The short-day condition significantly affected the formation of bolting and the thickening and morphology of rhizome of Alisma orientale. Therefore, it was necessary to conduct the sowing and raising of seedlings in the high-temperature season. The seeds were sown on cell trays containing field soil on Jul. 17 (plot S-7.17), Aug. 12 (plot S-8.12) and Sep. 11 (plot S-9.11) in 2013, germinated in a few days and developed to juvenile plants to be settled for 2 months. These juvenile plants cultivated for 4 months from settled planting, formed 5.0±1.2 scapes at plot S-7.17 and 0.6±0.8 scapes at plot S-8.12; however, no scapes were formed at plot S-9.11. The rhizome at plot S-7.17 was the highest in dry weight (63.2±7.6g), and composed of a large amorphous rhizome and several small rhizomes, whereas, the plant at plot S-9.11 formed a large oval rhizome. All dry rhizomes at each experimental plot contained alisol A, alisol B and alisol B monoacetate. In future, based on these results the cultivation of A. orientale will be conducted on paddy fields in a warm region in Japan.

On Iriomote Island, Okinawa Pref., Japan, we conducted cultivation experiments for Alisma orientale. The seeds were sown on cell trays containing field soil on Jul. 17 (plot I-7.17), Aug. 12 in 2013 (plot I-8.12) and Sep. 16, 2014 (plot I-2015), and developed to juvenile plants to be settled. On Oct. 4 (plot I-7.17) and Oct. 30 in 2013 (plot I-8.12), we settled juvenile plants in 1/2000a Wagner pots containing field soil, and then cultivated them for 3 months until harvest. The mature plants bearing scapes numbered 5 out of 10 individuals at plot I-7.17; however, no scapes were formed at plot I-8.12. On Dec. 24, 2014 (plot I-2015), 300 juvenile plants were planted in the paddy field, and cultivated for 4 months until harvest. None of the mature plants at plot I-2015 formed scape, and a large oval rhizome was harvested. The processed rhizomes after harvesting were similar to those of Alisma Tuber (沢瀉) imported from China in morphology, and contained alisol A, alisol B and alisol B monoacetate.

A TLC-based method was developed for the identification of the crude drug Angelicae Dahuricae Radix (Angelica Dahurica Root; root of Angelica dahurica Bentham et Hooker filius ex Franchet et Savatier). UV-sensitive constituents in 11 samples of Angelicae Dahuricae Radix available on the market in Japan were compared using TLC. Six common spots were detected in all samples and these were isolated and identified as isoimperatorin，phellopterin，imperatorin，byakangelicol，oxypeucedanin hydrate, and byakangelicin based on the results of spectroscopic methods. Among them, two compounds, oxypeucedanin hydrate and byakangelicin, were concluded to be characteristic of Angelicae Dahuricae Radix. The present data could be useful for the development of convenient TLC tests for identifying Angelicae Dahuricae Radix.

The origin of Arecae Semen is a seed of Areca catechu Linné (Palmae) and it is used as an ingredient of Kampo formulae such as Kumibinroto and Nyoshinsan. The seed contains alkaloids such as arecaidine, arecoline, guvacoline and guvacine. Arecaidine, arecoline and guvacine in Arecae Semen can be analyzed on HPLC with solvent containing sodium lauryl sulfate as a mobile phase. However, this method is not very convenient mainly because of the bubbles of sodium lauryl sulfate. In this study, a new method for HPLC analyses of arecaidine, arecoline and guvacine in Arecae Semen without the use of sodium lauryl sulfate was investigated. Finally, a new method using perchloric acid as a mobile phase was developed.

Cinnamon Bark (“Keihi”) is an important crude drug used in Kampo-medicine. Recently, powdered cinnamon bark formulation was approved as an “efficacious treatment for dry mouth, thirst, and diabetes,” and it became possible to commercialize the product as an over-the-counter (OTC) agent. To obtain this approval, the examination of quality control-related characteristics is necessary and, so, we considered developing a testing method. Therefore, we established a quantitative method for (E)-cinnamic acid and coumarin determination. In particular, it is important to establish a quantitative method for coumarin because its association with hepatic injury is a concern.

The origin of Bezoar Bovis (BB) is a stone formed in the gall sac of Bos taurus Linné var. domesticus Gmelin (Bovidae), which is designated in the Japanese Pharmacopoeia Seventeenth Edition (JP17). It is important to determine the quantity of its principal component, bilirubin, for the BB quality evaluation. Although a semi-quantitative method for measuring chloroform-soluble BB ingredients including bilirubin is available, a quantitative assay for bilirubin is not described in JP17. Furthermore, from the viewpoint of clean analysis, it is undesirable that chloroform is used as the solvent for the identification test of BB in JP17. In our previous paper in 2010, we reported a HPLC-based quantification of bilirubin in BB that excluded chloroform and used a column packed with octylsilanized silica gel (C₈). In the present study, we improved the analytical conditions and established the quantitative HPLC method of bilirubin in BB with widely-used octadesylsilanized silica gel (C₁₈) to achieve constancy of extraction efficiency and stability of standard solution.