Shimadzu pumps

General Experimental Procedures MPLC fractionation was performed using an Isco CombiFlash, and HPLC isolation was performed using Shimadzu pumps and detector and YMC-Pack ODS-AQ C18 column. 

Figure 13-10. Photograph of a Spark-HoUand system, which can be used in the off-line (LC) and on-Une SPE (XLC) modes. This nnit contains an autosampler, two Shimadzu pumps, a degasser, and SPE capability (for more details see www.sparkholland.com).

The recycle reactor is shown schematically in Figure 1. It consists of a catalytic or electrocatalytic reactor unit with a bypass loop, a recycle pump and a molecular sieve trap unit. The latter comprises one or two packed bed columns in parallel each containing 2-10 g of Linde 5A molecular sieve pellets. On line gas chromatography (Shimadzu 14A) was used for the analysis of CH4, O2, CO, CO2, C2H4 and C2H6 in the reactants and products. 

High Performance Liquid Chromatographic (HPLC) Analysis. A Waters HPLC system (two Waters 501 pumps, automated gradient controller, 712 WISP, and 745 Data module) with a Shimadzu RF-535 fluorescence detector or a Waters 484 UV detector, and a 0.5 pm filter and a Rainin 30 x 4.6 mm Spheri-5 RP-18 guard column followed by a Waters 30 x 3.9 cm (10 pm particle size) p-Bondapak C18 column was used. The mobile phase consisted of a 45% aqueous solution (composed of 0.25% triethylamine, 0.9% phosphoric acid, and 0.01% sodium octyl sulfate) and 55% methanol for prazosin analysis or 40% aqueous solution and 60% methanol for naltrexone. The flow rate was 1.0 mL/min. Prazosin was measured by a fluorescence detector at 384 nm after excitation at 340 nm (8) and in vitro release samples of naltrexone were analyzed by UV detection at 254 nm. 

Mobile phases employed for the separations are housed in a cartridge and delivered to the LC columns through a set of binary HPLC pumps (Shimadzu Corporation), as shown in Figure 6.2. The pumps provide a flow rate accuracy of 2% or 2 fiL (whichever is greater) in constant flow pumping mode, with a flow rate precision of 0.3%. A degasser (two channels internal volume of 195 /.d. /channel) is also housed in the pump module employed to minimize the occurrence of air bubbles. 

Recently, Chu et al. reported an ultra-fast LC/MS method for analysis of cytochrome P450 3A4 and 2D6 inhibition assaysd Testosterone and dextromethorphan were used as the specific substrates for CYP3A4 and CYP 2D6, respectively. LC/MS analyses were performed on a Sciex API 3000 mass spectrometer equipped with a Shimadzu LC-lOAdvp pump and a PE 200 autosampler. A Phenomenex Luna CIS (4.6x30 mm) column was used along with very steep gradients. Each sample analysis was completed in 0.5 min. 

Acrylate, propionate and acetate were separated by HPLC on a Benson carbohydrate column (30 cm by 0.6 cm i.d.) (Cnromtec, Fort Worth, FL). The HPLC system used a Waters UK6 injector and a Waters Model 6000A pump (Waters Associates, Milford, MA) with a Conductomonitor III Detector (Laboratory Data Control, Riviera Beach, FL) and a Shimadzu Data Processor. The solvent was 0.15 mM H2S04 at a flow rate of 0.5 ml/min. Typical retention times (min) were acrylate, 13.3 acetate, 12.3 propionate, 14.5. The detection limits for acrylate, acetate and propionate were 10, 30 and 40 pM respectively. 

The method recommended by Cooper et al [47] was applied for the analysis of the amino acids Asp, Glu, AAA, Asn, Ser, Tyr, Met, Val, Trp, Phe, lie, Leu and Lys. The HPLC-system consisted of an autosampler with injection valve (Pharmacia Biosystems), pump (Irica), degaser (Gastorr), thermostat (Julabo), precolumn and column (Waters), fluorescence detector (Shimadzu) and a PC (486 DX66)... 

Williams et al. used a high performance liquid chromatographic assay method for dipyridamole monitoring in plasma [71]. The HPLC system uses a Waters model 6000 A solvent delivery pump equipped with a U6K injector, a pBondapak C 9 column (30 cm x 39 mm 10 pm), and a Model 440 absorbance detector. The signal from the detector was quantified using a Shimadzu data processor and an Omni-Scribe recorder. A mobile phase flow rate of 1.5 mL/min was produced by a pressure of approximately 102 atm (1500 p.s.i.). The mobile phase was 50 50 mixture of acetonitrile and 0.01 M sodium phosphate in water (adjusted to pH 7). The absorbance reading of dipyridamole in methanol was made at 280 nm. 

LC-MS/MS System Used and Analytical Conditions A Shimadzu liquid chromatography system was used (Shimadzu Scientific Instruments, USA). The system consisted of a Shimadzu SCL-10A VP system controller with four LC-10AD VP pumps. A Perkin-Elmer series 200 autosampler, equipped with a cool-tray (Perkin Elmer, Norwalk, CT, USA) was used for sample injection. Moreover, a six-port switching-valve was implemented in order to switch between an extraction column and the analytical column. Samples were injected onto the extraction column (Oasis HLB column). After a short extraction/equilibration time of 0.3 min, the samples were eluted from the extraction column onto the analytical column (Symmetry C18) (Figure 1). 

Molecular weights were obtained by GPC in dichloromethane (1 mL/min) using a Waters pump model 6000, an injector (Rheodyne) and a refractive index detector (RID-6A Shimadzu), equipped with a PL gel 5)iim mixed-C linear column. The system was calibrated using polystyrene standards with low polydispersity. 

IR, UV, and H NMR spectra were obtained using Jasco FTIR-410, Shimadzu UV-2400PC, and Jeol JNM-GX270 (270 MHz) spectrometers, respectively. Size exclusion chromatography (SEC) was carried out on a Jasco GPC equipment consisting of a PU-980 pump, an RI-930, and a Shodex KF-806M column with polystyrene as a standard and tetrahydrofliran (THF) as an... 

Reliasil column (2.0 mm x 250 mm) on a Beckman 126 Pump Module with a Shimadzu (SPD-6A) detector (Figure 1). The column was eluted for 2 min with solvent A (0.1% trifluoroacetic acid in water) and then followed by a discontinuous gradient to solvent B (10% methanol. 10% water, 80% acetonitrile) at a flow rate of 0.15 ml/min at 35°C. The gradient used was as follows ... 

The water-insoluble portion was then washed with 10 mL of MeOH to separate MeOH-soluble and MeOH-insoluble portions. The obtained water-soluble and MeOH-soluble portions, the sum of which nearly corresponds to the supercritical water-soluble portion, were then analyzed by the high perfonnance liquid chromatograph (HPLC) (Shimadzu LC-lOA) which consists of a high pressure pump (Shimadzu Co., Model LC-IOAT). HPLC analysis conditions were as follows [Column SHI ODS-11, Column temperature 40 6. Carrier solvent CHjOH/HjO = 20/80 (O- -IO min), 20/80 - 100/0 (10 20 min), 100/0 (20- 30 mm), Flow rate 0.7 ml/min. Detector a spectrophotometric detector (SPD) (A =254 nm) or a refractive index detector (RID)], or [Column ULTRONPS-80P, Column temperature SOTl. Carrier solvent H2O, Flow rate 1,0 ml/min, Detector SPD (A =254 nm) or RID], To obtain the yields of monosaccharides and their decomposed products, their standard samples (Nacalai tesque, extra pure reagent) with known concentrations in water were analyzed by HPLC as a standard in a similar manner. 

To study the molecular distribution of water-soluble and MeOH-soluble portions, gel permeation chromatography (GPC) (Shimadzu LC-IOA) which consisted of a high pressure pump (Shimadzu Co, Model LC-IOAT) was performed with a column of KF-803L, operated at 401 with flow rate of 1.0 mL/min of tetrahydrofuran, The molecular weight of the fractions was determined from calibration curve obtained with polystyrene as a standard. 

SC-CO9 Purification of Reaction Mixture For purification of smaller batches (20 g) of the crude reaction mixture, a scaled down version of the large vessel, a 6-foot length of 1.0 OD x 11/16 ID SS pipe, was used. In addition, several purifications were performed using 4 + 0.5 wt% ethanol (EtOH) as a cosolvent. The EtOH was injected at a constant volumetric flow rate into the preheater where indicated in Figure 1 using a Shimadzu LC-6A HPLC pump. 

The catalytic reactions were carried out at atmospheric pressure in a conventional flow reactor using a U-shaped quartz tube (0 10 mm) with a fixed bed catalyst. The catalyst was diluted with quartz beads. CH4 and H2O were mixed with N2 in the ratio of 1/2/2. The flow rate of CH4 and N2 was controlled with a mass flow controller (STEC SEC-400 Mark3). Distilled water was fed into the reactor with a liquid pump (Shimadzu LC-lOADvp) through a vaporizer. The space velocity changed from 6.0 x 10 to 3.0 x 10 ml h g-cat . The products were analyzed by three on-line TCD gas chromatographs with Porapak-Q and Molecular Sieve 5A columns. 

Methanol oxidation was carried out in a conventional flow apparatus at atmospheric pressure. The feed mixtures were prepared by injecting the liquid methanol into air flow with a Gilson 302 pump. The catalyst was diluted with inert carborundum (1 3 volume ratio) to avoid adverse thermal effects, and placed in a tubular pyrex reactor with a coaxially centred thermowell with thermocouple. The reactor outlet was kept at 403 K, to prevent condensation of liquid products and formaldehyde polymerization, and it was connected with multicolumn Shimadzu GC-8A gas chromatograph with thermal conductivity detector. The column system used (1.5m of Poropak N+1.5m of Poropak T+0.9m of Poropak R) could separate CO2, formaldehyde, dimethylether, water, methylformate, dimethoxymethane and formic acid. The last product was never detected. 

The HPLC-PO-CL system (Fig. 2) consisted of two LC 9A liquid chromatographic pumps (Shimadzu, Kyoto, Japan), a Rheodyne 7125 injector (Cotati, CA, USA) with a 20- tL sample loop, an ultraviolet lamp, Toshiba GL-10 (10 W, 254 nm), a Chemcosorb 5-ODS-UH column (150 x 4.6 mm I.D.), CLD-lOA detector (Shimadzu), a Rikadenki R-61 recorder. PTFE tubing (6.0 m x 0.5 mm I.D., GL Sciences, Tokyo) coiled around the ultraviolet lamp as the on-line for the UV radiation reactor. 

The separation of DIB derivatives of morphine and IS were performed using an HPLC system (Shimadzu, Kyoto, Japan) consisting of two pumps (LC-lOATvp) with a system controller (PX-8010), a recorder (FBR-2), a FL detector (RF-550) set at Xex=355 nm and Xem=486 nm, and a Rheodyne 7125 injector (Cotati, CA, USA) with a 20-pL sample loop. In plasma analysis, the mobile phases used were a mixture of acetonitrile-0.1 M acetate buffer (pH5.4) (50 50, v/v, MPl) with a flow rate of 1.0 mL/min and acetonitrile (MP2). The separation program was set as follows the flow rate of MP2 was set at 0 mL/min from 0 to 29 min, rapidly... 

The FIA system consisted of two CCPD chromatographic pumps (Tosoh, Tokyo, Japan), an 825-CL detector (Jasco, Tokyo), a 7125 injector with a lOO-pL of sample loop (Rheodyne, Cotati, CA,USA) and an R-111 recorder (Shimadzu, Kyoto, Japan). 

The HPLC system consisted of five LC-lOA pumps, an SIL-lOA auto sample injector, a DGU-14 degasser, a CLD-lOA chemiluminescence detector, an SCL-lOA system controller, a C-R4A integrator, an HIC-6A and a CTO-IOAC column oven (all Shimadzu, Kyoto, Japan). The clean-up colunm (4.6 i.d. X 150 mm), concentrator column (4.6 i.d. X 30 mm), separator colunm (4.6 i.d. X 250 + 150 mm), guard columns 1 (4.6 i.d. X 30 mm) and 2 (4.6 i.d. X 50 mm) were packed with Cosmosil 5C18-MS (Nacalai Tesque, Kyoto, Japan), and the reducer column (4.0 i.d. X 10 mm) was a Nitroarene Reactor Column (Shimadzu). All other conditions were the same as those given in our previous report. ... 

Major manufacturers of HPLC instruments include Waters, Agilent (formerly Hewlett Packard), and Shimadzu, PerkinElmer, Thermo, Beckman, Varian, Hitachi, Jasco, Dionex, Gilson, Scientific Systems (SSI), and Isco. The Internet addresses of these companies can be found in the reference section. HPLC is a mature technology and most manufacturers have highly reliable products with sufficient performance and feature sets to be competitive in the market place. However, there can still be significant differences between the vendors on these performance characteristics on systems (dwell volume, dispersion), pumps (low flow, seal life), autosamplers (carryover, speed, sample capacity, minimum sample volume), and detectors (sensitivity, gradient baseline shift). 

Analyses were performed by HPLC using a Shimadzu LC-6A pump and UV Spectrophotometer SPD-6A detector. The columns used wei e Phenomenex Rezex monosaccharide-H+ with trifluoroacetic acid (lO M) as the mobile phase for toluene as the solvent, and Spherisorb-CN with cyclohexane/methylene chloride/isopropanol (80/16/4 by volume) as the mobile phase for methyl isobutyl ketone as the solvent. 

---