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than that of desired product. Collection plates have a 2 mL deep-well microtiter format and are custom designed with expansion chambers to accommodate evaporation of carbon dioxide as it departs. A 31-second timeout at the ow rate of 12 mL/min is used to avoid collection above a desired volume per sample and to achieve collection of samples into single wells. In-house designed software tracks all compounds collected with respect to plate and well location. 11.1.5. Abbott Laboratories HTP Scope
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The high-throughput puri cation (HTP) group at Abbott Laboratories is based strictly on high-performance chromatography techniques so as to yield purities at 95% and above. Further HTP is set up as a service function rather than as open-access instrumentation. The synthetic chemists drop off samples at a centralized location, the puri cation work is carried out by a staff of puri cation chemists dedicated to the task, and puri ed compounds are returned to the synthetic chemist along with structural validation based upon MW. A wide range of synthetic formats are submitted to this HTP service, including those from a high-throughput organic synthesis (HTOS) group (who synthesize, on a service basis, 48 member libraries), a combinatorial chemistry projects support (CCPS) group dedicated to speci c therapeutic targets and synthesize highly variable-sized libraries (from ten to hundreds of members), as well as numerous medicinal chemists, who typically synthesize single or small numbers of compounds of a particular class at a time. Similarly the weights of materials synthesized by these three distinct groups of synthetic chemists vary as do their library sizes. Whereas the HTOS group generate strictly 10 to 50 mg member libraries, the CCPS libraries may vary from 10 to 100 mg and the medicinal chemists single samples from 50 to 300 mg. This high variance in library size and entity weight requires a highly exible puri cation service. Additionally each of the three synthetic schemes presents speci c puri cation challenges. HTOS libraries, based on standardized chemistries, tend to give a high variability in product yield. The CCPS libraries are often carried out on valuable core in late-stage development, and hence reliability in return of the puri ed material is of paramount importance. Compounds synthesized by the medicinal chemist are submitted to HTP in sets of one, two, or few reaction mixtures and hence present a challenge in terms of processing ef ciency. The scope of the present HTP service is capable of processing samples sized from 10 to 300 mg in weight, maintains for the client chemists a 72 hour turnaround, has the capacity to purify 25,000 samples a year, and operates with a 99% success rate. To maintain this broad charter of service, it is
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high-throughput puri cation: triage and optimization
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required for submission of samples to HTP that analytical proof be provided for the presence of desired component via an analytical HLPC/MS trace, and that the desired component of the crude reaction mixture be present in at least an estimated 5% yield. Additionally the puri cations requested must be of the routine type, excluding natural products mixtures, no proteins, large peptides, or highly insoluble (in DMSO or MeOH) materials.
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PURIFICATION SYSTEMS AND ANALYTICAL SUPPORT
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To accommodate the variant needs of the different synthetic chemistry groups, a series of high-performance chromatographic systems are employed, including UV-triggered HPLC, ELSD-triggered HPLC, MWtriggered HPLC, and UV-triggered SFC. Each sample set submitted to the HTP service is directed to the speci c instrument and technique that best meets the puri cation requirements of the library, based on considerations of ef ciency and compatibility of the chromatographic technique to the structures, weights, and puri cation complexity. HPLC (UV/ELSD) The puri cation system rst set up for our HTP service is a series of UV/or/ELSD-triggered fraction collection HPLC instruments.26 Each UV/ELSD-HPLC is comprised of a Waters (Milford, MA) PrepLC 4000 solvent delivery and control system, a Waters 996 Photo PDA detector, Waters 717 plus auto-sampler, an Alltech (Deer eld, IL) Varex III ELSD, and two Gilson (Middletown, WI) FC204 fraction collectors. The chromatography columns used are Waters SymmetryR columns in 7-m particle size, radial compression format, employing different sizes: 10 100 mm, 25 100 mm, and 40 100 mm for the puri cation of samples sized from 10 20, 20 70, and 70 300 mg, respectively. The standard gradient elution methods used are 0 100% CH3CN : 0.1% aqueous TFA or ammonium acetate, 10 100% CH3CN : 0.1% aqueous TFA or ammonium acetate, 20 100% CH3CN : 0.1% aqueous TFA or ammonium acetate, and 50 100% CH3CN : 0.1% aqueous TFA or ammonium acetate. The fraction collection is triggered by PDA detection at 220, 240, or 254 nm or by ELSD, based on an evaluation of analytical LC/MS data. To ensure return to the synthetic chemist of the desired product or products from the UV- or ELSD-triggered fraction collection, loop injection
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