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High-Performance Liquid Chromatography. Advances and by Csaba Horvath

By Csaba Horvath

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Phenylboronic acid as a group-specific ligand reversibly forms cyclic boronate esters with the boronate anion at high pH and thus is well suited for the resolution of diol-containing molecules such as nucleosides, nucleotides, carbohydrates, and RNA (Glad et al, 1980). 8 g of 3-aminobenzeneboronic acid hemisulphate dissolved in a minimum volume of water (an ultrasonic bath was used to facilitate the dissolution). The total volume of the mixture was 32 ml. 0 with 5 M sodium hydroxide. The coupling reaction was conducted at 21°C for 20 h.

First, the pore size of the packing becomes a dominant feature and this will necessarily lead to large pore-size packings with a pore diameter pd > 30 nm, compared to the 6- to 15-nm pore-size silicas in HPLC of low-molecular-weight compounds. It is very likely that future packings in thisfieldwill have a pore size of pd » 30 nm or may even become free of pores. With regard to the particle size, two trends become apparent; microparticulate silicas of a mean particle size dp < 5 μηι, packed in short columns for fast analysis, will be one route.

Columns have lengths up to 250-300 mm and inner diameters between 4 and 10 mm. Most of the SEC packings have particle sizes in the 10-μηι size range except the two DuPont Zorbax Bio Series Gel Filtration columns GF 250 (dp = 4 - 5 μπι) and GF 450 (dp % 6 μιη). In order to improve the pH stability of these two TABLE VII Commercial Silica-Bonded Affinity Packings Name Supplier Bakerbond MA b Bakerbond ABX Baker Baker Bakerbond Ultraprep Affinity Matrices (activated silica supports) Affinity Matrices (activated silica supports) Affinity Matrices (activated silica supports) Ultraffinity-EP CPG/CDI-Activated Glycophase CPG/Aminoacryl CPG/Aminopropyl CPG/Carboxyl CPG/Long-Chain Alkylamine CPG/NHS Glycophase CPG/Stable Diazonium Salt CPG/Thiol Baker Baker Baker Baker Baker Beckman Pierce Pierce Pierce Pierce Pierce Pierce Pierce Pierce a Form 0 Mean particle diameter dp (μιτι) Nominal pore size PA (nm) o, x o, x 5,15,40 5,15,40 28,30 28,30 X 40 40 40 40 40 10 74-125 125-177 125-177 125-177 125-177 125-177 125-177 125-177 28 28 28 28 28 — X X X — x,b x,b x,b x,b x,b x,b x,b x,b Abbreviations: o, spherical particles; b, bulk material; x, irregular particles; —, not given.

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