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28
2026
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08
Analysis of Flumazenil, referring to the Chinese Pharmacopoeia (2025 edition), Volume II
Flumazenil is 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylate ethyl ester.
Flumazenil C15H14FN3O3
303.29 CAS No:78755-81-4
01
Basic Info
This product is a white or off-white crystalline powder; it has no smell. It dissolves easily in glacial acetic acid, slightly in methanol, and is almost insoluble in water.
1．Execution Standard
'Chinese Pharmacopoeia' (2025 edition), Volume II, Flumazenil.
2．Test sample
Flumazenil
02
Solution Preparation
1. Test solution: Take this product, dissolve it in the mobile phase, and dilute it to make a solution containing 1.0 mg per 1 mL.。
2. Reference solution: Accurately measure an appropriate amount of the test solution, then dilute it with the mobile phase to prepare a solution containing 10 μg per 1 mL.
3. Sensitivity solution: Accurately measure an appropriate amount of the reference solution and dilute it with the mobile phase to make a solution containing about 0.5 μg per 1 mL.
4. Blank solution: Use the mobile phase as the blank solution.
03
System Applicability Requirements
The theoretical plate number, calculated based on the Flumazenil peak, should be no less than 3000.
04
CP C18 MGII
1. HPLC Chromatography Condition
Column：CP C18 MGII 5 μm 4.6×250 mm；
Column temperature：35 ℃；
Mobile phase：A dilute phosphoric acid solution (take 1000 mL of water and adjust the pH to 2.0 with phosphoric acid) - methanol - tetrahydrofuran (80:13:7) as the mobile phase;
flow rate：1.0 mL/min；
Detection wavelength：230 nm；
Injection Volumn：20 μL，Record the chromatogram up to three times the retention time of the main component peak.
2．Reference solution chromatogram
System Applicability Data Sheet
3．Chromatogram of related substances in the test solution
4．Sensitivity Solution Chromatogram
Signal-to-noise ratio S/N: 36.1
5．Blank Solution Chromatography
05
DP ODS-P
1. HPLC Chromatography Condition
Column：DP ODS-P, 5 μm 4.6×250 mm；
Column Tremperature：35 ℃；
Mobile Phase：Use a dilute phosphoric acid solution (take 1000 mL of water and adjust the pH to 2.0 with phosphoric acid) - methanol - tetrahydrofuran (80:13:7) as the mobile phase;
Flow rate：1.0 mL/min；
Detective wavelength：230 nm；
Injection Volumn：20 μL，Record the chromatogram up to three times the retention time of the main component peak.
2．Chromatogram of the reference solution
System Applicability Data Sheet
3．Chromatogram of related substances in the test sample solution
4．Sensitivity Solution Chromatogram
Signal-to-noise ratio S/N: 31.5
5．Blank solution chromatogram
06
Summary
In summary, referring to the analysis method for flumazenil in Volume II of the Chinese Pharmacopoeia (2025 edition), HPLC analysis was performed using CAPCELL PAK C18 MGII, 5 μm 4.6×250 mm and DAISOPAK SP-100-5-ODS-P, 4.6×250 mm columns, and the results all met the pharmacopoeia requirements.
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