Study on triterpenoic acids distribution in Ganoderma mushrooms by automatic multiple development high performance thin layer chromatographic fingerprint analysis.

Ganoderma--"Lingzhi" in Chinese--is one of the superior Chinese tonic materia medicas in China, Japan, and Korea. Two species, Ganoderma lucidum (Red Lingzhi) and G. sinense (Purple Lingzhi), have been included in the Chinese Pharmacopoeia since its 2000 Edition. However, some other species of Ganoderma are also available in the market. For example, there are five species divided by color called "Penta-colors Lingzhi" that have been advocated as being the most invigorating among the Lingzhi species; but there is no scientific evidence for such a claim. Morphological identification can serve as an effective practice for differentiating the various species, but the inherent quality has to be delineated by chemical analysis. Among the diverse constituents in Lingzhi, triterpenoids are commonly recognized as the major active ingredients. An automatic triple development HPTLC fingerprint analysis was carried out for detecting the distribution consistency of the triterpenoic acids in various Lingzhi samples. The chromatographic conditions were optimized as follows: stationary phase, precoated HPTLC silica gel 60 plate; mobile phase, toluene-ethyl acetate-methanol-formic acid (15 + 15 + 1 + 0.1); and triple-development using automatic multiple development equipment. The chromatograms showed good resolution, and the color images provided more specific HPTLC fingerprints than have been previously published. It was observed that the abundance of triterpenoic acids and consistent fingerprint pattern in Red Lingzhi (fruiting body of G. lucidum) outweighs the other species of Lingzhi.

Chemical pattern-aided classification to simplify the intricacy of morphological taxonomy of Epimedium species using chromatographic fingerprinting.

Epimedium herb (Yinyanghuo), one of the popular Chinese materia medica, is a multiple species colony of Epimedium genus belonging to Berberidaceae. There are five species of Epimedium that have been officially adopted in Chinese Pharmacopoeia under the same crude drug name 'Yinyanghuo' comprising Epimedium brevicornu, E. koreanum, E. sagittatum, E. pubescens, and E. wushanense. In addition, non-official species like E. acuminatum, E. miryanthum and E. leptorrhizum are also mix-used. Frequently, the morphological taxonomical identification is very difficult during on-site inspection for species authentication in the market. Researchers are often bewildered by the multiple species ambiguity when putting this crude drug in use. Referring to the bioactive constituents that are vital for therapeutic efficacy, the key to clarifying the multiple species confusion should rely on analysis of the bioactive composition. It is well known that medicinal Epimedium herbs contain special C-8 prenylated flavonol glycosides which contribute to various bioactivities and the major four, epimedin A (A), epimedin B (B), epimedin C (C) and icariin (I), are unanimously used as bioactive markers for quality control. In this study, HPLC-DAD fingerprinting was performed for investigating the molecular spectrum of various Epimedium species. It was found that the four major flavonoids constitute the middle part of the chromatographic profiles to form a specific region (named as 'ABCI fingerprint region') being dominant in the HPLC profiles of all medicinal Epimedium species, and the five official species express five different 'ABCI' patterns (different peak: peak ratios). Our study found that the convergent tendency of the 'ABCI region' among multiple species of Epimedium could facilitate differentiation of complex commercial samples based on similar bioactive composition should confer similar bioactivities. Merging the different species that possess the same 'ABCI region' pattern into the same group can create a simpler bioactive-fraction-aided classification array by clustering the commercial samples into three bioactive ingredients-based fingerprint patterns - 'E.b. pattern', 'E.k. pattern' and 'extensive E.w. pattern'. This approach offers the feasibility of characterizing and quality-controlling complex samples in the same genus designated under a single herbal drug entity on the premise of possessing the same bioactive ingredients pattern and supported by long-term traditional usage.

Study of the destructive effect to inherent quality of Angelicae dahuricae radix (Baizhi) by sulfur-fumigated process using chromatographic fingerprinting analysis.

The after-harvesting sun-dried process of Angelicae dahuricae radix (Chinese name: Baizhi) was previously the traditional treatment for commodity. Over recent decades the natural drying process for some fleshy roots or rhizomes of Chinese materia medica has been replaced by sulfur-fumigation for curtailing the drying duration and pest control. We used high performance liquid chromatography (HPLC) and high performance thin-layer chromatography (HPTLC) fingerprinting analysis to investigate the potential damaging effect of the sulfur-fumigating process. The experimental conditions were as follows. HPTLC analysis was carried out on pre-coated silica-gel 60 plate, twice development was performed with two solvent systems (mobile phase) A, chloroform-ethyl acetate (10:1) and B, hexane-chloroform-ether (4:1:2); the fluorescent images were observed under UV 365 nm. HPLC was preceeded on Zorbax SB-C(18) column; the linear gradient elution was conducted with mobile phase prepared from methanol-0.5% acetic acid; column temperature was at 25 degrees C; the detection wavelength was 250 nm. We found serious degradation of the majority of coumarins in sulfur-fumigated Baizhi. The destructive effect was manifested by the defaced chromatographic profile and verified by imitating the sulfur dioxide reaction with the constituents in Baizhi in the laboratory. It is suggested that sulfur-fumigation process is an unacceptable approach for processing herbal drugs.

Effect of Xuesaitong soft capsule on hemorrheology and in auxiliarily treating patients with acute cerebral infarction.

OBJECTIVE: To observe the therapeutic effect of Xuesaitong soft capsule (XST) and its effect on platelet counts, coagulation factor 1 (CF1) as well as hemorrheologic indexes in treating patients with acute cerebral infarction (ACI). METHODS: Two hundred and four patients with ACI were assigned into two groups, the control group (n = 96) and the treated group (n = 108). They were all treated with conventional Western medicines, including mannitol, troxerutin, citicoline, piracetam and aspirin, while to the treated group, XST was given additionally through oral intake, twice a day, 2 capsules each time for 8 successive weeks. The clinical efficacy was evaluated according to the nerve function deficits scoring and the changes of platelet count. CF1 and hemorrheological indexes were measured before and after treatment. RESULTS: The total effective rate was 87.0% in the treated group, and 87.5% in the control group, respectively, showing insignificant difference between them. But the markedly effective rate in the treated group (66.7%) was significantly higher than that in the control group (27.1%, P < 0.01). The count of platelet was not changed significantly in both groups after treatment, while CF1 in them evidently lowered at the end of the 4th and 8th weeks of treatment, but showed insignificant difference between the two groups. The hematocrit, whole blood viscosity and plasma viscosity in both groups were all improved significantly after treatment, but also showed insignificant difference in comparison of the two groups. CONCLUSION: XST has good efficacy in auxiliary treatment of patients with ACI, though its mechanism remains to be further explored.