Herbal Textual Research on Traditional Chinese Medicines of Doukou in Famous Classical Formulas / 中国实验方剂学杂志

Chinese medicines of Doukou includes Amomi Fructus Rotundus， Alpiniae Katsumadai Semen， Galangae Fructus and Myristicae Semen. They have a long medicinal history and are also commonly used in cooking and seasoning. Due to the similar names and limited to the traffic conditions in ancient times， the records of Doukou in ancient literature are often confused with many plants in the same family， and there are still many kinds of confused products. In order to promote the development of famous classical formulas containing the medicinal materials， the ancient literature of Doukou in the past dynasties was comprehensively combed from the aspects of name， origin， genuine area， medicinal parts， harvesting and processing and processing methods. It has been found that the basic original plants of Amomi Fructus Rotundus are Amomum kravanh and A. compactum， the original plant of Alpiniae Katsumadai Semen is Alpinia katsumadai and it often confused with Tsaoko Fructus. The main source of Galangae Fructus recorded in the ancient materia medica is the fruit of A. officinarum， while the 2020 edition of Chinese Pharmacopoeia stipulates that the original plant is A. galanga. Myristica fragrans is the original plant of Myristicae Semen. It was found that except M. fragrans， the other three kinds of medicinal origin of Doukou had changed， there are many other plants confused with each other. The four kinds of Doukou are produced in Southeast China and Southeast Asia， and most of Amomi Fructus Rotundus and Myristicae Semen are imported. The Chinese medicines of Doukou have clear medicinal parts and simple processing methods， the main methods in the past dynasties are cleaning， stir frying and simmering， and the processed products are selected according to the needs of different diseases. It is suggested to use the dry mature seeds of A. katsumadai in Houpo Wenzhongtang， which is from Guangxi， Guangdong， Hainan， Fujian and Yunnan and so on， among which Wanning in Hainan province is genuine producing area. The fruits should be harvested in summer and autumn， and dried to 90% dry in the sun， or slightly scalded with water and dried to half dry in the sun， and removed the peel， taken out the seed group， dried in the sun and then be used as medicine.

High-speed identification of odor changes and substance basis of Myristicae Semen mildew by electronic nose and HS-GC-MS / 中国中药杂志

This study adopted headspace-gas chromatography-mass spectrometry(HS-GC-MS) and electronic nose to detect volatile components from Myristicae Semen samples with varying degrees of mildew, aiming at rapidly identifying odor changes and substance basis of Myristicae Semen mildew. The experimental data were analyzed by electronic nose and principal component analysis(PCA). The results showed that Myristicae Semen samples were divided into the following three categories by electronic nose and PCA: mildew-free samples, slightly mildewy samples, and mildewy samples. Myristicae Semen samples with different degrees of mildew greatly varied in volatile components. The volatile components in the samples were qualitatively and quantitatively detected by HS-GC-MS, and 59 compounds were obtained. There were significant differences in the composition and content in Myristicae Semen samples with different degrees of mildew. The PCA results were the same as those by electronic nose. Among them, 3-crene, D-limonene, and other terpenes were important indicators for the identification of mildew. Bicyclo[3.1.0]hexane, 4-methylene-1-(1-methylethyl)-, terpinen-4-ol, and other alcohols were key substances to distinguish the degree of mildew. In the later stage of mildew, Myristicae Semen produced a small amount of hydroxyl and aldehyde compounds such as acetaldehyde, 2-methyl-propionaldehyde, 2-methyl-butyraldehyde, and formic acid, which were deduced as the material basis of the mildew. The results are expected to provide a basis for the rapid identification of Myristicae Semen with different degrees of mildew, odor changes, and the substance basis of mildew.

High-throughput screening of ochratoxin A in Chinese herbal medicines using enzyme-linked immunoassay / 中国中药杂志

An indirect competitive enzyme-linked immunosorbent assay( ic-ELISA) was developed for the rapid detection of ochratoxin A( OTA) in nutmeg( Myristicae Semen),ginger( Zingiberis Rhizoma) and turmeric( Curcumae Longae Rhizoma). The matrix matching standard curve was used instead of the standard curve of sample diluent,and the sample extract and sample diluent were optimized. The sensitivity( IC_(50)) of this method for OTA in nutmeg,ginger and turmeric were determined as 0. 146,0. 157 and 0. 153 ng·m L~(-1),respectively and the limits of detection( LODs) were 0. 040,0. 032 and 0. 031 ng·m L~(-1),respectively. The recovery of samples ranged from 75. 99% to 122. 3%,with RSD<10%. Two positive samples for nutmeg and one positive sample for turmeric occurred in 50 samples,and the highest OTA contamination value was 1 167. 8 μg·kg~(-1). The results were further confirmed by LC-MS/MS. It shows that the developed ic-ELISA method is simple,rapid and sensitive,and can be applied for rapid and high-throughput screening of OTA in nutmeg,ginger and turmeric,as well as some other CHMs.

Study on Purification and Anti-inflammatory Effects of Total Lignans in Myristicae Semen / 中国中医药信息杂志

Objective To optimize purification technology of total lignans in Myristicae Semen; To study its anti-inflammatory effects. Methods Macroporous adsorption resin was used for enrichment and purification of total lignans in Myristicae Semen, with adsorption rate, desorption rate and content of total lignans as indexes. The resin type, concentration of eluent, sample weight and dosage of eluen were investigated. Anti-inflammatory effects were studied by determining swelling degree and inhibition rate of mice with ear swelling induced by xylene. Results Purification technology of total lignans was processed on type of AB-8 resin. 1 g crude extract was dissolved as sample; 40 mL preprocessed resin including 16 g resin was added and static adsorpted for 12 hours; 30% ethanol was used firstly to elute to colorless and then 300 mL 70% ethanol was used to elute; the eluent of 70% ethanol was collected. The purity of total lignans was 45.8%. There was statistical significance in ear swelling degree of total lignans in Myristicae Semen high-, medium-, and low-dosage groups compared with model group (P<0.05). Conclusion The purification technology of total lignans is stable and reproducible, and total lignans in Myristicae Semen has significant anti-inflammatory activity.

Optimization of Bran-roasted Processing Technology of Sliced Myristicae Semen by Orthogonal Test / 中国中医药信息杂志

Objective To optimize the processing technology of sliced Myristicae Semen. Methods Roasting temperature, roasting time and the amount of bran were set as factors, and the content of total lignans, volatile oil, fatty oil were set as evaluation indicators. The processing technology of sliced Myristicae Semen was optimized by L9(34) orthogonal test. Results The optimal processing technology was as following: 40 g bran plus 100 g sliced Myristicae Semen, roasting for 20 minutes at 110-120 ℃. Conclusion The process is reasonable and reliable, which can provide references for new processing technology of Myristicae Semen.

Optimization of ethanol extraction process of Psoraleae Fructus-Myristicae Semen drug pair by orthogonal test combined with fingerprint / 中草药

Objective: To optimize the ethanol extraction process of Psoraleae Fructus-Myristicae Semen (psoralen-nutmeg) drug pair. Methods: Using L9(34) orthogonal design, the effects of ethanol concentration, ethanol amount, extraction time, and extraction times on the extraction process were investigated. The contents of psoralen, isopsoralen, and dehydrodiisoeugenol, dry extract yield, and total area of HPLC fingerprint characteristic peaks were used as comprehensive evaluation indexes. Results: The optimum process conditions were as follows: 50% ethanol, six times of the ethanol volume, extracted for three times, each time for 2 h. Conclusion: The method provides the basis for the determination of ethanol extraction process of Psoraleae Fructus-Myristicae Semen drug pair.

Study on chemical constituents of Myristicae Semen / 中草药

Objective: To study the chemical constituents in the dry mature seeds of Myristica fragrans (Myristicae Semen). Methods: Compounds were isolated and purified from the ethyl acetate layer of 75% alcohol extract by means of the silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC isolation methods. Compound structures were identified by analyzing and comparing the spectral data with those for them in the reference. Results: Eighteen compounds were obtained from the ethyl acetate layer of 75% ethanol extract in Myristicae Semen. On the basis of spectral data and combined with the references, these compounds had been identifited as: vanilic acid (1), butein (2), (2R)-3-(3', 4', 5'-trimethoxyphenyl)-1, 2- propanediol (3), sulphuretin (4), 3-methoxy-4, 5-methylenedioxy-cinnamic acid (5), 7, 3', 4'-trihydroxyflavanone (6), 7-hydroxy- 4-benzopyrone (7), verrucosin (8), (+)-erythro-(7S, 8R)-Δ8'-7-hydroxy-3, 4, 3', 5'-tetramethoxy-8-O-4'-neolignan (9), (-)-erythro-(7R, 8S)-Δ8'-7- acetoxy-3, 4, 3', 5'-tetramethoxy-8-O-4'-neolignan (10), nectandrin B (11), (-)-(7S, 7'R, 8S, 8'R)-4, 4'-dihydroxy-3, 5, 3'-trimethoxy-7, 7'-epoxylignan (12), fragransin B3 (13), fragransin B1 (14), (-)-enantiomer (15), (-)-erythro-(7R, 8S)-Δ8'-7-hydroxy-3, 4, 5, 3', 5'-pentamethoxy-8-O-4'-neolignan (16), (+)-erythro-(7S, 8R)-Δ8'-7, 4-dihydroxy-3, 5, 3', 5'-tetramethoxy-8-O-4'-neolignan (17), and (+)-5-methoxydeydrodiisoeugenol (18). Conclusion: Compounds 2 and 4-7 are isolated from the plants of Myristica Gronov. for the first time. Compound 1 is isolated from this plant for the first time.