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Example 31
Step 1: 3-(Ethylamino)-1-methylpyrrolidin-2-one
A solution of 3-amino-1-methylpyrrolidin-2-one (360 mg, 3.2 mmol) and acetaldehyde (0.89 mL, 15.8 mmol) was stirred at r.t. for 2 h in DCM (7.5 mL). Upon full conversion of amine as detected by LCMS (pH 10), the reaction mixture was concentrated under reduced pressure. Then the residue was redissolved in DCM (7.5 mL) and methanol (2.5 mL), at which point sodium cyanoborohydride (400 mg, 6.3 mmol) was added and the reaction mixture was stirred at room temperature for 16 h. The reaction was diluted with saturated NaHCO3 and the organics were extracted with EtOAc (3×). The combined organics were dried over MgSO4 and concentrated. The crude product was taken forward without additional purification. LCMS calculated for C7H15N2O (M+H)+: 143.1; found 143.2.
Example 22
Clinicians can use several biochemical measurements to objectively assess patients' current or past alcohol use. Several more experimental markers hold promise for measuring acute alcohol consumption and relapse. These include certain alcohol byproducts, such as acetaldehyde, ethyl glucuronide (EtG), and fatty acid ethyl esters (FAEE), as well as two measures of sialic acid, a carbohydrate that appears to be altered in alcoholics (Peterson K, Alcohol Research and Health, 2005). Clinicians have had access to a group of biomarkers that indicate a person's alcohol intake. Several of these reflect the activity of certain liver enzymes: serum gamma-glutamyltransferase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and carbohydrate-deficient transferrin (CDT), a protein that has received much attention in recent years. Another marker, N-acetyl-β-hexosaminidase (beta-Hex), indicates that liver cells, as well as other cells, have been breaking down carbohydrates, which are found in great numbers in alcohol (Javors and Johnson 2003).
In some embodiments the disclosed device focuses on detecting markers associated with alcohol abuse from menstrual blood or cervicovaginal fluid.