Hoshi University

A systematic analytical method for discriminating methamphetamine (MA) from its structural analogs was developed by combining a solid-phase chromogenic method with the use of a molecularly imprinted polymer (MIP) and Simon reaction. L-proline (Pro), L-hydroxyproline (HYP), N-methylbenzylamine (NMe-BA), and N-isopropylbenzylamine (NIP-BA) were selected as false positives, while tert-butoxycarbonyl derivatized MA (t-Boc-MA) was selected as a false-negative substance. MA and the structural analogs were loaded into SupelMIP™ SPE-Amphetamines cartridges and Simon's reagent was added to the cartridge, then the coloration of the solid-phase gel was observed from outside the cartridge. After elution with a basic solution, Simon's reagent was added to the eluate (first-step eluate), and a change in color tone was observed. The MIP was then heated under acidic conditions, and the analytes were eluted (second-step eluate) with water, basified at pH 10, and the Simon reaction was repeated. Pro and HYP could be discriminated from MA by the color tone of the MIP solid-phase gel, whereas NMe-BA and NIP-BA could be discriminated from MA by the color tone of the first-step eluate. In addition, the color tone of the second-step eluate confirmed the presence of t-Boc-MA.

Chronic constipation decreases quality of life; therefore, appropriate treatment measures are crucial. However, current treatments are insufficient to adequately control defecation, necessitating the identification of new strategies that are more effective and safer. We investigated the laxative action mechanism of the Kampo medicine Mashiningan (MNG) and discussed its usefulness in constipation. Rats were orally administered extracts of MNG or its herbal constituents, and defecation was analyzed. Expression levels of aquaporin-3 (AQP3) and inflammation-related genes in the colon were detected. The effects of the herbal medicines on inflammation were analyzed using macrophage-like RAW264.7 cells. Fecal water content significantly increased in rats treated with MNG. Expression of AQP3 protein in the colon of the MNG-treated group reduced to ∼20 % of that in the control group. Examining the effect of compound herbal medicines, only administration of Daio extract increased defecation and reduced colonic expression of AQP3. Treatment with MNG significantly increased the expression of interleukin (IL)-1β and IL-6 in the colon, although this increase was less than that in the group treated with Daio alone. Mashinin, Kyonin, and Kijitsu could significantly suppress the lipopolysaccharide (LPS)-induced increase in IL-1β and IL-6 in RAW264.7 cells. MNG exerted a laxative effect by decreasing AQP3 expression and disturbing water absorption in the colon. This effect was attributed to the herbal compound Daio. Furthermore, MNG contains Mashinin, Kyonin, and Kijitsu, which exert anti-inflammatory effects. Accordingly, MNG is a laxative with fewer side effects, such as abdominal pain.

Oversulfated chondroitin sulfate (OSCS) is a chondroitin sulfate ester in which all hydroxyl groups have been converted to sulfuric acid esters. Here, we describe a rapid, novel synthesis method for OSCS. ¹H-NMR analysis revealed that all signals derived from the protons of N-acetyl-galactosamine and glucuronic acid in the synthesized OSCS were shifted downfield due to sulfonation compared with the chondroitin sulfate (CS) starting material. Comparison of the edited heteronuclear single quantum correlation spectrum of the prepared OSCS with that of the Japanese Pharmacopoeia OSCS reference standard showed excellent agreement between the main correlation peaks. The proposed novel synthesis method for OSCS is faster and simpler than the conventional method.