ACE C8-Amide For increased polar retention and alternative selectivity Alternative selectivity for method development Improved separations with polar, acidic, basic and phenolic compounds High efficiency µm, µm, µm and 0µm particles for UHPLC and HPLC Ultra-inert for maximum performance and reproducibility ACE ACE C8-Amide for increased polar retention and UHPLC alternative and HPLC Columns selectivity
ACE C8-Amide For increased polar retention and alternative selectivity Ideal column choice for method development Alternative selectivity to C8 and C8 columns with polar molecules - especially for acids Compatible with 00% aqueous mobile phases High efficiency µm, µm, µm and 0µm particles for UHPLC, HPLC and preparative separations Low bleed for UV and LC/MS compatibility Recommended Applications Small water soluble analytes and polar compounds H-bond donors, acids, bases and phenolic compounds Small peptides Why does ACE C8-Amide Provide Alternative Selectivity? ACE C8-Amide combines a C8 with a polar amide group on a single ligand Extended spacer technology additionally provides extended column lifetime C8 Phase ACE C8-Amide C8 group for hydrophobic interaction C8 group for hydrophobic interaction H N 0 amide group increases retention for polar compounds and provides alternative selectivity extended spacer technology for increased lifetime and stability R Si R R Si R ultra-inert, ultra pure, ultra reproducible ACE silica Hydrophobic interaction only ACE C8-Amide provides multiple modes of interaction ACE C8-Amide for increased polar retention and alternative selectivity
Leverage the Power of Selectivity with ACE C8-Amide to pull peaks apart The resolution equation determines the parameters that contribute to resolution; N, α and k. In recent years there has been a significant focus on the use of ultra efficient UHPLC columns (such as ACE Excel µm columns) as a means of achieving separation goals. Resolution Efficiency Selectivity Retention Factor However, selectivity is often overlooked. This is unfortunate as of the three parameters that affect resolution, selectivity is the most powerful (see Figure ). By leveraging both efficiency and selectivity, better and faster separations can often be achieved. ACE C8-Amide is the latest addition to the ACE range of bonded phases which offer both complementary selectivity and the option of µm, µm, µm and 0µm particle sizes - to enable development of rugged, robust UHPLC and HPLC methods. Leverage the power of bonded phase selectivity to pull peaks apart Figure illustrates the difference between two ACE bonded phases, C8-Amide and C8. Although both phases offer the possibility of strong hydrophobic interaction from their respective C8 chains, the amide group embedded within the C8-Amide phase introduces additional modes of interaction which ultimately increases retention for polar compounds and provides alternative selectivity. Fig. - The Effect of N, α and k on Resolution (R s ) For a typical separation where N = 0,000, k = and α =. Fig. - Use ACE C8-Amide to Enhance Selectivity Application # 0 ACE Excel C8-Amide Enhanced selectivity due to multiple modes of interaction from the ACE C8-Amide ligand Resolution (R s ) 0 0 ACE Excel C8, Increasing N, α or k increases resolution (R s ), but as seen from the above plots, increasing either N or k suffers from quickly diminishing returns. Increasing selectivity (α) does not have this problem and is therefore the most powerful of these three variables to optimise when developing a separation. C8 phase interacts only with a hydrophobic mechanism 0 0 Sample: ) resorcinol ) catechol ) -methyl resorcinol ) -methyl catechol ) -methyl catechol ) -nitro catechol Mobile Phase: : MeCN/mM H PO in H O Column Dimensions: 0 x.mm Flow Rate:.0ml/min Temperature: 0 C Wavelength: 0nm ACE C8-Amide for increased polar retention and alternative selectivity
ACE C8-Amide Provides Enhanced Polar Selectivity Fig. - Comparison of ACE C8-Amide to Leading C8 UHPLC Columns Application # 0 Reproduced with kind permission of The Open University, UK ACE Excel C8-Amide (ultra-inert fully porous silica) ACE C8-Amide provides increased retention and improved separation due to enhanced polar selectivity 0 0 ACE Excel C8 (ultra-inert fully porous silica) 0, Waters Acquity. BEH C8 (hybrid particle) 0, Phenomenex Kinetex. C8 (superficially porous particle) Leading C8 brands provide similar selectivity 0,, Thermo Accucore. C8 (superficially porous particle) 0, Zorbax Eclipse.8 XDB C8 (fully porous silica) Column Dimensions: 0 x.mm Sample: ) resorcinol ) catechol ) -methyl resorcinol ) -methyl catechol ) -methyl catechol ) -nitro catechol Mobile Phase: : MeOH/mM H PO in H O Flow Rate: 0.0ml/min Temperature: 0 C Wavelength: nm 0 Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks. ACE C8-Amide for increased polar retention and alternative selectivity
Exploit Alternative Selectivity for Method Development Fig. - Separation of Acidic, Basic and Neutral Compounds Application # 0 8 ACE Excel C8-Amide ACE C8-Amide phase offers complementary selectivity to C8 columns due to multiple modes of interaction from the unique C8-Amide ligand 0,, 8 ACE Excel C8, 0 Sample: ) methylphenylsulphoxide ) pindolol ) -hydroxybenzoic acid ),-dimethoxybenzene ) berberine ) myricetin ) piperine 8) chrysin Mobile Phase: A = 0mM ammonium formate in H O (ph.0) B = 0mM ammonium formate (ph.0) in 90:0 (v/v) MeOH/H O Gradient: 00% B in minutes Column Dimensions: 0 x.mm Flow Rate: 0.0ml/min Temperature: 0 C Wavelength: nm Compatible with 00% Aqueous Mobile Phases Fig. - Separation of Organic Wine Acids Application # 0 ACE C8-Amide is compatible with 00% aqueous mobile phase for maximum retention and resolution Conventional alkyl C8 columns are not recommended with 00% aqueous conditions due to irreproducible retention loss caused by pore dewetting 0 8 Sample: ) oxalic acid ) tartaric acid ) malic acid ) lactic acid ) ascorbic acid ) citric acid Mobile Phase: 0mM NH H PO in H O (ph.) Column: ACE Excel C8-Amide, 0 x.mm Flow Rate: 0.ml/min Temperature: C Wavelength: nm ACE C8-Amide for increased polar retention and alternative selectivity
ACE C8-Amide Provides Alternative Selectivity Fig. - Food and Beverage Additives - Vanillins Application # 0 ACE Excel C8-Amide Improved resolution due to multiple modes of interaction from the ACE C8-Amide ligand 0 8 0 ACE Excel C8, C8 phase interacts only with a hydrophobic mechanism 0 8 0 Sample: ) vanillic acid ) -hydroxybenzoic acid ) vanillin ) -hydroxybenzaldehyde ) guaiacol ) ethyl vanillin ) eugenol Mobile Phase: A = 0.% formic acid in H O B = 0.% formic acid in MeCN Gradient: 0 % B in 0 minutes Column Dimensions: 0 x.mm Flow Rate:.00ml/min Temperature: 0 C Wavelength: 0nm Fig. - Steroids Application # 0 ACE Excel C8-Amide ACE C8-Amide provides increased retention and improved separation due to enhanced polar selectivity 8 9 0 0 8 0, ACE Excel C8 8 9 0 0 8 0 Sample: ) prednisone ) prednisolone ) estriol ) corticosterone ) a-hydroxyprogesterone ) -ketoprogesterone ) -hydroxyprogesterone 8) β-estradiol 9) a-estradiol 0) a-ethynylestradiol Mobile Phase: A = 0.% formic acid in H O B = 0.% formic acid in MeCN Gradient: 80% B in 0 minutes Column Dimensions: 0 x.mm Flow Rate: 0.0ml/min Temperature: C Wavelength: 0nm ACE C8-Amide for increased polar retention and alternative selectivity
Guaranteed Reproducibility and Fully Scalable Fig. 8 - Reproducible Scale Up with ACE C8-Amide Application # 0 ACE Excel C8-Amide LC/MS 0 x.mm 0.ml/min (Silane Batch #) 0 8 9 0 ACE Excel C8-Amide Analytical 0 x.0mm 0.ml/min (Silane Batch #) 0 8 9 0 ACE Excel C8-Amide Analytical 0 x.mm.00ml/min (Silane Batch #) 0 8 9 0 ACE 0 C8-Amide Preparative 0 x.mm.ml/min (Silane Batch #) 0 8 9 0 Sample: ) uracil ) -hydroxybenzoic acid ) acetylsalicylic acid ) benzoic acid ) -hydroxybenzoic acid ) ethyl paraben Mobile Phase: : MeCN/0.% TFA in H O Temperature: C Wavelength: nm The availability of µm, µm, µm and 0µm particle sizes combined with a range of dimensions from UHPLC through to preparative HPLC scale ensures that methods can be reproducibly scaled up or down. The chromatograms in figure 8 demonstrate the excellent reproducibility achieved when changing both silica batch and silane batch, and the reproducible scalability obtained when changing particle size and column diameter. ACE C8-Amide for increased polar retention and alternative selectivity
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