A kind of uplc method for measuring the fingerprint spectrum of Andrographis paniculata medicinal material and four kinds of diterpene lactones
A technology of andrographolide and fingerprint spectrum is applied in the direction of measurement device, material separation, analysis of materials, etc., which can solve the problems of complicated operation and poor precision.
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Embodiment 1
[0145] Acquity UPLC (Waters, USA, including quaternary high-pressure gradient pump, vacuum degasser, autosampler, column oven, PDA detector, Empower3 chromatography workstation); KQ-500DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); XS105 1 / 100,000 analytical balance (Mettler Toledo, Switzerland); MilliporeSimplicity ultrapure water system.
[0146] Acetonitrile, methanol (chromatographic grade, Merck); formic acid (chromatographic grade, Tianjin Guangfu Fine Chemical Institute); Testing Institute; neoandrographolide reference substance (batch number: AD728N, content 98.18%) and deoxyandrographolide reference substance (batch number: AD159D, content 99.18%) were purchased from Tianjin Yifang Technology Company.
[0147] Andrographis paniculata (Burm.F.) Nees was identified as Andrographis paniculata (Burm.F.) Nees by Professor Wu Guixin of Shanghai University of Traditional Chinese Medicine, and crushed into Andrographis paniculata (Burm.F.) Nees before th...
Embodiment 2
[0165] Step 1: Preparation of the test solution
[0166] Get Andrographis paniculata medicinal material powder 500mg, accurately weighed. Add 25mL of 80% methanol-water solution for ultrasonic extraction for 20min. After ultrasonication, place it at room temperature, use 80% methanol-water to make up the volume according to the volume, filter through a 0.22μm microporous membrane, and take the subsequent filtrate to obtain the final product.
[0167] Step 2: Chromatographic Conditions
[0168] Adopt Acquity UPLC BEH C18 chromatographic column (1.7μm, 2.1×100mm); gradient elution, the elution procedure is as follows: 0~2min, 85%→77%B; 2~3min, 77%B; 3~5min, 77%B →76%B; 5~7min, 76%→75%B; 7~7.5min, 75%→69%B; 7.5~8min, 69%B; 8~9min, 69%→65%B; 9~11min , 65%→60%B; 11~12min, 60%→58%B; 12~13min, 58%B; 13~15min, 58%→35%B; 15~17min, 35%→20%B; 17 ~19min, 20%→85%B; 19~21min, 85%B. Mobile phase A is 0.3% formic acid-water solution, mobile phase B is methanol: acetonitrile = 1:9; injecti...
Embodiment 3
[0174] Step 1: Preparation of the test solution
[0175] Get Andrographis paniculata medicinal material powder 500mg, accurately weighed. Add 10mL of 60% methanol-water solution for ultrasonic extraction for 40min. After ultrasonication, place it at room temperature, use 60% methanol-water to adjust the volume, filter through a 0.22μm microporous membrane, and take the subsequent filtrate to obtain the final product.
[0176] Step 2: Chromatographic Conditions
[0177] Adopt Acquity UPLC BEH C18 column (1.7μm, 2.1×100mm); gradient elution, elution procedure is as follows: 0~2min, 88%→80%B; 2~3min, 80%B; 3~5min, 80%B →79%B; 5~7min, 79%→78%B; 7~7.5min, 78%→72%B; 7.5~8min, 72%B; 8~9min, 72%→68B; 9~11min, 68% →63B; 11~12min, 63→61B; 12~13min, 61B; 13~15min, 61→38%B; 15~17min, 38%→23%B; 17~19min, 23%→88%B; 19 ~21 min, 88%B. Mobile phase A is 0.2% formic acid aqueous solution, mobile phase B is methanol: acetonitrile = 2:8 mixed solution; injection volume is 1 μL; column tempera...
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