A series of parameters, including composition and pH of mobile phase, column packing, flow rate and detection wavelength, were tested with respect to the location and shape of the peak of acyclovir in the corresponding chromatograms. The final choice of the stationary phase giving satisfying resolution and run time was a reverse phase Hibar LiChrospher 100 RP8, 250 × 4.6 mm I.D. (5 μm, particle size), protected by a guard column LiChrospher RP8 4-4 mm (5 μm). A mobile phase consisting of 100 % water solution of 0.1 % (V/V) triethylamine with pH = 2.5 adjusted with concentrated ortho-phosphoric acid delivered by a pump Perkin Elmer LC series 200 was found to give best results. The mobile phase was filtered and degassed with helium. A flow rate was 1.2 ml/min. Chromatographic separations were performed at 25°C. An ultraviolet diode array detector (Perkin Elmer LC 235 C) was used for detection and 255 nm was chosen as optimal for determination of acyclovir. The samples were introduced in the column using an autosampler Perkin Elmer LC ISS Series 200 and the injection volume was 120 µl. The chromatographic system was controlled by the software package Turbochrom Version 4.1. plus and UV-spectrometric data were produced by TurboScan Version 2.0.
Series 200 lc
Manufactured by PerkinElmer
Sourced in United States, United Kingdom
The Series 200 LC is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a modular design, allowing for the integration of various components such as pumps, autosamplers, and detectors to create a customized HPLC solution. The Series 200 LC provides precise and reliable separation and quantification of a wide range of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Lc iss series 200, by PerkinElmer (2 mentions)
Lc 235 c, by PerkinElmer (2 mentions)
Analyst software v1, by AB Sciex (2 mentions)
Securityguard c18 column, by Phenomenex (2 mentions)
Api4000 triple quadrupole lc ms ms, by Thermo Fisher Scientific (2 mentions)
Luna c8 2 column, by Phenomenex (2 mentions)
Avanti jxn 26 centrifuge, by Beckman Coulter (1 mentions)
Sonic 5, by Polsonic (1 mentions)
Microwave 3000, by Anton Paar (1 mentions)
Hplc system, by PerkinElmer (1 mentions)
Iss200, by PerkinElmer (1 mentions)
Fc203b, by Gilson (1 mentions)
9 protocols using series 200 lc
1
Optimized HPLC Assay for Acyclovir
2
Diazepam Quantification by HPLC-UV
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The development and validation work was carried out on a chromatographic system consisting of Perkin Elmer LC series 200 pump, ultraviolet diode array detector (Perkin Elmer LC 235 C) and autosampler Perkin Elmer LC ISS series 200. The chromatographic system was controlled by the software package Turbochrom Version 4.1. plus and UV-spectrometric data were produced by TurboScan Version 2.0. A reverse phase Supelcosyl LC-8-DB, 250 x 4.6 mm I.D. (5 µm, particle size), protected by a guard column SupelguardTM LC-8-DB (2 cm) was used for separation. The mobile phase was consisted of 0.1 % (V/V) triethylamine in water with pH=3.5 and acetonitrile (63:37, V/V).
In order to achieve a good shape and location of diazepam peaks and the internal standard in the corresponding chromatograms, a series of parameters including composition and pH of mobile phase, column packing, flow rate and detection wavelength were tested. The final choice of the stationary phase giving satisfying resolution and run time was LC-8-DB. Triethylamine solution was prepared by adding 100 µL triethylamine in 100 mL H2O with pH adjusted to 3.5 with concentrated o-phosphoric acid. The mobile phase was filtered and degassed with helium. Chromatographic separations were performed at 37° C, with mobile phase flow rate of 1.3 mL/min and ultraviolet detection at 240 nm. The injection volume was 100 µL.
In order to achieve a good shape and location of diazepam peaks and the internal standard in the corresponding chromatograms, a series of parameters including composition and pH of mobile phase, column packing, flow rate and detection wavelength were tested. The final choice of the stationary phase giving satisfying resolution and run time was LC-8-DB. Triethylamine solution was prepared by adding 100 µL triethylamine in 100 mL H2O with pH adjusted to 3.5 with concentrated o-phosphoric acid. The mobile phase was filtered and degassed with helium. Chromatographic separations were performed at 37° C, with mobile phase flow rate of 1.3 mL/min and ultraviolet detection at 240 nm. The injection volume was 100 µL.
3
Quantitative Analysis of Testosterone
4
Quantitation of Testosterone by LC-MS/MS
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Quantitation of testosterone was performed in selected reaction monitoring (SRM) mode. Mass transitions and optimized MS/MS parameters are given in Table S5 . Analyst® software v1.4.1 (AB SCIEX) was used for SRM, peak integration, and analyte quantitation. Standard curves were prepared for testosterone in both tissue culture media and water in the range 0–20 ng/ml and the limit of detection (LOD) and lower limit of quantitation (LLOQ) were established in both matrices. The concentration of testosterone was measured in FM, SDM and APSCE media. Peak areas for these samples were quantified against the external calibration curves of testosterone (Table S6 ). Reverse phase chromatographic separation of testosterone was achieved using a Perkin Elmer Series 200 LC (Beaconsfield, UK) equipped with a Luna C8(2) column (3 μm; 20 × 4 mm i.d.) and SecurityGuard C18 column (4 × 3 mm) (Phenomenex, UK) maintained at 30 °C and a flow rate of 0.5 ml min−1 using the conditions in Table S7 . An API4000 triple quadrupole LC/MS/MS (Applied Biosystems, USA) was used for analysis with electrospray ionization (ESI) performed in positive ion mode using nitrogen gas with source parameters found in Table S8 .
5
Determination of Tellurium Speciation
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The total tellurium content was determined using the Elan 6100 DRC-e ICP-MS spectrometer (Perkin Elmer, Waltham, MA, USA). The ICP-MS apparatus was equipped with a standard ICP quartz torch, cross-flow nebulizer, and nickel cones. Table 1 presents operating parameters of the spectrometer. To separate the Te(IV) and Te(VI) species, a speciation apparatus set was applied. It consisted of an HPLC chromatograph (Perkin Elmer, USA) equipped with a Series 200LC Peltier oven, Series 200LC autosampler, and Series 200LC gradient pump. The sample from chromatographic column was introduced to ICP-MS by tubing system, automatic diverter and peristaltic pump. The diverter operates as an automatic switching valve to divert undesired portions of the eluate from the HPLC system to waste before the sample enters the ICP-MS. Soil samples were digested in a microwave oven (Microwave 3000, Anton Paar, Austria). Soil extractions were carried out using an ultrasonic cleaner (Sonic 5, Polsonic, Warszawa, Poland), and then the samples were centrifuged using a Beckman Coulter Avanti JXN-26 centrifuge (20,000 rpm, JA-25.50 Fixed-Angle Aluminum Rotor type).
6
Ascorbic Acid and β-Carotene Quantification
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For estimation of ascorbic acid content (ASA), 1 g of frozen fruit tissues was homogenised in 5 mL of ice-cold 6 % m-phosphoric acid (pH 2.8) containing 1 mM EDTA (Gossett and al., 1994) . The homogenate was centrifuged at 20,000 × g for 15 min at 4 °C. The supernatant was filtered through a 30-µm syringe filter, and 50 µL of the filtrate was analyzed using a HPLC system (PerkinElmer series 200LC and UV/VIS detector 200 LC, USA) equipped with a 5-µm column (Spheri-5 RP-18; 220 × 4.6 mm; Brownlee) and UV detection at 245 nm with 1.0 mL/min water (pH 2.2) as the mobile phase, run isocratically (Gahler and al., 2003) ß-carotene content β-carotene was extracted by grinding fruit tissues in a solution of 100% acetone containing CaCO3 (Jung, 2004) . The extracts were centrifuged at 16,000 × g for 10 min, and 20 µL of the resulting supernatants were used for HPLC analysis, as described by Gilmore and Yamamoto (1991) using the previously mentioned HPLC system. Solvent A (acetonitrile, methanol, Tris-HCl buffer 0.1 M, pH 8.0, 72:8:3) was run isocratically from 0 to 4 min followed by a 2.5 min linear gradient to 100% solvent B (methanol, hexane, 4:1) at a flow rate of 2 mL/min. The detector was set at 440 nm for the integration of peak areas after calibration with the external standard.
7
Ascorbate Quantification in Plant Tissues
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Ascorbate fractions were estimated by homogenizing of 1 g of frozen plant tissues and 5 mL of ice-cold 6% m-phosphoric acid (pH 2.8) containing 1mM EDTA (Gossett et al., 1994) . The homogenate was centrifuged at 20,000 × g for 15min at 4°C. The supernatant was filtered through a 30-μm syringe filter, and 50μL of the filtrate was analyzed using an HPLC system (PerkinElmer series 200 LC and UV/VIS detector 200 LC, USA) equipped with a 5-μm column (Spheri-5 RP-18; 220 × 4.6mm; Brownlee). The solvent used was H 2 O (pH 2.2 using H 3 PO 4 ), which was run isocratically with a flow rate of 0.75mL/min (Gahler et al., 2003) . The detector was set at 260nm for the integration of peak areas after calibration with the external standard ASA and the results were expressed as µg g -1 FM.
8
HPLC Analysis of NADP+ and NADPH
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For HPLC analysis using PerkinElmer series 200 LC and UV/VIS detector 200 LC, USA system set with a 5-μm column (Spheri-5 RP-18; 220×4.6 mm; Brownlee), samples (0.3 g) were extracted either by acid (0.6 M perchloric acid) for nicotinamide adenine dinucleotide phosphate (NADP + ) measurement or alkaline (0.5 M potassium hydroxide) extraction for measureement of NADPH. Centrifugation was performed for the extract at 10,000 × g at 4 °C for 10 min followed by neutralization by either 0.5 M KOH or 1 M KH 2 PO 4 and centrifugation again at 10,000xg at 4 °C for 10 min. Twenty μl of the supernatants were employed as declared by Caruso et al., (2004) .
9
HPLC-Based Compound Analysis Protocol
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The analysis was performed as described previously (1) (link). Briefly, an HPLC-System of Perkin-Elmer (Arcade, USA) was used. The equipment consisted of an autosampler (ISS 200), diode array detector (Diode Array LC 235C), quaternary pump (Series 200 LC, Perkin-Elmer), column thermostat (LC Oven 101 CC5904762), gas removal equipment (Series 200 online degasser), sample collector (FC 203B; Gilson, USA), and interface (600 Link, Perkin-Elmer). Parameters of the analytical column: Supelcosil RP C18 250 × 4.6 mm, 5 m (Supelco, USA). Mobile phase A consisted of 99.95 % acetonitrile and 0.05 % trifluoroacetic acid (TFA). Mobile phase C consisted of 99.95 % water and 0.05 % TFA.
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