Waters Alliance HPLC

FIGURE 16 The chromatogram of an injection of a caffeine solution without the column showing the instrumental bandwidth of a Waters Alliance HPLC system with a 966 PDA detector with a standard flow cell. 

Figure 1 Separation of a chromatographic test mixture by a traditional gradient method. Separation was performed on Waters Alliance HPLC System (Waters Corporation, Milford, MA) and a 3.9 by 150mm 5 micron particle size Symmetry Cl8 column at 30°C. The mobile phase consisted of 0.1% phosphoric acid as the A solvent, and acetonitrile as the B solvent, run as a linear gradient from 0-80% B over 40 minutes at 1.0 mL/min. Total analysis time does not include twenty minutes of post run reequilibration of the column and system. UV detection at 254nm, and a 20 pL injection was used. Peaks 1-12 (O.lmg/mL each in 50/50 methanol/water) are uracil, theophy-line, acetylfuran, acetanilide, acetyl-, propio-, butyro-, benzo-, valero-, hexano-, hep-tano-, and octano-phenone, respectively.

Products/technologies The Waters LC-MS solutions for drug discovery consist of a Waters Alliance HPLC system (with optional 996-photodiode array or 2487 UV/Vis detectors), Waters 2700 Sample Manager (for 96-well microtiter plates), Micromass Platform LC or Platform LCZ benchtop API mass spectrometer, and MassLynx software with special options for combinatorial... 

Automated Method Waters Alliance HPLC with PDA 65,000-... 

Software ChromSword 3.1, Waters Empower. Waters Alliance HPLC system. 

Figure 3.20 shows the separation of a mixture on four HSS C18 columns with different particle sizes and dimensions (53). The column packed with sub-2 pim particles was run on Waters Acuity UHPLC, and the other three conventional columns were run on Waters Alliance HPLC systems. The difference in Vdweii/Vo between each set up was compensated by changing the length of initial gradient hold, and... 

A Waters 2690 Alliance HPLC equipped with a 996 photodiode array and a 896 IJV/Vis detector was used for carotenoid analysis. The column (Phenomenex, Torrance, CA) was a 250x4.6mm Ultracarb 3 pm C-18 stationary phase and elution was carried out isocratically at a flowrate of l.OmL/min with 85 15 (v v) acetonitrileimethanol (HPLC grade) containing 0.1% triethyl amine to prevent on-column carotenoid decomposition. 

FIGURE 2 Examples of modular and integrated HPLC systems. Upper row (I to r) modular systems Agilent 1100 Series, Jasco LC-2000 Series Lower row (I to r) integrated systems Waters Alliance, Shimadzu LC-2010. 

Separation of five compounds (DL, 6-OH-DL, 3-OH-DL, 7V-OH-DL, and 1-pyridine-/V-oxide-DL) was achieved using an Alliance HPLC system (Waters Corp., Milford, CA) equipped with a 2690 model pump, an autoinjector, a Polaris Cl8-A guard column (Varian Inc., Lake Forest, CA), and a Luna Phenyl-Hexyl analytical column (Phenomenex, Inc., Torrance, CA) maintained at 40°C. For robust characterization of each isomeric compound, an online HDX LC-MS method was developed. The composition of regular and deuterated mobile phases is summarized below ... 

Figure 4.2. Examples of modular and integrated HPLC systems. Modular systems (a) Agilent 1100 Series, (b) Jasco low-pressure mixing system. Integrated systems (c) Waters Alliance, (d) Shimadzu 2010.

HPLC-MS analyses were carried out with a Waters Alliance 2690 HPLC (Milford, MA), with column heater, 996 Photodiode Array Detector, and a Micromass ZMD (Waters, Milford, MA) or Waters Integrity TMD mass spectrometer (Milford, MA). A Luna silica-2, 5 pm, 2.0 x 150 mm (Phenomenex, Torrance, CA) column was used at a temperature of 35°C. The ZMD MS was equipped widi an atmospheric pressme chemical ionization (APCI) source and parameters set as corona voltage, 3.5 kV cone voltage, 30 V source block temp, 125 C APCI heater, 400 C desolvation gas, 150 L/min cone gas flow, 100 L/min scan range, m/z 151-1000. Reversed phase LC-MS was performed using a Beckman HPLC with 126 pump, 168 PDA (Fullerton, CA), and a Thermoquest LCQ mass spectrometer (San Jose, CA). A Luna Cl8, 3 pm, 2.0 X 50 mm (Phenomenex, Torrance, CA) column was used at ambient temperature. The LCQ MS was equipped with an APCI source and parameters set as APCI vaporizer temp, 450 C source current, 5 pA sheath gas flow, 80 au aux gas flow, 20 au capillary temp, 150 C capillary voltage, 23 V scan range, m/z 250-1500. 

Waters Alliance Systems HPLC w/RI -I- UV detectors HPLC oriented... 

America, San Jose, CA, USA, and Scientific Software Inc., Pleasanton, CA, USA, versions for Hitachi LaChrom and LaChromElite hardware and EZChromElite software were launched. Partnership with Agilent Technologies resulted in the creation of a powerful version that supports Agilent 1100 LC and LC/MS systems with six columns and twelve solvent selectors. Further systems supported by ChromSword Auto are Waters Alliance LC systems with Millenium and Empower softwares. ChromSword Auto also supports 2-8 column and 2-16 solvent switching valves for all HPLC systems described. 

Their method included a Waters 2795 Alliance HT (high throughput) HPLC system with an integrated autosampler. The stationary phase was a Supelco C18 column (250 x 4.6 mm, 5 fim). The mobile phase consisted of solvent A (water containing 2mM ammonium acetate and 0.1% formic acid) and solvent B (methanol containing 2mM ammonium acetate and 0.1% formic acid). The mobile phase was delivered at a flow rate of 0.6 mL/min in a step gradient mode 50% solvent B from 0 to 0.4 min and 100% solvent B from 0.4 to 0.8 min. 

The concentrations of the individual silymarin compounds and taxifolin were determined by HPLC using a Waters system (Milford, MA) composed of an Alliance 2690 separations module and a 996 Photodiode Array detector controlled with Millennium32 chromatography software. The HPLC procedure was previously described by Wallace et al. (13). Calibration curves were prepared from standard solutions of taxifolin, silychristin and silymarin. The silybinin standard from Sigma contained two distinct peaks, which are further referred to as silybinin A (the first peak) and silybinin B (the second peak). 

Identify the peptides with high performance liquid ehromatography (HPLC) on an Alliance 2690 from Waters (Milford, MA) and eleetrospray mass speetrom-etry (ESI-MS) on a Platform II from Micromass (Manchester, UK). 

The HPLC system, an Alliance 2790, with thermostatted autosampler and divert valve LabPro, UV detector 2487 and satin interface, and ZQ mass spectrometer with ESI interface, were products of Waters Inc., Milford, Massachusetts. The chemiluminescent nitrogen detector (CLND) model 8060 was a product of ANTEK Instruments Inc., Houston, Texas. The liquid handler Miniprep 75 was a product of Tecan Group Ltd., Maennedorf, Switzerland. 

Concentration of antioxidants in chemical reagents was analyzed by RP-HPLC method. The RP-HPLC system consisted of a Waters 2695 Separations Module (Alliance Waters, Milford, USA), Waters 2996 PDA Detector, Waters 2487 Dual X Absoibance Detector, and a column of length 150 mm x 4.6 mm ID, 5 pm particle size. RP-HPLC analysis conditions (See Table 1). 

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