Silica gel 60 gf254

Electrospray ion trap mass spectrometry (ESI-MS) was carried out with a Bruker ESI-TRAP Esquire 6000 plus mass spectrometry instrument. Nuclear magnetic resonance spectra (NMR) were recorded on a Bruker Avance III 500 MHz instrument in DMSO-d6 using tetramethylsilane (TMS) as the internal standard. Analytical thin-layer chromatography (TLC) was performed with silica gel plates using silica gel 60 GF₂₅₄ (Qingdao Haiyang Chemical Co., Ltd.). Sephadex LH-20 was purchased from GE Healthcare Bio-science AB (Sweden).

All solvents and substances used were commercially purchased and not further purified. The reactions were monitored by thin-layer chromatography (TLC) with silica gel plates using silica gel 60 GF₂₅₄ (Qingdao Haiyang Chemical Co., Ltd., Qingdao, China). ¹H−NMR (400 MHz) and ¹³C−NMR (100 MHz) were measured by an AM−500 FT−NMR spectrometer (Bruker Corporation, Fällanden, Switzerland) with CDCl₃, acetone-d₆ or DMSO-d₆ as the solvent and TMS as the internal standard. MS was recorded under ESI conditions using LCQ Fleet instruments (Thermo Fisher, Waltham, MA, USA). The yields of the reactions were measured before optimization.

Technical products of Isoprothiolane (IPT) and Phenazine-1-carboxylic acid (PCA) were provided by the College of Agriculture, Yangtze University. All chemicals and solvents were commercially obtained and used directly without further purification. Column chromatography was performed over pure silica gel 60 (200–300 mesh, Liang Chen Gui Yuan Co., Ltd., Anhui, China). Thin-layer chromatography (TLC) was performed on silica gel 60 GF254 (Qingdao Hai Yang Chemical Co., Ltd., Qingdao, China). The melting points of all target compounds were determined by using a WRR melting point apparatus (Shanghai Jingke Industrial Co., Ltd., Shanghai, China) and are uncorrected. ¹H and ¹³C NMR spectra were recorded in DMSO-d₆ or CDCl₃ as the solvent using an AVANCE III HD 400 MHz spectrometer (Bruker Co., Ltd., Fällanden, Switzerland). Tetramethylsilane was used as the internal standard. High-Resolution Mass Spectra (HRMS) were obtained using electrospray ionization (ESI) technique by collision-induced dissociation on a Thermo Scientific Q Exactive instrument (Thermo Fisher Scientific, Bremen, Germany). The samples were dissolved in HPLC methanol and filtered by a 0.2 μm membrane. Then, it was infused using a syringe pump with a flow rate of 10 μL/min, and analyzed in the positive mode within a mass range m/z 100–1500.

A commercial bactericide of Thiodiazole copper, commercial fungicides Thifluzamide and Carbendazim were supplied by the College of Agriculture, Yangtze University. All reagents and solvents used in the experiment were provided by professional suppliers and utilized without further purification unless otherwise indicated. ¹H NMR and ¹³C NMR of all target compounds were performed on a AVANCE DPX 400 spectrometer (Bruker Co., Ltd., Fällanden, Switzerland) using tetramethylsilane (TMS) and dimethyl sulfoxide-d₆ (DMSO-d₆) as an internal standard and solvent, respectively (2.50 ppm for ¹H and 39.52 ppm for ¹³C). High-resolution mass spectrometry (HRMS) data were acquired from Thermo Scientific Q Exactive (Thermo Fisher Scientific, Bremen, Germany). All reactions were indicated via thin-layer chromatography (TLC) utilizing silica gel 60 GF254 (Qingdao Hai Yang Chemical Co., Ltd., Qingdao, China). The melting point of the target compounds were measured by WRR melting point apparatus, provided by Shanghai Precision Scientific Instrument Co., Ltd., Shanghai, China.

All reagents and solvents were of reagent grade or purified according to standard methods before use. Analytical thin-layer chromatography and preparative thin-layer chromatography were performed with silica gel plates using silica gel 60 GF254 (Qingdao Haiyang Chemical Co., Ltd., Qingdao, Shandong Province, China). ¹H-NMR and ¹³C NMR spectra were recorded at 600 MHz on a Bruker Avance-600 superconducting nuclear magnetic resonance instrument using TMS as a reference (Bruker Company, Gemany). High-resolution mass spectrometry (HRMS) data were obtained on an Uplc1290-6540B Q-TOF LC/MS instrument (Agilent Co., Santa Clara, CA, USA). The key intermediate 1-substituted-β-carboline-3-carbohydrazide derivatives 1 were prepared according to the procedure reported previously [26 (link)].

All reagents and solvents were of reagent grade or were purified according to standard methods before use. Analytical thin-layer chromatography (TLC) was performed with silica gel plates using silica gel 60 GF₂₅₄ (Qingdao Haiyang Chemical Co., Ltd., Qingdao, China). Melting points were determined on an XT-4 digital melting point apparatus (Beijing Tech Instrument Co., Ltd., Beijing, China) and were uncorrected. Proton nuclear magnetic resonance spectra (¹H NMR) were recorded on a Bruker Avance DMX 400 MHz instrument (Bruker, Bremerhaven, Germany) in CDCl₃ or DMSO-d₆ using TMS (tetramethylsilane) as the internal standard. High-resolution mass spectrometry (HRMS) was carried out with a Xevo G2-SQTOF instrument (Waters, Milford, MA, USA).