Chromolith rp 18e

Manufactured by Merck Group

Sourced in Germany, United States

Chromolith RP-18e is a high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a wide range of organic compounds. The column is packed with a silica-based stationary phase that provides efficient chromatographic separation.

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Lab products found in correlation

1100 series, by Merck Group (5 mentions) 1200 series, by Merck Group (5 mentions) Diode array detector, by Agilent Technologies (3 mentions) 6130 series, by Merck Group (3 mentions) Inova 300, by Agilent Technologies (3 mentions) Agilent 1100 series, by Agilent Technologies (1 mentions) Gamma detector, by Bioscan (1 mentions) Maldi ms daltonics microflex, by Bruker (1 mentions) Labsolutions software version 5.75 sp2, by Shimadzu (1 mentions) Md 4010, by Jasco (1 mentions) As 4050, by Jasco (1 mentions) Pu 4180, by Jasco (1 mentions) Cbm 20a, by Shimadzu (1 mentions) Chromolith semiprep rp 18e, by Merck Group (1 mentions) Tlc silica gel 60 f254, by Merck Group (1 mentions) Sil htc autosampler, by Shimadzu (1 mentions) Cto 10a, by Shimadzu (1 mentions) Spd m20a, by Shimadzu (1 mentions) Lcms 2020, by Shimadzu (1 mentions) Dgu 20a3, by Shimadzu (1 mentions)

Spelling variants of this product

RP-18e (13 citations) Chromolith® Performance RP-18e (11 citations) Chromolith® Performance RP-18e 100–4.6 mm (9 citations) Chromolith RP-18e column (6 citations) Chromolith FastGradient RP-18e HPLC column (3 citations) Chromolith® FastGradient RP-18e column (3 citations) Chromolith Rp-18e 100 x 4.6 mm column (2 citations) Chromolith Caprod RP-18e HR capillary column (2 citations) Chromolith Performance RP-18e 100-2 mm column (2 citations) Chromolith® HighResolution RP-18e column (1 citations)

21 protocols using chromolith rp 18e

Synthesis and Characterization of PSMA PLT-TM

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Commercially available analytical grade chemicals were used and not further purified. The chemical suppliers were: Sigma-Aldrich (Taufkirchen, Germany) and Merck (Darmstadt, Germany), unless otherwise indicated. Protected amino acids and resins were supplied from Novabiochem (Merck, Darmstadt, Germany) and IRIS Biotech (Marktredwitz, Germany).
The following RP-HPLC systems were used for analysis and purification: Agilent 1100 Series, equipped with a multi-wavelength-detector and Latek P402 (Latek Labortechnik-Geraete, Eppelheim, Germany) equipped with a HITACHI variable UV detector (absorbance measurement at 214 and 254 nm) and a gamma detector (Bioscan, Washington, USA). Both RP-HPLC systems were equipped with either an analytical Chromolith RP-18e (4.6 × 100 mm; Merck, Darmstadt, Germany) for analysis or a semi-preparative column Chromolith RP-18e (10 × 100 mm; Merck, Darmstadt, Germany) for purifications.
The MALDI-MS Daltonics Microflex (Bruker Daltonics, Bremen, Germany) system was used for mass spectrometry. Full-scan single mass spectra were obtained by scanning m/z = 400–4000.
Synthesis of PSMA PLT-TM is summarized in Figure 2 and described in detail in the following chapters:

Arndt C., Feldmann A., Koristka S., Schäfer M., Bergmann R., Mitwasi N., Berndt N., Bachmann D., Kegler A., Schmitz M., Puentes-Cala E., Soto J.A., Ehninger G., Pietzsch J., Liolios C., Wunderlich G., Kotzerke J., Kopka K, & Bachmann M. (2019). A theranostic PSMA ligand for PET imaging and retargeting of T cells expressing the universal chimeric antigen receptor UniCAR. Oncoimmunology, 8(11), 1659095.

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LC-MS Analysis of 2,3-Dehydroflavonolignans

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2,3-Dehydroflavonolignans from silymarin were analyzed by an LC-MS method on monolithic Chromolith RP-18e (100 × 3 mm) column (Merck, Darmstadt, Germany), Chromolith RP-18e (5 × 4.6 mm) pre-column (Merck, Darmstadt, Germany). Binary gradient elution was performed using mobile phase A = 5% acetonitrile, 0.1% formic acid, B = 80% acetonitrile, 0.1% formic acid, and gradient: 0 min 20% B, 5 min 90% B, 6 min 90% B, 8–10 min 20% B; flow rate 0.4 mL/min, 25 °C, MS detection. The MS parameters were as follows: Negative mode; ESI interface voltage, 4.5 kV; detector voltage, 1.15 kV; nebulizing gas flow 1.5 mL/min; drying gas flow 15 mL/min; heat block temperature 200 °C; DL temperature 250 °C; SCAN mode 450–650 m/z. Spectra were extracted in the 479.0–479.1 m/z range ([M-H]⁻ ions of 2,3-dehydroflavonolignans) using LabSolutions software version 5.75 SP2 (Shimadzu, Kyoto, Japan). The injection volume was 1 μL.

Petrásková L., Káňová K., Biedermann D., Křen V, & Valentová K. (2020). Simple and Rapid HPLC Separation and Quantification of Flavonoid, Flavonolignans, and 2,3-Dehydroflavonolignans in Silymarin. Foods, 9(2), 116.

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Isocratic Separation of Silymarin

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The original isocratic method for the separation of silymarin components was performed under isocratic conditions on monolithic Chromolith RP-18e (100 × 3 mm) column with monolithic Chromolith RP-18e (5 × 4.6 mm) pre-column (both Merck, Darmstadt, Germany), mobile phase: 2% acetonitrile, 37% methanol, 0.1% formic acid, flow rate 1.1 mL/min, t = 25 °C. The PDA data were acquired in the 200–400 nm range, sampling 40 Hz, time constant 0.025 s, and signals at 285 nm were extracted. The injection volume was 2 μL.

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Cytotoxicity of IQB-CBI Dimers

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Example 3

Cytotoxicity of D801, D803, D05, D807, D809, D811, D813, D815 and D817 were tested against AML2 and HL60 cell lines. Results are shown in Table 1. Cell killing assay was performed after 3-day incubation with various IQB-CBI dimers.

TABLE 1

IC50

pg/mLD211D801D803D805D807D809D811D813D815D817

AML20.1916.214.652.21322860.130.280.070.13

HL600.4744.540.6142.655718500.290.720.120.19

General Methods:

¹H NMR spectra were recorded on a Varian Inova 300 or 500 MHz NMR instrument. Chromatographic purities were determined on an Agilent 1200 Series, 1100 Series or 6130 Series LC/MS system using a Merck Chromolith RP-18e analytical HPLC column (monolithic, 50×2 mm) and the following analytical HPLC method: injection volume 5 μL; flow rate 1 mL/min; 5→95% acetonitrile in water with 0.05% AcOH over 5 mins; Agilent diode array detector at λ=254, 220 or 195 nm; room temperature.

US10350218B2. Isoquinolidinobenzodiazepine (IQB)-1(chloromethyl)-2,3-dihydro-1H-benzo[e]indole (CBI) dimers (2019-07-16). Cellerant Therapeutics, Inc. [US]. Inventors: Jagath R. Junutula [US], Sean W. Smith [US], Dmitry Borkin [US], Sylvia Degrado [US], Sanjeevani Ghone [US].

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HPLC Analysis of Nicotinic Acid

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NIA analysis was conducted using an HPLC equipped with a reversed-phase monolithic column (Chromolith^® RP-18e, 100 mm × 4.6 mm i.d., Merck) connected to a Jasco (Easton, PA, USA) HPLC system (pump PU-4180, autosampler AS-4050 and LC-Net II/ADC controller) coupled to a PDA detector (Jasco MD-4010, start wavelength = 200 nm, End Wavelength = 400 nm). The data processing was performed by ChromNAV 2.0 HPLC software (Easton, PA, USA). The chromatogram was acquired at a wavelength of 263 nm, and the retention time of NIA was at 4.8 min. The mobile phase was a mixture of water and methanol (80:20, v/v) and eluted at a flow rate of 1.0 mL/min. The injection volume was set to 20 μL with the column temperature at 30 °C. Prior to use, the mobile phase was degassed in an ultrasonic bath for 30 min. Validation of HPLC method was performed by evaluating linearity, range accuracy, precision, lower limit of quantification (LOQ), and upper limit of detection (LOD) according to established guidelines [43 (link)]. The calculated LOD was 0.18 µg/mL and the LOQ was 0.53 µg/mL.

Basto R., Andrade R., Nunes C., Lima S.A, & Reis S. (2021). Topical Delivery of Niacinamide to Skin Using Hybrid Nanogels Enhances Photoprotection Effect. Pharmaceutics, 13(11), 1968.

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Isolation of 2,3-Dehydroflavonolignans

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The LC system used for isolation of 2,3-dehydrosilybin and 2,3-dehydrosilychristin consists of an LC-8A preparative HPLC pump, SPD-20A dual-wavelength UV detector, FRC-10A fraction collector, and CBM-20A bus module controlled by the LC solution 1.24 SP1 software (Shimadzu, Kyoto, Japan).
Semi-preparative chromatography of the fraction enriched in 2,3-dehydroflavonolignans (see Section 2.1 and Figure A1 in Appendix B) was conducted using monolithic Chromolith SemiPrep RP-18e (100 × 10 mm, Merck, Darmstadt, Germany) column with monolithic Chromolith RP-18e (5 × 4.6 mm) pre-column in mobile phases containing either 50% methanol, 0.1% formic acid, 5 mL/min flow rate or 45% methanol, 5% acetonitrile, 0.1% formic acid, 5 mL/min flow rate. Both mobile phases produced similar results. Alternatively, ASAHIPAK GS-10G 7B (300 × 25 mm, 20 μm, Shodex, Munich, Germany) column was used with mobile phase 50% methanol, 0.1% formic acid at 5 mL/min flow rate. The injection volume was 300 µL (37 mg/mL) in all cases.

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Radiolabeling of Gallium-68, Scandium-44, and Lutetium-177

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For radiochemical evaluation, gallium-68 was eluted from a ⁶⁸Ge/⁶⁸Ga-generator (ITG Graching, Munich, Germany) and purified manually with ethanol-based post-processing to separate iron, zinc, and germanium impurities [44 (link)].
Radiolabeling was performed in 0.4 mL 1 M ammonium acetate buffer at pH 5.5. Reactions were carried out with different amounts of precursor (5, 10, 15, 60 nmol) and at different temperatures (RT, 50 °C, and 70 °C) with 30–50 MBq gallium-68. The pH was controlled at the start and after the labeling. For reaction control, radio-TLC (TLC Silica gel 60 F₂₅₄ Merck) and citrate buffer pH 4 as mobile phase and radio-HPLC using an analytical HPLC 7000 series Hitachi LaChrom (Column: Merck Chromolith^® RP-18e, linear gradient of 5–95% MeCN (+0.1% TFA)/95–5% Water (+0.1% TFA) in 10 min). TLCs were measured in a TLC imager CR-35 Bio Test-Imager (Elysia-Raytest, Angleur, Belgium) with the analysis software AIDA (Elysia-Raytest, Angleur, Belgium).
Radiolabeling of AAZTA⁵.SA.KuE with scandium-44 and lutetium-177 was performed according to the literature [17 (link)].

Lahnif H., Grus T., Pektor S., Greifenstein L., Schreckenberger M, & Rösch F. (2021). Hybrid Chelator-Based PSMA Radiopharmaceuticals: Translational Approach. Molecules, 26(21), 6332.

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NMR Spectroscopy and HPLC Analysis

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Example 4

General Methods:

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Analytical HPLC Method for Chromatographic Purity

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Example 29

[Figure (not displayed)]

Analytical methods: Chromatographic purities were determined on an Agilent 1200 Series, 1100 Series or 6130 Series LC/MS system using a Merck Chromolith RP-18e analytical HPLC column (monolithic, 50×2 mm) and the following analytical HPLC method: injection volume 5 μL; flow rate 1 mL/min; 5-95% acetonitrile in water with 0.05% AcOH (Method A) or 0.05% TFA (Method B) over 5 mins; Agilent diode array detector at 1=254, 220 or 195 nm; room temperature.

US11634508B2. Peptide conjugates of cytotoxins as therapeutics (2023-04-25). Cybrexa 2, Inc. [US]. Inventors: Daniel Richard Marshall [US], Johanna Marie Csengery [US], Robert John Maguire [US], Robert A. Volkmann [US].

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Quantification of AML, VAL and HCTZ by HPLC-MS/MS

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A Shimadzu LC-VP HPLC system (Kyoto, Japan) consisting of LC-10ADVP pump, SIL-HTc autosampler, CTO 10 ASvp column oven and a DGU-14A degasser was used for setting the reverse-phase liquid chromatographic conditions. The separation of AML, VAL and HCTZ was achieved on a Chromolith RP_18e (100 mm × 4.6 mm) analytical column from Merck KGaA (Darmstadt, Germany) and maintained at 35 °C in a column oven. For isocratic separation, the mobile phase consisted of acetonitrile and 2 mM ammonium formate, pH 4.0 adjusted with formic acid (90:10, v/v). Ionization and detection of AML, VAL, HCTZ and ISs was carried out on a triple quadrupole mass spectrometer, MDS SCIEX API-4000 (Toronto, Canada), equipped with turbo ion spray interface and operated in positive ionization mode for AML and VAL and negative mode for HCTZ. The set chromatographic conditions and mass parameters are described in Supplementary material.

Shah J.V., Parekh J.M., Shah P.A., Shah P.V., Sanyal M, & Shrivastav P.S. (2017). Application of an LC–MS/MS method for the analysis of amlodipine, valsartan and hydrochlorothiazide in polypill for a bioequivalence study. Journal of Pharmaceutical Analysis, 7(5), 309-316.

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