturates in human serum* A) Standard (0 µg/ml) Column: YMC-Triart C8 (3 µm, 00 x.0 mm ID) Part No.: TAS03-00WT Eluent: A) 5 mm ammonium formate / water B) 5 mm ammonium formate / methanol Flow rate: 0.5 ml/min Temperature: 45 C Injection: 5 µl Gradient: 0 min: 30% B, 0. min: 50% B, 8 min: 00% B, min: 00% B,.0 min: 30% B, 9 min: 30% B Total run time: 30 min Sample: 00 ng/ml pesticide mix in acetonitrile by courtesy of: József László WIREC, WESSLING International Research and Educational Centre Nonprofit Co. (Hungary) B) Spiked human serum Solid-phase extraction method YMC Dispo SPE C8 00 mg/ml Condition ml methanol ml water Load 500 µl spiked human serum solution (each 0 µg) Elute 500 µl methanol/water (85/5) Dilute 500 µl 0 mm ammonium formate buffer (ph 9.5) Column: YMC-Triart C8 (5 µm, nm) 50 x.0 mm ID Part No.: TAS05-050WT Eluent: A) 0 mm HCOONH 4 -NH 3 (ph 9.5) B) methanol Gradient: 0-90% B (0-7 min) Flow rate: 0. ml/min Temperature: 5 C Detection: UV at 40 nm Injection: µl

15 40 YMC-Triart Pharmaceuticals YMC-Triart 4 Pharmaceuticals Erythromycin at elevated ph and temperature*. Optimisation of ph Separation of flavonoids*. Optimisation of temperature (ph 7.9) Column: YMC-Triart C8 (5 µm, nm) Dimension: 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: acetonitrile / 0 mm H 3 PO 4 (30/70) Flow rate: 0.45 ml/min Temperature: 37 C Detection: UV at 80 nm Injection: µl (50 µg/ml) Separation of aromatic carboxylic acids* 3. Stability test: ph 7.9, 70 C Number of injections Column: YMC-Triart C8 (5 µm, nm) Dimension: 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: 0 mm CH 3 CO-CH 3 COONH 4 (ph 4.) / acetonitrile (75/5) Flow rate: 0.45 ml/min Injection: 4 µl (0.0 ~ 0.3 mg/ml) Column: YMC-Triart C8 (3 µm, nm) Dimension: 50 x.0 mm ID Part No.: TAS03-050WT Eluent: 0 mm KH PO 4 -K HPO 4 / acetonitrile / methanol (40/45/5) Flow rate: 0. ml/min Detection: UV at 0 nm

16 4 YMC-Triart Pharmaceuticals YMC-Triart 43 Pharmaceuticals Aciclovir syrup and injection* Basic drugs*. Hydrochlorothiazide. Amlodipine besilate 3. Valsartan 4. Atorvastatin calcium hydrate 5. Candesartan cilexetil Column: YMC-Triart C8 (5 µm, nm) 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: phosphate buffer* / methanol (95/5) *Dissolve.45 g of H 3 PO 4 and 5 ml of mol/l CH 3 CO in water to make 900 ml q adjust ph.5 by mol/l Na q add water to make 000 ml Flow rate:.0 ml/min Temperature: 5 C Injection: 0 µl (0.05 mg/ml, 0.03 mg/ml) YMC-Triart C8 Clindamycin hydrochloride* Column: YMC-Triart C8 (3 µm, nm), 50 x.0 mm ID Part No.: TOS03-050WT Eluent: A) water / formic acid (00/0.) B) acetonitrile / formic acid (00/0.) 0-90% B (0-5 min), 90% B (5-7 min) Flow rate: 0.4 ml/min Temperature: 30 C Injection: µl (0-0 µg/ml) Sequential analysis of Carvedilol* Column: YMC-Triart C8 (5 µm, nm) 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: 50 mm KH PO 4 (ph 7.5 adjusted by 8 M K) / acetonitrile (55/45) Flow rate:.0 ml/min Temperature: 5 C Detection: UV at 0 nm Injection: 0 µl Analysis of amlodipine besilate* Column: YMC-Triart C8 (5 µm, 50 x.0 mm ID) Part No.: TOS05-50WT Eluent: phosphate buffer (ph.0)* / acetonitrile (65/35) *Dissolve.7 g of KH PO 4 in 900 ml water, adjust ph.0 with H 3 PO 4 and add water to make 000 ml Flow rate: 0.8 ml/min (adjust the flow rate so that the retention time of carvedilol is about 4 min) Temperature: 55 C Detection: UV at 40 nm YMC-Triart C8 No change in retention time is observed even under a high ph and at an elevated temperature. Column: YMC-Triart C8 (5 µm, nm) 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: 0 mm CH 3 CO-CH 3 COONH 4 (ph 4.) / acetonitrile (75/5) Flow rate: 0.45 ml/min Injection: 4 µl (0.0 ~ 0.3 mg/ml)

17 44 YMC-Triart Food YMC-Triart 45 Food Soy isoflavones in supplement* A) Standard (0.0 mg/ml) B) Supplement ( g/50 ml). Daidzin. Glycitin 3. Genistin O-Malonyldaidzin O-Malonylglycitin O-Acetyldaidzin O-Acetylglycitin O-Malonylgenistin 9. Daidzein 0. Glycitein. 6 -O-Acetylgenistin. Genistein Sample preparation method Supplement Stirred at r.t. for hr Filtration Injection 50% aqueous ethanol Analysis of anthocyanins and anthocyanidins*. Delphinidin-3-O-galactoside. Delphinidin-3-O-glucoside 3. Cyanidin-3-O-galactoside Anthocyanins: Anthocyanidins: Indicated in black Indicated in red 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 0. Petunidin-3-O-arabinoside. Delphinidin. Peonidin-3-O-glucoside 3. Malvidin-3-O-galactoside 4. Peonidin-3-O-arabinoside 5. Malvidin-3-O-glucoside 6. Malvidin-3-O-arabinoside 7. Cyanidin 8. Petunidin 9. Peonidin 0. Malvidin Column: YMC-Triart C8 (5 µm, nm) Dimension: 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: A) water / formic acid (90/0) B) acetonitrile / methanol / water / formic acid (.5/.5/40/0) Gradient: 0-8% B (0-30 min), 8-70% B (30-40 min), 00% B (40-45 min) Flow rate:.0 ml/min Temperature: 5 C Detection: UV/VIS at 535 nm Sample: commercial bilberry powder (.5 mg/ml) Column: YMC-Triart C8 (3 µm, nm) 50 x.0 mm ID Part No.: TAS03-050WT Eluent: A) acetonitrile / water / HCO (0/90/0.) B) acetonitrile / water / HCO (60/40/0.) Separation of water-soluble vitamins* Gradient: 5-40% B (0- min) Flow rate: 0.4 ml/min Temperature: 5 C Injection: µl Peptides. Thiamine HCl (Vitamin B ). Pyridoxine HCl (Vitamin B 6 ) 3. Nicotinamide 4. Cyanocobalamin (Vitamin B ) 5. L-Ascorbic acid -glucoside 6. L-Ascorbic acid (Vitamin C) 7. Erythorbic acid 8. Riboflavin (Vitamin B ) 9. Nicotinic acid Peptides (MW 556-3,465)*. Oxytocin. Met-Enkephalin 3. Leu-Enkephalin 4. Neurotensin 5. γ-endorphin 6. β-endorphin (MW,007) (MW 574) (MW 556) (MW,673) (MW,859) (MW 3,465) Column: YMC-Triart C8 (5 µm, nm) 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: phospate buffer* / acetonitrile (90/0) * Dissolve.4 g KH PO 4 in 800 ml water q add 6 ml 0% TBA q adjust ph 5. by 0% H 3 PO 4 q add water to make 000 ml Flow rate: 0.8 ml/min Detection: UV at 60 nm Injection: 0 µl (5 µg/ml) Column: YMC-Triart C8 (5 µm, nm) Dimension: 50 x.0 mm ID Part No.: TAS05-50WT Eluent: A) water / TFA (00/0.) B) acetonitrile / TFA (00/0.) 0-45% B (0-5 min) Flow rate: 0. ml/min Temperature: 37 C Detection: UV at 0 nm Injection: ml (0.075 ~ 0.5 mg/ml)

18 46 YMC-Triart YMC-Triart AQ YMC-Triart C8 YMC-Triart 47 YMC-Triart AQ YMC-Triart C8 03 General The use of 00% water eluent has been a challenge in HPLC analysis for decades. Even today, many C8 materials suffer from unacceptable short lifetime, because the C8 chains collapse and this reduces the separation performance drastically. As a pioneer in this field, YMC has offered a product as early as the 80 s, which presents a synonym for stability under aqueous conditions: YMC-Pack ODS-AQ. AQ -type phases are particularly suitable for the separation of polar substances, metabolites, pesticides, degradation products and peptides including protein digests. 000 Hydrosphere C8 A new, ultrapure silica base was introduced whilst adapting the surface chemistry to maintain the AQ -type properties. hydrophilic C8 surface for enhanced polar recognition stable when used with 00% aqueous eluent no need for ion pair reagents addition of µm particle size for Fast-LC (YMC-Pack UltraHT) 5 µm µm µm 3 YMC-Triart "AQ" = YMC-Triart C8 The latest YMC technology platform consists of a hybrid-style substrate with enhanced stability against ph - temp. up to 70 C 00% HO 5 µm 3 µm.9 µm 985 YMC-Pack ODS-AQ hydrophilic C8 balanced surface chemistry polar recognition metabolite recognition Therefore, YMC-Triart provides freedom in method development, robustness and enhanced column lifetimes for reproducible results, day after day, year after year. YMC-Triart is fully scalable within (U)HPLC HPLC with its micron particle sizes in order to facilitate lab-tolab method transfer. 5 µm 3 µm

19 48 YMC-Triart YMC-Triart AQ YMC-Triart C8 YMC-Triart 49 YMC-Triart AQ YMC-Triart C8 Problem with conventional C8 columns Solution with YMC-Triart C8 Aqueous conditions deteriorate column performance* Example Example Reproducible stable performance!* No retention time changes! Before pump stopped tr 5 = tr 5 = 4 5 Before pump stopped 3 4 tr 5 = 4 5 After pump stopped 63.5% of initial retention 8.% of initial retention After pump stopped 98.0% of initial retention min min Column: 5 µm, 50 x 4.6 mm ID XBridge C8 (Example ) and Atlantis T3 (Example ) Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) Flow rate:.0 ml/min Temperature: 37 C Sample:. Cytosine. Uracil 3. Guanine 4. Thymine 5. Adenine Column: YMC-Triart C8, (5 µm, nm) 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) Flow rate:.0 ml/min Temperature: 37 C min Why? Image of C8 surface hydration The columns used for applications involving 00% aqueous buffers provide shorter retention times after the flow was stopped between analyses. Image of C8 surface hydration When Triart C8 columns are used for applications involving 00% aqueous buffers, the retention times are unchanged after the flow was stopped between analyses. This behaviour is caused by poor hydration of the phase. Polar compounds cannot easily distribute between the mobile phase and the stationary phase. This is due to the improved hydration of the phase. Polar compounds can easily distribute between the mobile phase and the stationary phase.

20 50 YMC-Triart YMC-Triart 5 YMC-Triart AQ YMC-Triart C8 YMC-Triart AQ YMC-Triart C8 From the inventors of AQ-columns: YMC-Triart C8 "validated" for AQ-conditions!* 0 HPLC UHPLC 5 µm, 50 x 3.0 mm ID, µm, 00 x 3.0 mm ID, ml/min 4 N(4)=900 8 MPa 0min 0.85 ml/min 4 N(4)= MPa. Oxalic acid. Tartaric acid 3. Glycolic acid 4. Formic acid 5. L-Malic acid 6. Malonic acid 7. Lactic acid 8. Acetic acid 9. Maleic acid 0. Citric acid. Succinic acid. Fumaric acid 3. Acrylic acid 4. Propionic acid Proven reliability* min 500 th 400 th 300 th 00 th 00 th st (%) Retention stability of Adenine Analysis number Adenine 0 Column: YMC-Triart C8 Eluent: 0 mm H 3 PO 4 Flow rate: 0.45 ml/min Temperature: 37 C Detection: 0 nm 3 4min 00% aqueous conditions! Column: YMC-Triart C8, (5 µm, nm) 50 x 3.0 mm ID Part No.: TAS05-530WT Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) Flow rate: 0.45 ml/min Temperature: 37 C Sample:. Cytosine. Uracil 3. Guanine 4. Thymine 5. Adenine No change is found in the separation parameters including retention times, even after 500 injections when using YMC-Triart C8. Stable under harsh conditions: ph - and temperature up to 70 C. Stable retention times with 00% aqueous eluents! Reproducible results day after day, column to column and lab to lab! YMC-Triart C8 Hydrosphere C8 YMC-Pack ODS-AQ , min min. L-Ascorbic acid -glucoside. L-Ascorbic acid 3. D-Isoascorbic acid 4. Nicotinic acid 5. Nicotinamide 6. D-(+)-Pantothenic acid calcium salt 7. Pyridoxal hydrochloride 8. Pyridoxine hydrochloride 9. Folic acid 0. Thiamine hydrochloride. Cyanocobalamin. D-Biotin 3. Riboflavin min Column: 3 µm, 50 x 3.0 mm ID Part No.: TAS05-530WT Eluent: A) 0 mm KH PO 4 -H 3 PO 4 (ph.8) containing 5 mm CH 3 (CH )5SO 3 Na B) 0 mm KH PO 4 -H 3 PO 4 (ph.8) / acetonitrile (80/0) containing 5 mm CH 3 (CH )5SO 3 Na Flow rate: 0.45 ml/min Temperature: 40 C Detection: 0 nm Retention behaviour of water-soluble vitamins on three YMC ODS phases which can be used with 00% aqueous mobile phases is compared. The retention times and peak elution order for folic acid (peak 9), thiamine hydrochloride (peak 0) and cyanocobalamin (peak ) are different for the three phases due to the balance of hydrophobicity and hydrogen bonding capacity differing between the three phases.

21 5 YMC-Triart YMC-Triart 53 HILIC HILIC Great stability and reproducibility at high ph* Polar and hydrophilic compounds* Stability in high ph (ph, 50 C)** Rate on N [%] Triart Diol-HILIC 00% 80% 60% 40% Cytosine 00 hr 50 hr 00 hr mv 00 RP mode YMC-Triart C8 0 mm KH PO 4 -H 3 PO 4 (ph.5) mv 00 HILIC mode YMC-Triart Diol-HILIC 00 mm HCO-HCOONH 4 (ph 3.3) / acetonitrile (0/90) 0% Silica based Diol 0% ** ph < 0 is recommended for regular use Time [h] hr initial min Column: 5 µm, 50 x 4.6 mm ID Part No.: TDHS05-546WT Eluent: acetonitrile / water / NH 3 (90/0/0.) ph.3 Flow rate:.0 ml/min Temperature: 50 C Sample: Cytosine YMC-Triart Diol-HILC offers highly reproducible separations even at high ph and high temperature. The lifetime of YMC- Triart Diol-HILIC is much longer than that of conventional silica-based Diol columns. 0 F045A min 0 B047L min Nucleosides and bases* mau Uracil. Uridine 3. Adenosine 4. Cytosine 5. Cytidine 6. Guanosine Diabetes drugs* mau NH NH H N NH NH H Cl Phenformin hydrochloride NH NH H N NH N CH 3 HCl CH 3 Metformin hydrochloride HO H O O HO HO HO Erythorbic acid (D-Isoascorbic acid) HO H O O HO L-Ascorbic acid Column: 5 µm, 50 x 3.0 mm ID Flow rate: 0.45 ml/min Injection: 4 µl (0.05 mg/ml) YMC-Triart C8 (RP) shows very weak retention and poor resolution of L-ascorbic acid and its stereoisomer (erythorbic acid) even if 00% aqueous mobile phase is used. However, YMC-Triart Diol-HILIC shows strong retention and good resolution of these compounds with mobile phase containing 90% organic solvent. 0 V050A min 0 U059D min Column: YMC-Triart Diol-HILIC (5 µm, nm) 50 x 3.0 mm ID Part No.: TDHS05-503WT Eluent: 00 mm CH 3 COONH 4 / acetonitrile (0/90) Flow rate: 0.45 ml/min Temperature: 30 C Injection: µl (5 ~ 0 µg/ml) Column: YMC-Triart Diol-HILIC (.9 µm, nm) 50 x.0 mm ID Part No.: TDHSP9-050PT Eluent: 00 mm HCO-HCOONH 4 (ph 3.7) / acetonitrile (0/90) Flow rate: 0.8 ml/min Temperature: 5 C Detection: UV at 35 nm Injection: µl (0 µg/ml)

22 54 YMC-Triart QC Data YMC-Triart 55 QC Data YMC-Triart: Improved quality of particles Uniform spherical particles YMC-Triart X-Bridge HILIC Excellent reproducibility of YMC-Triart phases is available on consecutive lots even for the analysis for basic and coordination compounds which normally exhibit tailing and adsorption effects. Batch-to-batch reproducibility Basic compounds* 3. Chlorpheniramine (basic). Dextromethorphan (basic) 3. Propyl p-hydroxybenzoate Coordination compounds*. Hinokitiol (coordination). Methyl benzoate Lot. E Lot. E Lot. D Lot. D Lot. C Lot. C Lot. B Lot. B J090Y min Lot. A J090X min Lot. A by courtesy of YMC Co., Ltd. The uniform spherical particle support is used for all YMC-Triart phases. The particles are produced using micro-reactor technology for the granulation process. This results in reduction of the backpressure and leads to more reproducibility in surface modification. Column: YMC-Triart C8, 5 µm, 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: 0 mm KH PO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.45 ml/min Detection: UV at 35 nm Column: YMC-Triart C8, 5 µm, 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: acetonitrile / 0.% H 3 PO 4 (40/60) Flow rate: 0.45 ml/min Low column back-pressure* YMC-Pack Pro C8, methanol/h O YMC-Pack Pro C8, acetonitrile/h O YMC-Triart C8, methanol/h O YMC-Triart C8, acetonitrile/h O Column: YMC-Triart C8, 5 µm, 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: acetonitrile / water or methanol / water Flow rate:.0 ml/min Temperature: 5 C The revolutionary production technique, adapted from micro-reactor flow technology, produces a silica/organic hybrid stationary phase, with outstanding narrow pore size and particle size distributions which result in low back pressures. YMC-Triart is designed for use under a wide range of conditions. Elution with higher viscosity methanol (compared with acetonitrile), YMC-Triart generates lower pressure (approx 30% lower than with conventional phases). The reproducibility of packed columns is shown below in terms of theoretical plate number (N) and tailing factor (Tf). YMC-Triart packed columns exhibit a very narrow range of variation. Theoretical plate number Theoretical plate number Column number Tailing factor Tailing factor Column: YMC-Triart C8, 5 µm, 50 x 4.6 mm ID Part No.: TAS05-546WT Eluent: acetonitrile / water (40/60) Flow rate: Temperature: Sample:.0 ml/min ambient butyl benzoate

23 56 YMC-Triart QC Data YMC-Triart 57 QC Data Narrow pore distribution Multi-stage endcapping d Vp/ d logd YMC-Triart C8 particle Conventional silica-based particle X-Bridge HILIC After bonding the alkyl chain, there are highly reactive and less reactive silanols on the surface. In traditional bonding processes, these are reacted with a single endcapping-compound in one step. However, the highly reactive silanols can be hydrolysed easily which contributes to the poor stability. The less reactive silanols are hard to endcap which results in poor resolution due to peak tailing. YMC-Triart phases use an innovation in endcapping called multistage endcapping for its surface modification process. By using a number of compounds with different reactivities in successive steps, all silanols can be capped to the maximum extent. Before EC STEP Pore size (nm) This figure shows the pore size distributions of some competitive stationary phases. Comparing the pore size distributions shows a narrower distribution for YMC-Triart which results in sharper peak shapes Hydroxyl group Endcapping (EC) group Improved loadability Influence of injection volume on peak shapes YMC-Triart C8 C8 solvent for sample acetonitrile injection volume.0 µl.5 µl.0 µl 3.0 µl STEP 3 STEP Hybrid-silica based C8. OCH CHCH NHCH(CH 3 ) Propranolol (50 µg/ml). H 3 CO CN CH 3 OCH 3 The chromatographic result of a good endcapping is demonstrated:.5 Peak anomaly at.5 µl.0 Silica-based C8.5 min H 3 CO C (CH ) 3 NCH CH CH(CH 3 ) CH 3 Verapamil (50 µg/ml) OCH 3 Column: 5 µm, 50 x.0 or. mm ID Eluent: A) water / formic acid (00/0.) B) acetonitrile / formic acid (00/0.) 5% B (0-0.5 min), 5-00% B ( min) Flow rate: 0.4 ml/min Detection: UV at 75 nm Endcapping by traditional method Peak tailing for basic compounds YMC-Triart C8 Ingredients in cough/cold medication:. Chlorpheniramine. Dextromethorphan 3. Propyl p-hydroxybenzoate In order to prevent peak errors, there is a limit to the injection volume when a sample is injected in high elution solvents (such as 00% acetonitrile). Compared with traditional columns, more than double the injection volume can be injected into YMC-Triart columns as a result of the extremely narrow particle size distribution. Column: 5 µm, 50 x 3.0 mm ID Part No.: TAS05-503WT Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.45 ml/min Detection: UV at 35 nm

24 58 YMC-Triart QC Data YMC-Triart 59 QC Data Basic compounds* YMC-Triart C8 50 x 3.0 mm ID silica based C8 Inertsil 50 x 4.6 mm ID hybrid-silica based C8 XBridge 50 x 4.6 mm ID Ingredients in a cough/cold medication. Chlorpheniramine. Dextromethorphan 3. Propyl p-hydroxybenzoate excellent peak shape Column: 5 µm, 50 x 3.0 or 50 x 4.6 mm ID Eluent: 0 mm KH PO 4 -K HPO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.45 ml/min for 3.0 mm ID.0 ml/min for 4.6 mm ID Detection: UV at 35 nm The innovative surface modification technology results in excellent peak shapes even for basic compounds that often exhibit peak tailing with conventional silica- and hybrid silica-based reversed phase columns. Comparison of clemastine analysis* 0 mm phosphate buffer (ph 6.7)/organic solvent 0 mm CH3COONH4/organic solvent methanol acetonitrile methanol acetonitrile YMC-Triart C8 C8 50 X x.0 mmi.d. ID Tf 0.97 Tf. Tf.03 Tf.6 Silica based C8 C8 Inertsil Brand ODS-3V C 50 X x. mmi.d. ID F09088H07 F09088I07 F09088F07 F09087G07 Tf. Tf.56 Tf.3 Tf.43 F09077H07 F09077I07 F09077F07 F090803G07 Acidic compounds* YMC-Triart C8 50 x 3.0 mm ID Organic acid. Formic acid. Acetic acid 3. Propionic acid Silica Silica based based C8 C8 Capcell Brand D5 Pak MG-II 50 X x.0 mmi.d. ID Silica based C8 Silica based C8 Sunfire C8 Brand I x X.. mmi.d. mm ID Tf 0.93 Tf.83 Tf.05 Tf. F09077H07 F09077I07 F09074F07 F09074G07 Tf.03 Tf 3.87 Tf.3 Tf.88 F0907H07 F0907I07 F0907F07 F0907G07 hybrid-silica based C8 Gemini NX 50 x 4.6 mm ID Column: 5 µm, 50 x 3.0 or 50 x 4.6 mm ID Eluent: methanol / 0.% H 3 PO 4 (5/95) Flow rate: 0.45 ml/min for 3.0 mm ID.0 ml/min for 4.6 mm ID Temperature: 37 C Detection: UV at 0 nm Hybrid-silica based C8 X-Bridge Tf.4 Tf.88 Tf. 50 x. mm ID F09073H07 F09073I07 F09073F min min min Tf.78 F09073G min YMC-Triart phases are synthesised using methodology adapted from micro-reactor technology. This technique ensures a reduction of impurities that contribute to peak tailing during the analysis of some types acidic compounds. Column: 5 µm, 50 x.0 or 50 x. mm ID Coordinating compounds* YMC-Triart C8 50 x 3.0 mm ID Hinokitiol. Hinokitiol. Methyl benzoate low metal impurities Eluent: A) 0 mm KH PO 4 -K HPO 4 (ph 6.7) or 0 mm CH 3 COONH 4 B) methanol or acetonitrile 5-90% B (0-0 min), 90% B (0-5 min) Flow rate: 0. ml/min Temperature: 5 C Detection: UV at 30 nm Clemastine is a well-known basic compound which readily exhibits peak tailing with conventional ODS columns. YMC- Triart C8 provides sharp separations with many different buffer/solvent compositions. hybrid-silica based C8 XBridge 50 x 4.6 mm ID Column: 5 µm, 50 x 3.0 or 50 x 4.6 mm ID Eluent: acetonitrile / 0.% H 3 PO 4 (40/60) Flow rate: 0.45 ml/min for 3.0 mm ID.0 ml/min for 4.6 mm ID YMC-Triart phases have an extremely low level of metal impurities, much lower than conventional products, ensuring excellent peak shape for coordination compounds.

25 60 YMC-Triart Ordering Information YMC-Triart.9 µm UHPLC columns (max. pressure,000 bar) Phase C8 C8 ExRS C8 Phenyl PFP HILIC Column ID (mm) Column length (mm) Guard cartridges* with 5 mm length 0 30/ (pack of 3).0 TASP9-00PT TASP9-030PT TASP9-050PT TASP9-L50PT TASP9-00PT TASP9-50PT TASP9-E5QCC** 3.0 TASP9-0503PT TASP9-L503PT TASP9-003PT TASP9-503PT TASP9-E503CC.0 TAR08SP9-00PT TAR08SP9-H30PT TAR08SP9-050PT TAR08SP9-L50PT TAR08SP9-00PT TAR08SP9-50PT TAR08SP9-E5QCC** 3.0 TAR08SP9-0503PT TAR08SP9-L503PT TAR08SP9-003PT TAR08SP9-503PT TAR08SP9-E503CC.0 TOSP9-00PT TOSP9-030PT TOSP9-050PT TOSP9-L50PT TOSP9-00PT TOSP9-50PT TOSP9-E5QCC** 3.0 TOSP9-0503PT TOSP9-L503PT TOSP9-003PT TOSP9-503PT TOSP9-E503CC.0 TPHSP9-00PT TPHSP9-030PT TPHSP9-050PT TPHSP9-L50PT TPHSP9-00PT TPHSP9-50PT TPHSP9-E5QCC** 3.0 TPHSP9-0503PT TPHSP9-L503PT TPHSP9-003PT TPHSP9-503PT TPHSP9-E503CC.0 TPFSP9-00PT TPFSP9-030PT TPFSP9-050PT TPFSP9-L50PT TPFSP9-00PT TPFSP9-50PT TPFSP9-E5QCC** 3.0 TPFSP9-0503PT TPFSP9-L503PT TPFSP9-003PT TPFSP9-503PT TPFSP9-E503CC.0 TDHSP9-050PT TDHSP9-L50PT TDHSP9-00PT TDHSP9-E5QCC** 3.0 TDHSP9-0503PT TDHSP9-L503PT TDHSP9-003PT *Guard cartridge holder required, part no. XPCHUHP **Guard cartridge:. mm ID Phase Column ID (mm) Column length (mm) YMC-Triart 6 Ordering Information YMC-Triart 3 µm high pressure rated analytical columns (max. pressure 450 bar) Phase C8 C8 ExRS C8 Phenyl PFP HILIC Column ID (mm) Column length (mm) YMC-Triart 5 µm high pressure rated analytical columns (max. pressure 450 bar) Guard cartridges* with 0 mm length (pack of 5). TAS03-0QPTH TAS03-H3QPTH TAS03-05QPTH TAS03-L5QPTH TAS03-0QPTH TAS03-5QPTH TAS03-0QGC 3.0 TAS PTH TAS03-L503PTH TAS03-003PTH TAS03-503PTH TAS03-003GC 4.6 TAS03-H346PTH TAS PTH TAS03-L546PTH TAS03-046PTH TAS03-546PTH TAS03-546PTH TAS03-004GC. TAR08S03-0QPTH TAR08S03-H3QPTH TAR08S03-05QPTH TAR08S03-L5QPTH TAR08S03-0QPTH TAR08S03-5QPTH TAR08S03-0QGC 3.0 TAR08S PTH TAR08S03-L503PTH TAR08S03-003PTH TAR08S03-503PTH TAR08S03-003GC 4.6 TAR08S03-H346PTH TAR08S PTH TAR08S03-L546PTH TAR08S03-046PTH TAR08S03-546PTH TAR08S03-546PTH TAR08S03-004GC. TOS03-0QPTH TOS03-H3QPTH TOS03-05QPTH TOS03-L5QPTH TOS03-0QPTH TOS03-5QPTH TOS03-0QGC 3.0 TOS PTH TOS03-L503PTH TOS03-003PTH TOS03-503PTH TOS03-003GC 4.6 TOS03-H346PTH TOS PTH TOS03-L546PTH TOS03-046PTH TOS03-546PTH TOS03-546PTH TOS03-004GC. TPHS03-0QPTH TPHS03-H3QPTH TPHS03-05QPTH TPHS03-L5QPTH TPHS03-0QPTH TPHS03-5QPTH TPHS03-0QGC 3.0 TPHS PTH TPHS03-L503PTH TPHS03-003PTH TPHS03-503PTH TPHS03-003GC 4.6 TPHS03-H346PTH TPHS PTH TPHS03-L546PTH TPHS03-046PTH TPHS03-546PTH TPHS03-546PTH TPHS03-004GC. TPFS03-0QPTH TPFS03-H3QPTH TPFS03-05QPTH TPFS03-L5QPTH TPFS03-0QPTH TPFS03-5QPTH TPFS03-0QGC 3.0 TPFS PTH TPFS03-L503PTH TPFS03-003PTH TPFS03-503PTH TPFS03-003GC 4.6 TPFS03-H346PTH TPFS PTH TPFS03-L546PTH TPFS03-046PTH TPFS03-546PTH TPFS03-546PTH TPFS03-004GC. TDHS03-0QPTH TDHS03-H3QPTH TDHS03-05QPTH TDHS03-L5QPTH TDHS03-0QPTH TDHS03-5QPTH TDHS03-0QGC 3.0 TDHS PTH TDHS03-L503PTH TDHS03-003PTH TDHS03-503PTH TDHS03-003GC 4.6 TDHS03-H346PTH TDHS PTH TDHS03-L546PTH TDHS03-046PTH TDHS03-546PTH TDHS03-546PTH TDHS03-004GC *Guard cartridge holder required, part no. XPGCH-Q Guard cartridges* with 0 mm length (pack of 5) For other dimensions please refer to page C8 C8 ExRS C8 Phenyl PFP HILIC. TAS05-0QPTH TAS05-H3QPTH TAS05-05QPTH TAS05-L5QPTH TAS05-0QPTH TAS05-5QPTH TAS05-0QGC 3.0 TAS PTH TAS05-L503PTH TAS05-003PTH TAS05-503PTH TAS05-003GC 4.6 TAS05-H346PTH TAS PTH TAS05-L546PTH TAS05-046PTH TAS05-546PTH TAS05-546PTH TAS05-004GC. TAR08S05-0QPTH TAR08S05-H3QPTH TAR08S05-05QPTH TAR08S05-L5QPTH TAR08S05-0QPTH TAR08S05-5QPTH TAR08S05-0QGC 3.0 TAR08S PTH TAR08S05-L503PTH TAR08S05-003PTH TAR08S05-503PTH TAR08S05-003GC 4.6 TAR08S05-H346PTH TAR08S PTH TAR08S05-L546PTH TAR08S05-046PTH TAR08S05-546PTH TAR08S05-546PTH TAR08S05-004GC. TOS05-0QPTH TOS05-H3QPTH TOS05-05QPTH TOS05-L5QPTH TOS05-0QPTH TOS05-5QPTH TOS05-0QGC 3.0 TOS PTH TOS05-L503PTH TOS05-003PTH TOS05-503PTH TOS05-003GC 4.6 TOS05-H346PTH TOS PTH TOS05-L546PTH TOS05-046PTH TOS05-546PTH TOS05-546PTH TOS05-004GC. TPHS05-0QPTH TPHS05-H3QPTH TPHS05-05QPTH TPHS05-L5QPTH TPHS05-0QPTH TPHS05-5QPTH TPHS05-0QGC 3.0 TPHS PTH TPHS05-L503PTH TPHS05-003PTH TPHS05-503PTH TPHS05-003GC 4.6 TPHS05-H346PTH TPHS PTH TPHS05-L546PTH TPHS05-046PTH TPHS05-546PTH TPHS05-546PTH TPHS05-004GC. TPFS05-0QPTH TPFS05-H3QPTH TPFS05-05QPTH TPFS05-L5QPTH TPFS05-0QPTH TPFS05-5QPTH TPFS05-0QGC 3.0 TPFS PTH TPFS05-L503PTH TPFS05-003PTH TPFS05-503PTH TPFS05-003GC 4.6 TPFS05-H346PTH TPFS PTH TPFS05-L546PTH TPFS05-046PTH TPFS05-546PTH TPFS05-546PTH TPFS05-004GC. TDHS05-0QPTH TDHS05-H3QPTH TDHS05-05QPTH TDHS05-L5QPTH TDHS05-0QPTH TDHS05-5QPTH TDHS05-0QGC 3.0 TDHS PTH TDHS05-L503PTH TDHS05-003PTH TDHS05-503PTH TDHS05-003GC 4.6 TDHS05-H346PTH TDHS PTH TDHS05-L546PTH TDHS05-046PTH TDHS05-546PTH TDHS05-546PTH TDHS05-004GC *Guard cartridge holder required, part no. XPGCH-Q Novel column hardware technology with increased pressure rating; specifications for the individual bonding chemistries remain identical irrespective of the column hardware format selected.

26 6 YMC-Triart YMC-Triart 63 Ordering Information YMC-Triart 3 µm analytical columns (max. pressure 00/50 bar) Phase C8 C8 Phenyl PFP Column ID (mm) Column length (mm) Guard cartridges* with 0 mm length (pack of 5).0 TAS03-00WT TAS03-030WT TAS03-050WT TAS03-L50WT TAS03-00WT TAS03-50WT TAS03-0QGC 3.0 TAS WT TAS03-L503WT TAS03-003WT TAS03-503WT TAS03-003GC 4.6 TAS WT TAS03-L546WT TAS03-046WT TAS03-546WT TAS03-546WT TAS03-004GC.0 TOS03-00WT TOS03-030WT TOS03-050WT TOS03-L50WT TOS03-00WT TOS03-50WT TOS03-0QGC 3.0 TOS WT TOS03-L503WT TOS03-003WT TOS03-503WT TOS03-003GC 4.6 TOS WT TOS03-L546WT TOS03-046WT TOS03-546WT TOS03-546WT TOS03-004GC.0 TPHS03-00WT TPHS03-030WT TPHS03-050WT TPHS03-L50WT TPHS03-00WT TPHS03-50WT TPHS03-0QGC 3.0 TPHS WT TPHS03-L503WT TPHS03-003WT TPHS03-503WT TPHS03-003GC 4.6 TPHS WT TPHS03-L546WT TPHS03-046WT TPHS03-546WT TPHS03-546WT TPHS03-004GC.0 TPFS03-00WT TPFS03-030WT TPFS03-050WT TPFS03-L50WT TPFS03-00WT TPFS03-50WT TPFS03-0QGC 3.0 TPFS WT TPFS03-L503WT TPFS03-003WT TPFS03-503WT TPFS03-003GC 4.6 TPFS WT TPFS03-L546WT TPFS03-046WT TPFS03-546WT TPFS03-546WT TPFS03-004GC YMC-Triart, 5 µm in ACTUS high-throughput semipreparative hardware (max. pressure 300 bar) Phase C8 C8 ExRS C8 Phenyl PFP Column ID (mm) Column length (mm) Ordering Information TAS05-050WX TAS05-00WX TAS05-50WX TAS05-50WX 30.0 TAS WX TAS05-L530WX TAS05-030WX TAS05-530WX TAS05-530WX 0.0 TAR08S05-050WX TAR08S05-00WX TAR08S05-50WX TAR08S05-50WX 30.0 TAR08S WX TAR08S05-L530WX TAR08S05-030WX TAR08S05-530WX TAR08S05-530WX 0.0 TOS05-050WX TOS05-00WX TOS05-50WX TOS05-50WX 30.0 TOS WX TOS05-L530WX TOS05-030WX TOS05-530WX TOS05-530WX 0.0 TPHS05-050WX TPHS05-00WX TPHS05-50WX TPHS05-50WX 30.0 TPHS WX TPHS05-L530WX TPHS05-030WX TPHS05-530WX TPHS05-530WX 0.0 TPFS05-050WX TPFS05-00WX TPFS05-50WX TPFS05-50WX 30.0 TPFS WX TPFS05-L530WX TPFS05-030WX TPFS05-530WX TPFS05-530WX HILIC.0 TDHS03-00WT TDHS03-030WT TDHS03-050WT TDHS03-L50WT TDHS03-00WT TDHS03-50WT TDHS03-0QGC 3.0 TDHS WT TDHS03-L503WT TDHS03-003WT TDHS03-503WT TDHS03-003GC 4.6 TDHS WT TDHS03-L546WT TDHS03-046WT TDHS03-546WT TDHS03-546WT TDHS03-004GC *Guard cartridge holder required, part no. XPGCH-Q YMC-Triart 5 µm analytical columns (max. pressure 00/50 bar) Phase Column ID (mm) Column length (mm) Guard cartridges* with 0 mm length (pack of 5) C8 C8.0 TAS05-00WT TAS05-030WT TAS05-050WT TAS05-L50WT TAS05-00WT TAS05-50WT TAS05-0QGC 3.0 TAS WT TAS05-L503WT TAS05-003WT TAS05-503WT TAS05-003GC 4.6 TAS WT TAS05-L546WT TAS05-046WT TAS05-546WT TAS05-546WT TAS05-004GC 0** TAS05-50WT TAS05-50WT TAS05-00CC.0 TOS05-00WT TOS05-030WT TOS05-050WT TOS05-L50WT TOS05-00WT TOS05-50WT TOS05-0QGC 3.0 TOS WT TOS05-L503WT TOS05-003WT TOS05-503WT TOS05-003GC 4.6 TOS WT TOS05-L546WT TOS05-046WT TOS05-546WT TOS05-546WT TOS05-004GC 0** TOS05-50WT TOS05-50WT TOS05-00CC.0 TPHS05-00WT TPHS05-030WT TPHS05-050WT TPHS05-L50WT TPHS05-00WT TPHS05-50WT TPHS05-0QGC Phenyl 3.0 TPHS WT TPHS05-L503WT TPHS05-003WT TPHS05-503WT TPHS05-003GC 4.6 TPHS WT TPHS05-L546WT TPHS05-046WT TPHS05-546WT TPHS05-546WT TPHS05-004GC 0** TPHS05-50WT TPHS05-50WT TPHS05-00CC.0 TPFS05-00WT TPFS05-030WT TPFS05-050WT TPFS05-L50WT TPFS05-00WT TPFS05-50WT TPFS05-0QGC PFP HILIC 3.0 TPFS WT TPFS05-L503WT TPFS05-003WT TPFS05-503WT TPFS05-003GC 4.6 TPFS WT TPFS05-L546WT TPFS05-046WT TPFS05-546WT TPFS05-546WT TPFS05-004GC 0** TPFS05-50WT TPFS05-50WT TPFS05-00CC.0 TDHS05-00WT TDHS05-030WT TDHS05-050WT TDHS05-L50WT TDHS05-00WT TDHS05-50WT TDHS05-0QGC 3.0 TDHS WT TDHS05-L503WT TDHS05-003WT TDHS05-503WT TDHS05-003GC 4.6 TDHS WT TDHS05-L546WT TDHS05-046WT TDHS05-546WT TDHS05-546WT TDHS05-004GC *Guard cartridge holder required, part no. XPGCH-Q (., 3, 4 mm ID) XPCHSPW (0 mm ID) **Max. pressure 00 bar