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Analysis of some prohibited drugs in Chinese herb extracts by HPLC /MS
Abstract: [Objective] To establish a method to detect the prohibited drugs in Chinese herb extracts. [Methods] It includes the extraction of drugs in organic solutions and their detection by HPLC /MS. For those positive samples their molecular structurs tested through HPLC / MS/MS.[Results]The scanning method for the identification of 65 drugs were established through their individual retention time, their specialmass and buy use of HPLC /MS/MS. [Conclusion] This paper introduces an analytical system to recognize and confirm p Robinated drugs in Chinese herb extracts bymass spectrum. It provides a necessary technological support for supervision and corresponding research.
Key words: Prohibited drug; HPLC /MS; Chinese herb extract
In recent years, there have been reports of illegal addition of synthetic drugs in health foods or plant extracts, including dozens of drugs such as various stimulants, diuretics, aphrodisiacs, anorexia agents, etc., in order to protect the health of consumers. The country has established relevant technical standards. In this analysis process, three aspects need to be solved. First, the scope of analysis of drugs is very large, especially in terms of polarity, acidity and alkalinity, how to achieve one analysis and complete spectrum work; second, drugs The content limits vary greatly, some allow only a few milligrams per kilogram; the third is how to accurately complete qualitative conclusions. Based on foreign relevant experimental standards, the Waters Quattro LC system combined with the Waters Symmetry C18 column has been used to establish a complete corresponding illicit drug analysis system.
1 Instrument conditions 1. 1 Chromatographic conditions Chromatographic conditions: Waters 2690 liquid chromatography system / Waters 996PAD detector, the column is Waters Symmetry C18 (2.1mm × 150mm), the mobile phase is 0.1% HAc / H2O (A), pure CH3CN (B), using gradient elution: 0 min, 3% B; 0-6 min, 3% B; 6-52 min, linear to 95% B; 52-60 min, 95% B; 60-61 min, linear to 3% B, the running time was 73 min, the flow rate was 0.25 mL / min, and the injection volume was 20 μL.
2 Mass Spectrometry Conditions Micromass Quatrro LC Mass Spectrometer (LC/MS/MS), Source: ES(+), Corona: 3.50kvolts, Cone: 20volts, Extractor: 3volts, Source block temp: 100°C, Desolvation temp: 450°C, Collision: 15evolts, Neb gas flow: 60L / h, Desol gas flow: 430L / h, Multiplier: 650volts.
2 Sample treatment Take 1.0g of solid sample, add 1.0ml of methanol, extract it in ultrasonic for 20min, ultracentrifugation for 10min, take the supernatant for LC/MS analysis, and directly filter the liquid sample by centrifugation and 0.22μm filter membrane. Perform LC/MS analysis.
3 Results and discussion 3. 1 Chromatographic retention Due to the large differences in the chemical nature of the compounds to be analyzed, from polar to non-polar, from acidic to basic, the relevant liquid-quality analysis is carried out, and good separation on the column is The basis of this analysis. We used Water Symmetry C18 (2.1mm × 150mm) and organic solvent gradient elution in acid suppression mode to achieve complete baseline separation of all target peaks, even with a retention time of only 0.2min. The basic separation indicates that the Symmetry column has excellent analytical capabilities.
3. 2 Recognition of retention time by chromatographic mass spectrometry While chromatographic separation and spectral identification are carried out, the mass chromatogram of liquid chromatography/mass spectrometer is preferred to further identify and confirm the retention time of the corresponding substance. And obtain the mass spectrum of the corresponding peak position of each component, refer to the standard retention time (RT) and molecular ion peak information as one of the primary conditions for qualitative determination of each component in the sample.
3 Confirmation of drug components 3. 1 LC/MS
After the sample is analyzed by LC/MS, the mass chromatogram of the sample is extracted according to the molecular ion peak of the reference or the molecular ion peak derived theoretically, and the compound of the reference product can have the standard RT as the reference. For the preliminary determination of the positive compound; for the compound without the control, because of the interference of the sample matrix, there are often several peaks at the same mass-to-charge ratio. In this case, only the corresponding mass spectrometry information and some characteristics of the binding compound itself are used. Judging, especially in combination with characteristic isotope peaks, peak intensity of ions (the intensity of the general mass chromatogram is greater than 10 to the 5th power) and other factors are considered.
3. 2 LC/MS/MS
After preliminary analysis by LC/MS, for the chromatographic peaks that are initially judged to be positive, MS/MS low energy collision method (15evolts) is used to scan the corresponding specific ion ions to obtain the “daughter†fragment mass spectrum and The zui final confirmation is performed by comparison with the fragment mass spectrum of the control or by comparison with the theoretical structure derivation. For example, in this analysis, the peak-to-mass ratio of a peak at a retention time of 18.2 min is 279, but no reference substance can be compared. It is suspected to be butylamine furosemide by other means. We found through the characteristic fragmentation of the peak. Its lysis method is confirmed by butylamine furosemide.
We use Waters Symmetry C18, Waters 2690 LC system and Quattro LC LC/MS to establish a complete and corresponding analysis system for dozens of illicit drugs. It has fast and reliable characteristics and can meet daily inspection and regulations. Inspection requirements.
Increases fermentable carbohydrates
Produces highly attenuated beer in a cost-effective manner
Higher extract yield
Abstract: [Objective] To establish a screening method for the illegal addition of synthetic drugs in proprietary Chinese medicines. [Method] Various kinds of added drugs were extracted by organic solvent, and the liquid chromatography- mass spectrometer was used for measurement. For some samples with positive primary mass spectrometry results, a multi-stage mass spectrometric structural study was carried out to confirm the compounds. [Results] Based on the retention time, molecular ion mass and multi-stage mass spectrometry results of various drugs, a screening method for the determination of 65 drugs was established. [Conclusion] This method established a mass spectrometry screening and analysis system for various drugs, which provided technical guarantee for supervision and related research.