CLZ assay was performed by HPLC (Merck Hitachi Elite LaChrom apparatus, Darmstadt, Germany) endowed with a L-2400 UV-vis detector and a L-2130 isocratic pump. An acetonitrile/water 30:70 v/v mixture was used as mobile phase. An Agilent Zorbax CN column (150 × 4.6 mm, 5 µm particle size) was the stationary phase. UV detection was performed at 310 nm. The injection volume 20 µL, the flow rate 0.9 mL/min, the column temperature 40 ± 1.0 °C. Under these conditions, the CLZ retention time was 7.23 ± 0.01 min. The method was validated for linearity (r2 = 0.9985), limit of quantification (0.27 µg/mL) and limit of detection (0.08 µg/mL).
L 2130
Manufactured by Merck Group
Sourced in Germany, Japan
The L-2130 is a laboratory equipment product manufactured by Merck Group. It is designed for performing various scientific experiments and analyses in a controlled laboratory environment. The core function of the L-2130 is to provide a reliable and consistent platform for conducting research and testing procedures.
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Lab products found in correlation
L 2455, by Merck Group (3 mentions)
Elite lachrom apparatus, by Merck Group (2 mentions)
L 2400, by Merck Group (2 mentions)
L 2200, by Merck Group (2 mentions)
Hplc dad, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Jetstream 2 plus, by Knauer (1 mentions)
Nylon 66 membrane filter, by Cytiva (1 mentions)
Ezchrom elite software, by Merck Group (1 mentions)
Zorbax eclipse xdb c18, by Agilent Technologies (1 mentions)
Elitelachrom, by Merck Group (1 mentions)
6 protocols using l 2130
1
HPLC Method for Clozapine Quantification
2
HPLC Quantification of P293BL and P297
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Compound assay was performed by HPLC (Merck Hitachi Elite LaChrom apparatus, Darmstadt, Germany), equipped with a L-2400 UV–Vis detector and an L-2130 isocratic pump. The mobile phase A was water, and the mobile phase B was methanol. Gradient steps were programmed as follows: A–B 60%–40% at time 0, ramped to 70% B in 3 min, held at 70% for 2 min and returned to initial conditions during 3 min. A Hibar Purospher® RP-8e (150 × 4.6 mm, 5 µm pore size) was the stationary phase. UV detection was performed at 292 nm. The injection volume was 20 µL, the flow rate was 1.2 mL/min and the column temperature was 40 ± 1 °C. Under these conditions, the compounds retention time was 5.30 ± 0.01 min. The method was validated for linearity (r2 = 0.998; r2 = 0.997), limit of quantification (3.03 µg/mL; 4.50 µg/mL) and limit of detection (0.91 µg/mL; 1.35 µg/mL) for P293BL and P297, respectively.
3
Quantitative HPLC Analysis of Chlorogenic Acids
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The instrumental analysis of CQAs was performed using HPLC-DAD, Merck Hitachi Elite La Chromatograph (Tokyo, Japan) equipped with a quaternary system of pumping (L-2130) and L-2455 UV/vis spectrophotometry diode array detector. Separation was achieved using LiChroCART RP-18 endcapped (250 × 4 mm, 5 μm) column, attached to a guard column (4 × 4 mm, 5 μm) of the same kind.
Quantitative analysis of chlorogenic acids was performed based on the method described previously by Tfouni et al. [15 (link)] with slight modifications. The mobile phase was constituted eluent A: 10 mM citric acid aqueous solution (pH of 2.4) and eluent B: acetonitrile. The gradient was programmed as follows: from 0 to 30 min 8% of B, 30 to 35 min increase to 80% of B, 35 to 40 min 80% of B, 40 to 45 min decrease to 8% of B, and 45 to 50 min 8% of B. Injected volume was 10 μL and the flow rate of analysis was 1 mL/min. Detection of CQAs was carried out at 325 nm. Identification of the target compounds was confirmed by retention time and spectrum comparison with standard solutions.
Quantitative analysis of chlorogenic acids was performed based on the method described previously by Tfouni et al. [15 (link)] with slight modifications. The mobile phase was constituted eluent A: 10 mM citric acid aqueous solution (pH of 2.4) and eluent B: acetonitrile. The gradient was programmed as follows: from 0 to 30 min 8% of B, 30 to 35 min increase to 80% of B, 35 to 40 min 80% of B, 40 to 45 min decrease to 8% of B, and 45 to 50 min 8% of B. Injected volume was 10 μL and the flow rate of analysis was 1 mL/min. Detection of CQAs was carried out at 325 nm. Identification of the target compounds was confirmed by retention time and spectrum comparison with standard solutions.
4
HPLC-MS Analysis of Organic Compounds
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All chemical solvents and standards were of analytical grade. Acetonitrile was delivered from Merck (Darmstadt, Germany) and formic acid from POCh (Katowice, Poland). HPLC ELITE LaChrom VWR (Merck) instrumentation consisted of the following components: a pump series L-2130 in gradient mode, an autosampler series L-2200 with a 100 μL injection loop, Column Oven series L-2350, and the diode array detector (DAD) series L-2455. The chromatographic separation was performed in the gradient mode with a Discovery RP 18 column (250 × 4.6 mm I.D., 5 μm particle size) from Sigma-Aldrich (Supelco).
The mobile stage consisted of the phase A which was 0.1% v/v formic acid (HCOOH) in water and the phase B which was Acetonitrile (0.050% TFA in Acetonitrile/water 80 : 20 v/v). The flow rate increased from 0.6 mL/min to 1.2 mL/min for 11minutes and then decreased to 0.6 mL/min. The gradient was programmed as follows: 6% B for 11 min, then increased to 14% B for 3 min, and then decreased to 6% B. Temperature of the column was equal to 15°C. The applied gradient was linear from 0 to 55% in 11 min, at a flow rate of 1.2 mL/min.
The DAD was set at a wavelength of 274 nm. Mass spectra were collected every 3 ms in the positive ion mode. MS spray voltage was 4.50 kV and the capillary temperature was 250°C.
The mobile stage consisted of the phase A which was 0.1% v/v formic acid (HCOOH) in water and the phase B which was Acetonitrile (0.050% TFA in Acetonitrile/water 80 : 20 v/v). The flow rate increased from 0.6 mL/min to 1.2 mL/min for 11minutes and then decreased to 0.6 mL/min. The gradient was programmed as follows: 6% B for 11 min, then increased to 14% B for 3 min, and then decreased to 6% B. Temperature of the column was equal to 15°C. The applied gradient was linear from 0 to 55% in 11 min, at a flow rate of 1.2 mL/min.
The DAD was set at a wavelength of 274 nm. Mass spectra were collected every 3 ms in the positive ion mode. MS spray voltage was 4.50 kV and the capillary temperature was 250°C.
5
Isocratic HPLC Characterization of Retention Factors
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The retention factors were measured with a liquid chromatograph and an Elite LaChrom HPLC Merck-Hitachi (Merck, Darmstadt, Germany) with a DAD (Diode Array Detector L-2455) detector, pomp L-2130, and a manual sample injection valve equipped with a 20 µL loop and EZChrom Elite software (Merck, Darmstadt, Germany) system manager as a data processor. The column was a Zorbax Eclipse XDB-C18 (Agilent Technologies, Santa Clara, CA, USA); (150 mm × 4.6 mm I.D., 5 µm). The injection of blank mobile phase volumes produced visible detector fluctuations that were used as the hold-up volume. The column was thermostated at 20.0 °C ± 0.1 using a column thermostat Jetstream 2 Plus (100375, Knauer, Berlin, Germany). The elution was carried out in the isocratic mode by the mobile phase consisting of 80% methanol in water. The mobile phase was filtered through a Nylon 66 membrane filter (0.45 μm) Whatman (Maidstone, Kent, England) by the use of a filtration apparatus.
The retention data were recorded at a flow rate of 1 mL min−1 with online degassing using L-7612 solvent degasser at a wavelength chosen accordingly with the recorded spectra in the range of 200–600 nm. Typical injection volumes were 20 µL. HPLC measurements were performed in triplicate.
The retention data were recorded at a flow rate of 1 mL min−1 with online degassing using L-7612 solvent degasser at a wavelength chosen accordingly with the recorded spectra in the range of 200–600 nm. Typical injection volumes were 20 µL. HPLC measurements were performed in triplicate.
6
HPLC Analysis of Diterpene Compounds
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HPLC analysis was performed in a Merck Hitachi Elite LaChrom (Tokyo, Japan) system equipped with a quaternary pump (L-2130), an L-2200 autosampler and a L-2455 UV/vis spectrophotometry diode array detector. Separation was achieved using a Purospher STAR LichroCART RP 18 end-capped (250 × 4 mm, 5 μm) column attached to a guard column (4 × 4 mm, 5 μm) of the same kind. The detection wavelengths were 225 nm for cafestol and 290 nm for kahweol. EZChrom Elite 3.1.6 software was used for data acquisition and analysis.
Before chromatographic analysis, the dried extracts were made up to 2.5 mL (ECs) or 10 mL (R&G Arabica coffee) with acetonitrile. Twenty microliters of the reconstituted samples were injected after filtration (0.45 µm polytetrafluoroethylene membranes (PTFE), VWR, USA). Mobile phase was acetonitrile/water (55/45, v/v) with an isocratic flow rate of 0.8 mL/min [27] (link). Target compounds were identified by comparing spectra and retention times of reference standard solutions. Quantitative analysis was performed using external standard calibration curves by plotting the peak area vs. the corresponding concentrations.
Before chromatographic analysis, the dried extracts were made up to 2.5 mL (ECs) or 10 mL (R&G Arabica coffee) with acetonitrile. Twenty microliters of the reconstituted samples were injected after filtration (0.45 µm polytetrafluoroethylene membranes (PTFE), VWR, USA). Mobile phase was acetonitrile/water (55/45, v/v) with an isocratic flow rate of 0.8 mL/min [27] (link). Target compounds were identified by comparing spectra and retention times of reference standard solutions. Quantitative analysis was performed using external standard calibration curves by plotting the peak area vs. the corresponding concentrations.
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