Macrozamin

[Rapid determination of the multi-marker ingredients in Heterosmilacis Japonicae Rhizoma and Sophorae Flavescentis Radix with near-infrared diffused reflection spectroscopy].[Pubmed:25739202]

Guang Pu Xue Yu Guang Pu Fen Xi. 2014 Oct;34(10):2652-6.

A rapid NIRS method for determination of Macrozamin in Heterosmilacis japonicae rhizoma (HJR), and the total content of oxymatrine and matrine (OMT + MT) as well as the total content of oxysophocarpine and sophocarpine (OSC + SC) in sophorae flavescens radix (SFR) was developed to explore the application feasibility of NIRS for the quality assurance system of Chinese patent drugs. The contents of Macrozamin in HJR samples, and OMT + MT and OSC + SC in SFR samples were determined by HPLC as reference values. The NIR spectra of the samples were measured in a diffused reflection mode. The different characteristic wavebands and pretreatment methods were optimized. The quantitative calibration models between the NIR spectra and the content reference values of marker components in HJR and SFR samples, were established with partial least square method, and further optimized through the cross validation and external validation. The contents of Macrozamin in 88 batches of HJR samples were over the range of 0.36-12.88 mg . g(-1). The total contents of OMT + MT and OSC + SC in 75 batches of SFR samples were over the range of 8.87-66.31 and 2.30-15.11 mg . g(-1), respectively. The performance of the final models for Macrozamin, OMT + MT and OSC + SC was evaluated well according to correlation coefficients (r), root mean square error of cross-validation (RMSECV) and root mean square error of prediction (RMSEP). The R2 values of the cross-validation for Macrozamin, OMT + MT and OSC + SC were 0.9025, 0.9491 and 0.9137, and those of RMSECV were 0.961, 2.45 and 0.724 mg . g(-1) respectively. The R2 values of external validation for the three models were 0.9817, 0.9826 and 0.9609, and those of RMSEP were 0.693, 2.27 and 0.658 mg . g(-1), respectively. This is the first report on rapid determination of Macrozamin in Heterosmilacis japonicae rhizoma, and oxymatrine, matrine, oxysophocarpine and sophocarpine in sophorae flavescens radix by NIRS method. The presented method can fulfill the requirement of rapid acquirement of chemical information of raw medicinal materials prior the manufacturing of compound Kushen injection.

Animal models of brain maldevelopment induced by cycad plant genotoxins.[Pubmed:24339036]

Birth Defects Res C Embryo Today. 2013 Dec;99(4):247-55.

Cycads are long-lived tropical and subtropical plants that contain azoxyglycosides (e.g., cycasin, Macrozamin) and neurotoxic amino acids (notably beta-N-methylamino-l-alanine l-BMAA), toxins that have been implicated in the etiology of a disappearing neurodegenerative disease, amyotrophic lateral sclerosis and parkinsonism-dementia complex that has been present in high incidence among three genetically distinct populations in the western Pacific. The neuropathology of amyotrophic lateral sclerosis/parkinsonism-dementia complex includes features suggestive of brain maldevelopment, an experimentally proven property of cycasin attributable to the genotoxic action of its aglycone methylazoxymethanol (MAM). This property of MAM has been exploited by neurobiologists as a tool to study perturbations of brain development. Depending on the neurodevelopmental stage, MAM can induce features in laboratory animals that model certain characteristics of epilepsy, schizophrenia, or ataxia. Studies in DNA repair-deficient mice show that MAM perturbs brain development through a DNA damage-mediated mechanism. The brain DNA lesions produced by systemic MAM appear to modulate the expression of genes that regulate neurodevelopment and contribute to neurodegeneration. Epigenetic changes (histone lysine methylation) have also been detected in the underdeveloped brain after MAM administration. The DNA damage and epigenetic changes produced by MAM and, perhaps by chemically related substances (e.g., nitrosamines, nitrosoureas, hydrazines), might be an important mechanism by which early-life exposure to genotoxicants can induce long-term brain dysfunction.

[Alcohol-purification technology and its particle sedimentation process in manufactory of Fufang Kushen injection].[Pubmed:22375388]

Zhongguo Zhong Yao Za Zhi. 2011 Nov;36(22):3108-13.

OBJECTIVE: Fufang Kushen injection was selected as the model drug, to optimize its alcohol-purification process and understand the characteristics of particle sedimentation process, and to investigate the feasibility of using process analytical technology (PAT) on traditional Chinese medicine (TCM) manufacturing. METHOD: Total alkaloids (calculated by matrine, oxymatrine, sophoridine and oxysophoridine) and Macrozamin were selected as quality evaluation markers to optimize the process of Fufang Kushen injection purification with alcohol. Process parameters of particulate formed in the alcohol-purification, such as the number, density and sedimentation velocity, were also determined to define the sedimentation time and well understand the process. RESULT: The purification process was optimized as that alcohol is added to the concentrated extract solution (drug material) to certain concentration for 2 times and deposited the alcohol-solution containing drug-material to sediment for some time, i.e. 60% alcohol deposited for 36 hours, filter and then 80% -90% alcohol deposited for 6 hours in turn. The content of total alkaloids was decreased a little during the depositing process. The average settling time of particles with the diameters of 10, 25 microm were 157.7, 25.2 h in the first alcohol-purified process, and 84.2, 13.5 h in the second alcohol-purified process, respectively. CONCLUSION: The optimized alcohol-purification process remains the marker compositions better and compared with the initial process, it's time saving and much economy. The manufacturing quality of TCM-injection can be controlled by process. PAT pattern must be designed under the well understanding of process of TCM production.

[Studies on key processes of Fufang Kushen injection].[Pubmed:21710724]

Zhongguo Zhong Yao Za Zhi. 2011 Mar;36(6):666-71.

OBJECTIVE: To study the key processes of Fufang Kushen injection for technical upgrading. METHOD: Total alkaloids (sum of matrine, oxymatrine, sophoridine and oxysophoridine) and Macrozamin were selected as quality evaluation markers. The key processes of percolation with acetic acid and discoloration with activated carbon were optimized by orthogonal experiment design, and process of purification with alcohol was investigated by single factor method. RESULT: The optimal condition of percolation process is as follows: the medicinal materials are soaked for 9 h with 4 times water containing 0.8% acetic acid, then percolation starts at flow-rate of 5 mL x min(-1) x kg(-1) and adding 2 times 0.8% acetic acid solution is added at the same velocity. Purification process is that the concentrated solution is precipitated by 60%, 80% and 90% alcohol in turn. Discoloration process is that 6 activated carbon is added into the solution which is heated at 60 degrees C for 20 minutes. CONCLUSION: The optimal extraction process is not only simple, saving the industrial cycle, reducing the potential risk, but also decreasing the acetic acid amount to guarantee the acid-insoluble ash as well as the functional ingredients.

Two genera of Aulacoscelinae beetles reflexively bleed azoxyglycosides found in their host cycads.[Pubmed:21656034]

J Chem Ecol. 2011 Jul;37(7):736-40.

Aulacoscelinae beetles have an ancient relationship with cycads (Cycadophyta: Zamiaceae), which contain highly toxic azoxyglycoside (AZG) compounds. How these "primitive" leaf beetles deal with such host-derived compounds remains largely unknown. Collections were made of adult Aulacoscelis appendiculata from Zamia cf. elegantissima in Panama, A. vogti from Dioon edule in Mexico, and Janbechynea paradoxa from Zamia boliviana in Bolivia. Total AZG levels were quantified in both cycad leaves and adult beetles by high performance liquid chromatography (HPLC). On average, cycad leaves contained between 0.5-0.8% AZG (frozen weight, FW), while adult beetles feeding on the same leaves contained even higher levels of the compounds (average 0.9-1.5% FW). High AZG levels were isolated from reflex bleeding secreted at the leg joints when beetles were disturbed. Nuclear magnetic resonance and mass spectroscopy identified two AZGs, cycasin and Macrozamin, in the reflex bleeding; this is the first account of potentially plant-derived compounds in secretions of the Aulacoscelinae. These data as well as the basal phylogenetic position of the Aulacoscelinae suggest that sequestration of plant secondary metabolites appeared early in leaf beetle evolution.

[Studies on chemical constituents from Rhizoma Heterosmilacis Japonicae].[Pubmed:18300492]

Zhong Yao Cai. 2007 Oct;30(10):1242-4.

Six compounds were isolated from Rhizoma Heterosmilacis Japonicae. They were identified as beta-sitosterol (I) dau-costerol (II),3,3',5,5'-tetrahydroxy-4'-methoxystilbene (III), naringenin (IV), alpha-L-butylsorbopyranoside (V) and Macrozamin (VI). The compounds III, IV, V and VI were isolated from this genus for the first time.

[Determination of macrozamin in Rhizoma Heterosmilacis Japonicae by RP-HPLC].[Pubmed:18051902]

Zhongguo Zhong Yao Za Zhi. 2007 Sep;32(18):1900-2.

OBJECTIVE: To develop a RP-HPLC method for the determination of the content of Macrozamin in Rhizoma Heterosmilacis Japonicae. METHOD: A Century C18 AQ column (4.6 mm x 250 mm, 5 microm) was used with the mobile phase consisted of water (4:96). The flow rate was 1.0 mL x min(-1). The detection wavelength was set at 215 nm, and the column temperature was 35 degrees C. RESULT: The calibration curve was linear (r = 0.999 8) in the range of 19.12 - 382.4 microg x mL(-1) for Macrozamin, the average recovery of the method was 99.5%, with RSD 2.1% (n = 9). CONCLUSION: This method can be used for the quality study of Rhizoma Heterosmilacis Japonicae.

Azoxyglycoside content and beta-glycosidase activities in leaves of various cycads.[Pubmed:15561189]

Phytochemistry. 2004 Dec;65(24):3243-7.

Azoxyglycoside contents in leaves of 32 cycad species belonging to 10 cycad genera and the seeds of 4 Encephalartos species were analyzed by HPLC with a YMC-PA03 amide column. Azoxyglycosides were detected in mature leaves of 14 cycad species including 2 Bowenia, 2 Lepidozamia, 1 Microcycas, and 1 Stangeria species, but not in mature leaves of 18 other cycad species; 2 of 3 Ceratozamia, 1 of 3 Cycas, 3 of 3 Dioon, 10 of 11 Encephalartos, 1 of 3 Macrozamia and 1 of 3 Zamia species analyzed. The ratios of beta-glycosidase activity toward cycasin and Macrozamin in extracts from the leaves of 9 species belonging to 9 genera were measured. The hydrolysis of cycasin was higher in the leaf extracts of Cycas revoluta, Bowenia spectabilis, Stangeria eriopus and Ceratozamia mexicana, whereas in Lepidozamia hopei, the hydrolysis levels of cycasin and Macrozamin were similar. On the other hand, activity toward Macrozamin was higher in Dioon edule, Encephalartos villosus, Macrozamia miquelii and Zamia fischeri. The hydrolytic activities in most species were estimated to be sufficient for the release of methylazoxymethanol in leaves analogous to the cyanogenesis of cyanogenic plants. Therefore, hydrolysis of azoxyglycosides by endogenous glycosidase in leaves seems to occur by accidental injury of leaves. However, in M. miquelii leaf extract, hydrolytic activity toward Macrozamin was high and the activity toward cycasin was very low, though only cycasin was found in the leaves of this species.

The magical and medicinal usage of Stangeria eriopus in South Africa.[Pubmed:7967657]

J Ethnopharmacol. 1994 Jul 8;43(2):67-72.

The underground caudex of the cycad Stangeria eriopus is used extensively by several ethnic groups in South Africa, mainly as an ingredient in magical potions but also as an emetic. An assessment of two main outlets showed that 3410 plants were sold in the month of July 1992; continued usage of this material now threatens the remaining plant populations. A proximate analysis of the caudex material gives high carbohydrate content with only small percentages of fat, protein, fibre and ash. An unusually high content of sodium sulphate may explain the efficacy of Stangeria-containing preparations as an emetic. The phytosterols sitosterol and stigmasterol are present in a 4:1 ratio while the fatty acid component comprises palmitic, oleic, stearic and arachidic acids. Twelve amino acids were identified in the material, including the non-protein amino acids beta-alanine, gamma-aminobutyric acid and pyroglutamic acid. The candidate neurotoxin beta-N-methylamino-L-alanine could not be detected but cycasin is present at the levels of 0.17% and 0.21% in fresh and dry caudex material, respectively and appears to be accompanied by the related toxin, Macrozamin.

Acquired chemical defense in the lycaenid butterfly,Eumaeus atala.[Pubmed:24271999]

J Chem Ecol. 1989 Apr;15(4):1133-46.

The lycaenid butterfly,Eumaeus atala, was found to contain cycasin, an azoxyglycoside, by thin-layer chromatography (TLC). Quantification of cycasin content in 10 individual freshly killed and frozen males and females, using capillary gas chromatography (GC), showed that cycasin contents of individual butterflies ranged from 0.21 to 0.51 mg (1.24-2.75% dry weight). A museum specimen ofE. atala of unknown age had undetectable amounts of cycasin by GC. GC showed that larval frass contained about 0.10% cycasin, which was not detectable by TLC. Cycasin in the host plant was not detectable by TLC but was detected by GC and found to be 0.02% dry weight. There was no Macrozamin, another azoxyglycoside characteristic of many cycads, in the butterfly or plant. Feeding trials with a colony of the ant,Camponotus abdominalis floridanus, showed that both cycasin and the adult ofE. atala were deterrent to the ants.

Notes on the toxicity and carcinogenicity of some South African cycad species with special reference to that of Encephalartos lanatus.[Pubmed:6355475]

J S Afr Vet Assoc. 1983 Mar;54(1):33-42.

The South African Encephalartos species which are known or have been shown experimentally to be toxic and/or carcinogenic are reviewed briefly. The kernel of E. lanatus, the toxic and carcinogenic principle of which is Macrozamin, is a potent hepatotoxin and carcinogen when fed to rats. Renal mesenchymal tumours were the most common neoplasm to be induced but hepatocellular carcinoma also developed in one animal. The potential danger to man and animals of ingestion of the seeds of Encephalartos species is emphasized.