The Impact of Green Tea Extract (GTE) on the Metabolism of Clomipramine in Rat Liver Microsome Using HPLC and Compact Mass Spectrometry
Abstract
Clomipramine is a commonly used tricyclic antidepressant in treating a variety of disorders such as obsessive-compulsive disorder, anxiety, depression, and panic disorders. Clomipramine is metabolized primarily by the isoenzyme CYP450 2D6 in the liver. It can also be metabolized by three other enzymes, CYP450 1A2, CYP450 2C19, and CYP450 3A4 isoenzymes. These are four isoenzymes for metabolizing Clomipramine, but the focus for this experiment is CYP450 1A2 with rat liver microsome using HPLC and compact mass spectrometry (CMS). Today, roughly $11 billion is spent on green tea consumption. One form of consuming green tea is through green tea extract. Green tea extract, in a pill or powder form, contains high amounts of antioxidants, polyphenols, minerals, caffeine, carbohydrates, and various amounts of vitamins. The question of interest is "how much is too much?" for clomipramine users. This experiment explores green tea extract in a powder and the effects it has on the metabolization of clomipramine. Green tea extract is known to inhibit two metabolizing enzymes, CYP450 1A2 and CYP450 2D6, but it is unknown which is primarily inhibited. To carry out the experiment, four negative controls and various concentrations of green tea extracts were used. The negative controls contained varying amounts of clomipramine, NADPH, microsomal suspension, and buffer. The experimental samples contained varying amounts of green tea extract (inhibitor) in addition to clomipramine, microsome, and NADPH. The samples were incubated at 37 °C . The metabolite was extracted and mixed with the mobile phase to be injected through a compact mass spectrometer fitted with a C18 reverse phase column. The CMS data showed a 51.4% decreased level of desmethylated clomipramine, indicating that the CYP450 1A2 isoenzyme is being inhibited. From the data gathered through CMS, a graphical analysis determined the concentration of green tea extract at a 50% inhibition under the experimental condition. These results showed that IC50 was 28.3 mg/mL green tea extract under the experimental condition. This can be significant for informing individuals who consume green tea extract and clomipramine together.
College
College of Science & Engineering
Department
Chemistry
Campus
Winona
First Advisor/Mentor
Myoung Lee
Start Date
4-19-2023 2:00 PM
End Date
4-19-2023 3:00 PM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Session
2b=2pm-3pm
Poster Number
30
The Impact of Green Tea Extract (GTE) on the Metabolism of Clomipramine in Rat Liver Microsome Using HPLC and Compact Mass Spectrometry
Clomipramine is a commonly used tricyclic antidepressant in treating a variety of disorders such as obsessive-compulsive disorder, anxiety, depression, and panic disorders. Clomipramine is metabolized primarily by the isoenzyme CYP450 2D6 in the liver. It can also be metabolized by three other enzymes, CYP450 1A2, CYP450 2C19, and CYP450 3A4 isoenzymes. These are four isoenzymes for metabolizing Clomipramine, but the focus for this experiment is CYP450 1A2 with rat liver microsome using HPLC and compact mass spectrometry (CMS). Today, roughly $11 billion is spent on green tea consumption. One form of consuming green tea is through green tea extract. Green tea extract, in a pill or powder form, contains high amounts of antioxidants, polyphenols, minerals, caffeine, carbohydrates, and various amounts of vitamins. The question of interest is "how much is too much?" for clomipramine users. This experiment explores green tea extract in a powder and the effects it has on the metabolization of clomipramine. Green tea extract is known to inhibit two metabolizing enzymes, CYP450 1A2 and CYP450 2D6, but it is unknown which is primarily inhibited. To carry out the experiment, four negative controls and various concentrations of green tea extracts were used. The negative controls contained varying amounts of clomipramine, NADPH, microsomal suspension, and buffer. The experimental samples contained varying amounts of green tea extract (inhibitor) in addition to clomipramine, microsome, and NADPH. The samples were incubated at 37 °C . The metabolite was extracted and mixed with the mobile phase to be injected through a compact mass spectrometer fitted with a C18 reverse phase column. The CMS data showed a 51.4% decreased level of desmethylated clomipramine, indicating that the CYP450 1A2 isoenzyme is being inhibited. From the data gathered through CMS, a graphical analysis determined the concentration of green tea extract at a 50% inhibition under the experimental condition. These results showed that IC50 was 28.3 mg/mL green tea extract under the experimental condition. This can be significant for informing individuals who consume green tea extract and clomipramine together.
