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A facile, preparative low-pressure chromatographic process was developed for purification of paclitaxel from crude extract of Taxus spp. The process consisted of normal-phase and a reversed-phase chromatography without utilization of any liquid-liquid partition step. The normal phase chromatography was designed lo separate paclitaxel from a majority of unwanted compounds to obtain an initial purification. Silica gel and Al2O3, were compared as packing materials for this step. The result demonstrated that basic Al2O3 was better than silica gel to give a higher throughput. A further advantage of the Al2O3 was its ability of removing l0-deacetyl-7-epipaclitaxel. a compound difficult to be separated from the paclitaxel in subsequent purification. The chromatographic fraction of A2O3 was purified by a subsequent reversed-phase column chromatography, employing a novel uniform porous microsphere, named PST, as the packing material. Combination of the two steps of column chromatography and one step of crystallization was able to purify paclitaxel, from 0.46% to more than 98% with the total recovery- of 71%. This integrated chromatographic procedure was reproducible, efficient, and simple to use. [ABSTRACT FROM AUTHOR]

A solid phase conjugation process was developed for attachment of polyethylene glycol to hemoglobin molecule. Bovine hemoglobin was loaded onto an ion exchange chromatography column and adsorbed by the solid medium. Succinimidyl carbonate mPEG was introduced in the mobile phase after the adsorption. Pegylation took place between the hemoglobin on the solid phase, and the pegylation reagent in the liquid phase. A further elution was carried out to separate the pegylated and the unpegylated protein. Analysis by HPSEC, SDS-PAGE, and MALLS demonstrated that the fractions eluted from the solid phase contained well-defined components. Pegylated hemoglobin with one PEG chain was obtained with the yield of 75%, in comparison to the yield of 30% in the liquid phase pegylation. The P50 values of the mono-pegylated hemoglobin, prepared with SC-mPEG 5kDa, 10kDa and 20kDa, were 19.97, 20.23 and 20.54 mmHg, which were much closer to the value of red blood cells than that of pegylated hemoglobin prepared with the conventional method. [ABSTRACT FROM AUTHOR]