Identification and estimation of the levo isomer in raw materials and finished products containing atropine and/or hyoscyamine.
The belladonna alkaloids atropine sulfate and hyoscyamine sulfate, occasionally used as anticholinergic and antimuscarinic agents, have identical molecular formulas but different stereo configurations. Hyoscyamine sulfate contains almost 100% of the levo isomer, whereas atropine sulfate is composed of equal parts of dextro and levo isomers. It is believed that the therapeutic properties of these alkaloids are due exclusively or primarily to the levo isomer. Currently available methods determine only the total amount of atropine (hyoscyamine) sulfate. A method has been developed and is reported for the identification and estimation of the levo and dextro isomers of atropine and hyoscyamine. Reference solutions are prepared in methanol at the following weights per 100 mL: 8.0 mg atropine sulfate; 4.0 mg hyoscyamine sulfate; 7.0 mg scopolamine hydrobromide; and 10.0 mg homatropine methylbromide. Samples of raw materials are similarly prepared in methanol, commercial products are also extracted or diluted with methanol, and solutions are filtered. Liquid chromatography is used for separations on a 25 cm Chirobiotic T2 column. The mobile phase is prepared by mixing 3.0 mL acetic acid and 2.0 mL triethylamine with 1000 mL methanol. The injection volume is 100 or 200 microL; the flow rate is about 0.35 mL/min. Fluorescence detection is at 255 nm excitation and 285 nm emission. Scopolamine hydrobromide and hyoscyamine eluted after 20 and 60 min, respectively. Atropine sulfate generated 2 peaks after 60 and 65 min. Homatropine methylbromide also produced 2 peaks after 70 and 85 min. Samples tested in this study included raw materials and commercial tablets or injections containing belladonna alkaloids. In all cases, the percentage calculated was that of the levo isomer relative to the total amount of atropine (hyoscyamine) present.
Cieri
Ugo R
UR
U.S. Food and Drug Administration, 2nd and Chestnut Sts, Philadelphia, PA 19106, USA. ucieri@ora.fda.gov
D001285:Atropine / Q000032:analysis* / Q000737:chemistry*; D001533:Belladonna Alkaloids / Q000032:analysis / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002845:Chromatography; D002853:Chromatography, Liquid; D000432:Methanol / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism
D001533:Belladonna Alkaloids; D001285:Atropine; D000432:Methanol