All chemicals were purchased from commercial sources (MicroSource Discovery Systems, Aldrich Market Select, Sigma, Sequoia Sciences and Tocris Bioscience) and their identity confirmed by mass spectrometry in the Chemical Synthesis Core (Agilent 1200 LCMS).
Agilent 1200 lc ms
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1200 LC-MS is a liquid chromatography-mass spectrometry system. It is designed for the separation, identification, and quantification of chemical compounds. The system combines a high-performance liquid chromatograph with a mass spectrometer, enabling the analysis of complex samples with high sensitivity and specificity.
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5 protocols using agilent 1200 lc ms
1
Chemical Identification Protocol
2
Quantification of Paclitaxel Distribution in Glioma Mice
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Forty five glioma-bearing ICR mice were randomly divided into three groups and intravenously injected with Taxol®, NP-PTX and Pep-NP-PTX at the dose of 8 mg/kg PTX, respectively. At different time points (0.5, 1 and 4 h, n = 5 at each time point) after injection, brain and glioma of the mice were collected. The tissues were homogenized in 0.9% sodium chloride solution with 1% TritonX-100 after the weight measurement. The supernatant was obtained after centrifugation.
For the determination of PTX, 100 μL supernatant of tissue homogenates (or 100 μL plasma) were mixed with 100 μL methanol containing 100 ng/mL docetaxel (internal standard). Then 1 mL ether was added to extract the PTX and docetaxel. After dried the organic phase under N2, the residue was dissolved with 200 μL 80% methanol solution and analyzed by HPLC-mass spectrometry (Agilent 1200 LC-MS, USA).
For the determination of PTX, 100 μL supernatant of tissue homogenates (or 100 μL plasma) were mixed with 100 μL methanol containing 100 ng/mL docetaxel (internal standard). Then 1 mL ether was added to extract the PTX and docetaxel. After dried the organic phase under N2, the residue was dissolved with 200 μL 80% methanol solution and analyzed by HPLC-mass spectrometry (Agilent 1200 LC-MS, USA).
3
Characterization of Organic Compounds by NMR and MS
4
Synthesis and Characterization of Organic Compounds
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All commercial reagents and solvents were purchased and used without additional purification. The progression of the reaction was detected using thin layer chromatography (TLC), it was performed on silica gel 60 F254 plates (Jiangyou, Yantai). Column chromatography was run on silica gel (200–300 mesh) from Qingdao Ocean Chemicals (Qingdao, Shandong, China). The melting points of all compounds were determined with a Büchi Melting Point B-540 apparatus (Büchi Labortechnik, Flawil, Switzerland) and were uncorrected. The mass spectra (MS) were determined in the electrospray ionization (ESI) mode on an Agilent 1200 LC-MS (Agilent, Palo Alto, CA, USA). Nuclear magnetic resonance (1H-NMR and 13C-NMR) spectra were recorded on a Bruker 500 MHz NMR spectrometer using tetramethylsilane (TMS) as an internal standard. The chemical shifts were reported in parts per million (ppm) and the coupling constants (J) were reported in Hertz (Hz). Peak multiplicities were expressed as follows: s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; dt, doublet of triplets; td, triplet of doublets; ddd, doublet of doublet of doublets; m, multiplet; and br, broad.
5
Optimized Synthesis and Characterization of Novel Compounds
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Unless otherwise noted, all reagents and solvents were obtained from commercially available sources and were used without purification. TLC analysis was performed on GF254 silica gel plates (Jiangyou, Yantai). Column chromatography was carried out with silica gel (200–300 mesh) from Qingdao Haiyang Chemicals (Qingdao, Shandong, China). Mass spectrometry was performed using ESI mode on an Agilent 1200 LC-MS (Agilent, Palo Alto, CA, USA). High-resolution accurate mass determinations (HRMS) were recorded on a Bruker Micromass Time of Flight mass spectrometer equipped with electrospray ionisation (ESI). Nuclear magnetic resonance (1H-NMR and 13C-NMR) spectra were recorded on a Bruker 400 MHz and 500 MHz NMR spectrometer with TMS as an internal standard. The chemical shifts were reported in parts per million (ppm), the coupling constants (J) were expressed in hertz (Hz). Peak multiplicities were described as singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m) and broad (br).
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