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HPLC Peptide Standards for RP on the HAISIL HL C18 Column

The aromatic hydrocarbons that most HPLC column manufacturers use to quality control columns render little information about a column's selectivity

and performance characteristics for peptides.The Canadian corporation, SPI, produces a well characterized yet challenging peptide mixture that is very useful for quantifying HPLC column performance for peptides.The difficult to separate first pair of peptides are resolved to near base line on HAISIL HL C18 columns. Many so-called peptide analysis columns are unable to separate this pair.

HA I S I L HL A P P L I C AT I ONS

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Speed and Selectivity for Ballistic Gradients on Complex Libraries

The relative impor tance of par ticle size and selectivity are graphically illustrated below by the high speed analysis of a six component combinatorial library on a HAISIL HL C18 5µm column and two competitor’s C18 2µm columns. HAISIL HL with its high carbon loading and surface area of often works much better than expensive small par ticle size columns that are prone to plugging up more quickly.

Competitor “A” 2µm C18

HAISIL HL 5µm C18

Competitor “B” 2µm C18

Water Soluble Vitamins

HAISIL HL C18 5µm 250x4.6mm A: 0.05mM KOAc, pH5; B: MeCN Gradient: 2.5% - 30%B in 25min

Food Additives

Beverage additive analysis is not only an impor tant industrial application, but it also serves as a practical and relatively simple procedure for introductory HPLC courses. Industry is increasingly driven by rapid analysis requirements and an HPLC column can represent an overwhelming expense in a teaching environment.The diet cola analysis in the chromatogram to the left illustrates an extremely fast analysis on an extremely inexpensive HPLC column. A package containing five

of the 20x2.1mm car tridge column used for this analysis costs about half as much as one moderately priced HPLC column. And, there's no need to pre-filter the cola sample through a 0.5µm filter as some column manufacturers recommend when using their expensive columns packed with 2µm particles for this analysis! The readily available and non-toxic nature of the sample and the low cost of Higgins Analytical cartridges has made the diet cola analysis an ideal choice for classroom purposes.

Basic Compounds in Combinatorial Mixtures

HAISIL HL C18 P/N LS-1546-C185

Reverse Phase Chromatography of Basic Compounds in Combinatorial Mixtures

In the example at the top of the next column, A and B are single compounds containing two identical building blocks. A-B and B-A represent positional isomers, each containing two building blocks.

This analysis was conducted on the crude liquid-phase reaction mixture. Each component contains multiple secondary

and tertiary amine functionality yet the combinatorial products can be rapidly chromatographed and exhibit no tailing on a 150x4.6mm HAISIL HL C18 column using a simple linear 5% to 100% MeCN/water 0.1% TFA gradient.

Water Soluble Vitamin Analysis on HAISIL HL

1. H2N-RGAGGLGLGK-amide 2. AC-RGGGGLGLGK-amide 3. AC-RGAGGLGLGK-amide 4. AC-RGVGGLGLGK-amide 5. AC-RGVVGLGLGK-amide

“Hodges Test Peptide Mix” (from Synthetic Peptides, Inc.)

HAISIL HL C18 5µm 150x4.6mm P/N LS-1546-C185

Fast Diet Cola Analysis

HAISIL HL C18 5µm 2cm x 2.1mm Cartridge PN LK-0221-C185 15% MeCN/Water 200µL/min

Caffeine

Aspartame

Sodium Benzoate