Genomics-driven derivatization of the bioactive fungal sesterterpenoid variecolin: Creation of an unnatural analogue with improved anticancer properties

A biosynthetic gene cluster for the bioactive fungal sesterterpenoids variecolin ( 1) and variecolactone ( 2) was identified in Aspergillus aculeatus ATCC 16872. Heterologous production of 1 and 2 was achieved in Aspergillus oryzae by expressing the sesterterpene synthase VrcA and the cytochrome P450 VrcB. Intriguingly, the replacement of VrcB with homologous P450s from other fungal terpenoid pathways yielded three new variecolin analogues ( 5- 7). Analysis of the compounds' anticancer activity in vitro and in vivo revealed that although 5 and 1 had comparable activities, 5 was associated with significantly reduced toxic side effects in cancer-bearing mice, indicating its potentially broader therapeutic window. Our study describes the first tests of variecolin and its analogues in animals and demonstrates the utility of synthetic biology for creating molecules with improved biological activities.

Systematic identification of chemical forms of key terpene synthase in Cinnamomum camphora / 中国中药杂志

Cinnamomum camphora is an important economic tree species in China. According to the type and content of main components in the volatile oil of leaf, C. camphora were divided into five chemotypes, including borneol-type, camphor-type, linalool-type, cineole-type, and nerolidol-type. Terpene synthase(TPS) is the key enzyme for the formation of these compounds. Although several key enzyme genes have been identified, the biosynthetic pathway of(+)-borneol, which has the most economic value, has not been reported. In this study, nine terpenoid synthase genes CcTPS1-CcTPS9 were cloned through transcriptome analysis of four chemical-type leaves. After the recombinant protein was induced by Escherichia coli, geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) were used as substrates for enzymatic reaction, respectively. Both CcTPS1 and CcTPS9 could catalyze GPP to produce bornyl pyrophosphate, which could be hydrolyzed by phosphohydrolase to obtain(+)-borneol, and the product of(+)-borneol accounted for 0.4% and 89.3%, respectively. Both CcTPS3 and CcTPS6 could catalyze GPP to generate a single product linalool, and CcTPS6 could also react with FPP to generate nerolidol. CcTPS8 reacted with GPP to produce 1,8-cineol(30.71%). Nine terpene synthases produced 9 monoterpene and 6 sesquiterpenes. The study has identified the key enzyme genes responsible for borneol biosynthesis in C. camphora for the first time, laying a foundation for further elucidating the molecular mechanism of chemical type formation and cultivating new varieties of borneol with high yield by using bioengineering technology.

Identification of terpene synthase gene family in Gynostemma pentaphyllum and expression pattern analysis under abiotic stresses / 中国中药杂志

The present study aimed to investigate the composition of the terpene synthase(TPS) gene family in Gynostemma pentaphyllum and its role in abiotic stresses. The G. pentaphyllum TPS gene family was identified and analyzed at the genome-wide level using bioinformatics analysis, and the expression patterns of these family members were analyzed in different tissues of G. pentaphyllum as well as under various abiotic stresses. The results showed that there were 24 TPS gene family members in G. pentaphyllum with protein lengths ranging from 294 to 842 aa. All of them were localized in the cytoplasm or chloroplasts and unevenly distributed on the 11 chromosomes of G. pentaphyllum. The results of the phylogenetic tree showed that the G. pentaphyllum TPS gene family members could be divided into five subfamilies. As revealed by the analysis of promoter cis-acting elements, TPS gene family members in G. pentaphyllum were predicted to respond to a variety of abiotic stresses such as salt, low temperature, and dark stress. The analysis of gene expression patterns in different tissues of G. pentaphyllum revealed that nine TPS genes were tissue-specific in expression. The qPCR results showed that GpTPS16, GpTPS17, and GpTPS21 responded to a variety of abiotic stresses. This study is expected to provide references in guiding the further exploration of the biological functions of G. pentaphyllum TPS genes under abiotic stresses.

Progress on Pharmacological Effect of Terpinen-4-ol： A Review / 中国实验方剂学杂志

Terpine-4-ol is abundant in nature. As a cyclic monoterpenoid compound， terpine-4-ol is distributed in a variety of natural plants. It is the main component and the key active substance in many traditional Chinese essential oils， such as Melaleuca alba essential oil and coral ginger essential oil. Terpine-4-ol has anti-microbial， anti-tumor， insecticidal， anti-inflammatory， and other effects. It can treat cancer， as well as oral and cardiovascular diseases with great safety. In terms of antibacterial activity， terpine-4-ol can destroy bacterial cell walls， improve membrane permeability， and regulate bacterial migration， reproduction， and other related genes to inhibit bacterial activity. In terms of antifungal activity， terpine-4-ol can bind with ergosterol in fungal cell walls to cause fungal death. In terms of insecticidal activity， terpine-4-ol can inhibit Na+ and K+-ATPase activity and cause the death of the insect. In terms of anticancer activity， terpine-4-ol can regulate the expression of apoptosis-related proteins in cancer cells， so as to control the apoptosis of cancer cells. In this paper， the pharmacological activity and action mechanism of terpine-4-ol were reviewed to provide a reference for further research and utilization of terpine-4-ol.

Identification and analysis of terpene synthase (TPS) gene family in Schizonepeta tenuifolia / 中国中药杂志

Terpenoids are important secondary metabolites of plants that possess both pharmacological activity and economic value. Terpene synthases(TPSs) are key enzymes in the synthesis process of terpenoids. In order to investigate the TPS gene family members and their potential functions in Schizonepeta tenuifolia, this study conducted a systematic analysis of the TPS gene family of S. tenuifolia based on the whole genome data of S. tenuifolia using bioinformatics methods. The results revealed 57 StTPS members identified from the genome database of S. tenuifolia. The StTPS family members encoded 285-819 amino acids, with protein molecular weights ranging from 32.75 to 94.11 kDa, all of which were hydrophilic proteins. The StTPS family members were mainly distributed in the cytoplasm and chloroplasts, exhibiting a random and uneven physical localization pattern. Phylogenetic analysis showed that the StTPS genes family were divided into six subgroups, mainly belonging to the TPS-a and TPS-b subfamilies. Promoter analysis predicted that the TPS gene family members could respond to various stressors such as light, abscisic acid, and methyl jasmonate(MeJA). Transcriptome data analysis revealed that most of the TPS genes were expressed in the roots of S. tenuifolia, and qRT-PCR analysis was conducted on genes with high expression in leaves and low expression in roots. Through the analysis of the TPS gene family of S. tenuifolia, this study identified StTPS5, StTPS18, StTPS32, and StTPS45 as potential genes involved in sesquiterpene synthesis of S. tenuifolia. StTPS45 was cloned for the construction of an prokaryotic expression vector, providing a reference for further investigation of the function and role of the TPS gene family in sesquiterpene synthesis.

Avaliação da atividade antibacteriana do monoterpeno (r)-(+)-citronelal contra cepas de Pseudomonas aeruginosa

Introdução: Os monoterpenos, que são os principais constituintes da maioria dos óleos essenciais, compreendem uma série ampla de substâncias naturais e/ou sintéticas que já apresentaram diversas atividades biológicas. Objetivo: O presente estudo teve como objetivo avaliar a atividade antibacteriana do monoterpeno (R)-(+)-citro nelal contra cepas de Pseudomonas aeruginosa. Metodologia: Utilizou-se a técnica de diluição seriada para a determinação da concentração inibitória mínima (CIM) e também realizou-se a determinação da concentração bactericida mínima (CBM) do (R)-(+)-citronelal. Resultados: Os resultados obtidos para o monoterpeno testado foi de CIM igual à 512 µg/mL, apresentando assim um efeito antibacteriano forte e CBM variação de 512 a 1024 µg/mL, sendo assim considerado bactericida. Conclusão: Conclui-se que o monoterpeno (R)-(+)-citronelal devido ao seu potencial antimicrobiano pode ser considerado muito importante para o tratamento de várias enfermidades causadas pela Pseudomonas aeruginosa.

SUMMARY Introduction: Monoterpenes, which are the main constituents of most essential oils, comprise a wide range of natural and/or synthetic substances that have already exhibited diverse biological activities. Objective: The present study aimed to evaluate the antibacterial activity of monoterpene (R)-(+)-citronellal against Pseudomonas aeruginosa strains. Methodology: The serial dilution technique was used to determine the Minimum Inhibitory Concentration (MIC) and the Minimal Bactericidal Concentration (CBM) of (R)-(+)-citronellal was also determined. Results: The results obtained for the tested monoterpene were MIC50 equal to 512 µg/mL, thus presenting a strong antibacterial effect and CBM ranging from 512 to 1024 µg/mL, thus being considered bactericidal. Conclusion : Thus, it is concluded that the monoterpene (R)-(+)-citronellal, due to its antimicrobial potential, can be considered very important for the treatment of several diseases caused by Pseudomonas aeruginosa.

Introducción: Los monoterpenos, que son los componentes principales de la mayoría de los aceites esenciales, comprenden una amplia gama de sustancias naturales o sintéticas que ya han exhibido diversas actividades biológicas. Objetivo: El presente estudio tuvo como objetivo evaluar la actividad antibacteriana del monoterpeno (R)-(+)-citronelal contra cepas de Pseudomonas aeruginosa. Metodología: Se utilizó la técnica de dilución en serie para determinar la Concentración Inhibitoria Mínima (MIC) y también se determinó la concentración bactericida mínima (CBM) de (R)-(+)-citronelal. Resultados: Los resultados obtenidos para el monoterpeno ensayado fueron MIC50 igual a 512 µg/mL, presentando así un fuerte efecto antibacteriano y CBM que van desde 512 a 1024 µg/mL, por lo que se considera bactericida. Conclusión: así, se concluye que el monoterpeno (R)-(+)-citronelal, por su potencial antimicrobiano, puede considerarse muy importante para el tratamiento de diversas enfermedades causadas por Pseudomonas aeruginosa.

Transcriptome analysis to identify genes involved in the biosynthesis of aconitines in Aconitum pendulum / 药学学报

italic>Aconitum pendulum is a Tibetan medicine that is rich in bioactive compounds such as aconitine-type C19-diterpenoid alkaloids. To investigate the key enzymes in the aconitine biosynthesis pathway, roots, leaves and flowers of Aconitum pendulum were subjected to a high-throughput transcriptomic sequencing analysis by Illumina HiSeqTM2000. Trinity de novo assembly yielded 47 264 unigenes with an average length of 1 140 bp and N50 of 1 678 bp, of which 30 231 unigenes (63.96%) were annotated. In the KEGG database, 542 unigenes were implicated in 17 secondary metabolic pathways; the analysis showed that 44 genes encoded 20 key enzymes in the diterpene skeleton of aconitine biosynthesis and 12 BAHD acyltransferase genes were related to the acetylation modification, with differential expression among three organs. For example, ApTPS8 was the only committed enzyme in the upstream aconitine biosynthetic pathway. The high expression level of ApTPS8 in root indicated that it is the main tissue for the production of precursors of diterpene alkaloids. Consistent with the accumulation of aconitine, we propose that ApBAHD1/2/8 is involved in the biosynthesis of 2-hydroxyaconitine, dehydrated 14-benzoylaconitine, 8-O-methyl-14-benzoylaconine, benzoyldeoxyaconitine and benzoylaconitine, and ApBAHD10 is involved in the biosynthesis of acontine, lucidusculine, 14-O-acetylneoline and 14-O-acetylvirescenin. Comparative transcriptome analysis of A. pendulum and A. carmichaeli indicates significant gene loss in the family of diterpene synthases and acyltransferases in A. pendulum, which is in accordance with the significantly fewer type and quantity of aconitine compounds in this species. Therefore, A. pendulum has proved to be an ideal material for the study of the aconitine biosynthesis pathway. This work provides basic scientific data for further study of aconitine biosynthesis, the discussion of molecular mechanisms of toxicity, and the synthesis of genuine medicinal materials.

Quality Analysis and Suggestion for Ginkgo Folium Tablets and Ginkgo Folium Capsules / 中国实验方剂学杂志

Objective:To conduct quality evaluation of Ginkgo Folium preparations by analyzing the national evaluation sampling test results， analyze the quality differences， and put forward suggestions for the improvement of quality standards and market supervision. Method:The contents of total flavonol glycosides and terpene lactones in Ginkgo Folium tablets and Ginkgo Folium capsules were determined according to the methods of determination in the 2015 edition of Chinese Pharmacopoeia （the first volume）， and the contents of free flavonoids （quercetin， kaempferide and isorhamnetin） and sophoricoside in Ginkgo Folium preparations were determined according to related supplementary testing method of Ginkgo Folium tablets and Ginkgo Folium capsules issued by National Medical Products Administration. The quality differences of Ginkgo Folium preparations from different batches and different manufacturers were compared according to the contents of total flavonol glycosides， terpene lactones， free flavonoids and sophoricoside in 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules manufactured by 48 enterprises. Result:Quality of 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules was in accordance with the standard， but the contents of terpene lactones and total flavonol glycosides were all distributed in a wide range， and the quality of samples varied greatly among different enterprises. Conclusion:It is recommended that each enterprise should optimize the production process and strictly control the raw materials to ensure the consistency between different batches of samples.

Bioactive metabolites in of Ginkgo biloba leaves: variations by seasonal, meteorological and soil / Metabólitos bioativos em folhas de Ginkgo biloba: variações sazonais, meteorológicas e do solo

Abstract Ginkgo biloba is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. Variations of seasonal, meteorological and planting soil on the phytochemicals contents in G. biloba leaves due to the effects of growth meteorological and soil parameters were investigated in this study. The leaves of G. biloba were collected from different months and place in Zhejiang province, the contents of flavones (quercetin, kaempferol and isorhamnetin) and terpene lactones (bilobalide, ginkgolides A, B and C) were quantified by high performance liquid chromatography (HPLC) and the evaporative light scattering detector (ELSD) method. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Comprehensive analysis suggested the proper harvest time for G. biloba was in October of Zhejiang province. The result of correlation analysis with meteorological factors shows that the temperature and precipitation have non-significant effect on the main components of G. biloba. In addition, the type and content (Mn and Zn) of the soil showed significantly effect on the content of flavonoids and terpene lactones. This study enriched the knowledge on the development and utilization value of the G. biloba leaves and was useful for determining the optimal harvest time and growing condition.

Resumo Ginkgo biloba é um fitoterápico tradicional da medicina chinesa que contém vários componentes que contribuem para suas notáveis bioatividades. Variações sazonais, meteorológicos e de plantio do solo sobre os teores fitoquímicos em folhas de G. biloba, devido aos efeitos do crescimento meteorológico e parâmetros do solo, foram investigadas neste estudo. As folhas de G. biloba foram coletadas em diferentes meses e na província de Zhejiang, os teores de flavonas (quercetina, kaempferol e isorhamnetina) e lactonas terpênicas (bilobalida, ginkgolídeos A, B e C) foram quantificados por Cromatografia Líquida de Alta Eficiência (CLAE) e pelo método do detector de dispersão da luz por evaporação. Os métodos estabelecidos foram validados com boa linearidade, precisão, repetibilidade, estabilidade e recuperação. Uma análise abrangente sugeriu que o tempo de colheita adequado para G. biloba foi em outubro na província de Zhejiang. O resultado da análise de correlação com fatores meteorológicos mostra que a temperatura e a precipitação não têm efeito significativo sobre os principais componentes de G. biloba. Além disso, o tipo e composição (Mn e Zn) do solo apresentaram efeito significativo sobre o teor de flavonoides e lactonas terpênicas. Este estudo enriqueceu o conhecimento sobre o valor de desenvolvimento e utilização das folhas de G. biloba e foi útil para determinar o melhor tempo de colheita e condição de crescimento.

Cloning and analysis of a key enzyme of HMGR1 gene involved in terpene biosynthesis of Cornus officinalis / 中草药

Objective: To clone and analyze the cDNA sequence of 3-Hydroxy-3-methylglutaryl coenzyme-a reductase (HMGR1) of Cornus officinalis. Methods: In this study, specific primers were designed at both ends of the open reading frame (ORF) based on the unigene (c100572_g1) in the transcriptome data from C. officinalis. Subsequently, the cDNA sequence of CoHMGR1 gene was amplified by RT-PCR, cloned into the pTOPO-T vector and sequenced. This gene and its encoded protein were analyzed using bioinformatics methods. Results: The results suggested that CoHMGR1 was 2 116 bp in length andthe ORF was 1 338 bp in length, which encodes 445 amino acids and is a hydrophobic protein. Multiple sequence alignment and phylogenetic analysis showed that the amino acid sequence encoded by this gene has a high similarity with that of Camptotheca acuminate, reaching 79.56%. Based on the first comparison of transcriptome sequencing from leaves and fruits of C. officinalis, the CoHMGR1 gene was successfully cloned and analyzed. Conclusion: This study laid a foundation for studying the function of CoHMGR1 protein and the molecular mechanism of terpene biosynthesis pathway of C. officinalis.

Characterization of oil/water partition coefficient of chishao terpene glucoside components based on contribution rate of representative components for myocardial ischemia / 中国中药杂志

The components of traditional Chinese medicine(TCMCs) are the basic unit of raw materials for Chinese medicines, and their physical and chemical properties directly affect the choice of dosage forms and the optimization of prescriptions. However, most of TCMCs are multi-component complex systems, and the characterization of their overall properties is still in the exploration stage. On the basis of biological activity, the representative components are determined, and then the individual characteristics are fitted with the weight coefficient of efficacy contribution rate, which may provide reference for characterizing the overall properties of TCMCs. In this study, with the pharmacological effects of isoproterenol(ISO)-induced myocardial ischemia in rats as the indicators, the pharmacodynamic contribution rates of three representative components of chishao terpene glucoside components(CSTGCs) were evaluated by the normalization weighting method. The contribution rates of paeoniflorin, paeoniflorin and benzoylpaeoniflorin were 54.87%, 32.46% and 12.67%, respectively. The oil-water partition coefficients of paeoniflorin, albiflorin, benzoylpaeoniflorin in water and buffer solutions with different pH values were measured, and the oil-water partition coefficients of CSTGCs were characterized by the weight of their pharmacodynamics contribution rate. The results showed that the apparent oil-water partition coefficient(log P) of CSTGCs in the phosphate buffer system such as n-octanol-water(pH 2.0, 2.5, 5.0, 5.8, 6.8) were 0.18-0.22, indicating that CSTGCs have common absorption and low permeability, providing basis for the preparation of CSTGCs.

Comparative study of volatile components of Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts / 中草药

Objective To analyze the volatile components of Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts and compare the differences of these components in order to provide a theoretical basis for the usage of kernels of Magnolia Flos in classical famous prescriptions. Methods The volatile components were extracted by steam distillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The volatile components in different parts of Magnoliae Flos were identified by searching the retention index in the mass spectrometry database. The relative content of volatile components was calculated by peak area normalization method, and the data of GC-MS was further proceeded by principal component analysis (PCA). Results The average volatile components content in Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts was 1.70%, 6.79%, and 0.45%, respectively. The numbers of compounds identified by GC-MS in those three different parts were 87, 82, and 85, respectively. These compounds mainly included terpene olefins and alcohols type chemicals. PCA method was then applied to process the GC-MS data and distinguish different parts and habitats of Magnoliae Flos. Conclusion The volatile components in the kernels of Magnoliae Flos were generally the same as those of Magnoliea Flos, while the content of which was much higher than those of whole flower bud of Magnoliae Flos. The results indicated that it was rational to use the kernels of Magnolia Flos in classical famous prescriptions.

A new triterpenoid sapogenin from flowers of Bombax malabaricum / 中草药

Objective To study the chemical constituents from the flowers of Bombax malabaricum. Methods The compound was separated and purified by silica gel and gel column choromatography. The structure of the new compound was identified by NMR, HRMS (EIS), IR, UV, and single crystal X-ray diffraction. Results A new compound was isolated from the flowers of B. malabaricum and identified as 3β-acetoxy-22α,30-dihydroxyurs-20-ene. Conclusion Compound 1 was a ursane-type triterpenoid and named as mumian terpene A.

Discovery and verification of representative ingredients of Chishao terpene glucoside components against ischemia and hypoxia injury / 药学学报

The discovery and verification of components are prerequisites for developing of component preparations. The molecular docking technique and pharmacodynamic activity evaluation provide effective methods for the discovery and verification of the representative components of Chishao terpene glucoside components (CSTGCS) against ischemia and hypoxia injury. The chemical constituents of CSTGCS were analyzed qualitatively by UPLC-TOF/MS/MS. Main chemical constituents were docked with key receptor proteins of myocardial ischemia to preliminarily screen anti-ischemia active ingredients, and screening for main active ingredients with Libdockscore. Then a H9c2 cell hypoxia injury model was established, and creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase (SOD), malondialdehyde (MDA) were determined to screen the representative combinations in CSTGCS. In addition, apoptosis index, apoptotic protein expression and mitochondria-associated mRNA levels were determined to verify the inhibition of the representative components (RCS) on the apoptosis of hypoxic cells. Eventually, the representative components of CSTGCS were determined. The results showed that paeoniflorin, albiflorin, benzoyl paeoniflorin and oxypaeoniflorin were considered to be the main active components because of their high matching with target proteins (4TWT, 3O4O, 4KZN, 1M9J) in space and energy. There was no statistical difference in regulating CK, LDH, SOD, MDA levels and maintaining mitochondrial function as well as inhibiting cell apoptosis between CSTGCS group and RCS group (paeoniflorin + albiflorin + benzoyl paeoniflorin combination). Therefore, paeoniflorin, albiflorin and benzoyl paeoniflorin were selected as the most representative ingredients of CSTGCS against ischemia and hypoxia injury, providing a basis for the overall properties of the components and formulation of CSTGCS.

Correlation analysis between the chemical contents and bioactivity for the quality control of Alismatis Rhizoma

In order to clarify regions of production and to discriminate processing methods, quantitative and qualitative analyses for saccharides and terpenes in 35 batches of Alismatis Rhizoma were performed. Methodologies included HPLC-PDA, HPLC-VWD and UHPLC-MS , combined with principal component analysis (PCA) and partial least squares regression techniques (PLSR). The inhibitory effects of triterpenes and Alismatis Rhizoma extracts on lipase activity were evaluated . PLSR analysis revealed significant positive correlations ( = 0.5795) between the contents of triterpenes , , , and and the inhibitory effects of Alismatis Rhizoma. The present study establishes an effective method for simultaneous determination of multiple components, and identifies key bioactive triterpenes. These results can be used for systematic and novel analytical strategies for the quality control of Alismatis Rhizoma production.

Progress in biosynthesis of santalene and santalol / 生物工程学报

Santalene and santalol are the main components of valuable perfume sandalwood essential oil, and have good antibacterial, anti-oxidation and anti-tumor activities. Commercial sandalwood essential oil is mainly extracted from sandalwood tree that grows slowly and is difficult to cultivate. In addition, the extraction recovery of sandalwood essential oil from sandalwood tree is too low to meet the market demand. These factors make sandalwood essential oil expensive. An option is to use genetic engineering and molecular biological methods to heterologously express related synthase of santalene and santalol in microbial host. In this paper, the biosynthesis progress of santalene and santalol synthase, as well as the optimization of mevalonate metabolic pathways in the hosts are summarized. Furthermore, the strategies of applying protein engineering technology to carry out orthomutation of santalene synthase were also discussed, to provide reference for the optimal biosynthesis of santalene and santalol.

Research development of ginkgo terpene lactones / 中国中药杂志

Ginkgo terpene lactones, as an important active ingredient from Ginkgo leaves, has high medicinal values and has been widely used in clinics. This article would review the researches both at home and abroad, including chemical composition, structure-activity relationship, analytical methods, pharmacological effects, pharmacokinetic characteristics, and so on, providing a reference for further development and utilization of ginkgo terpene lactones.

Study on peel components and juice quality of three mandarin hybrids (Citrus reticulata) on sour orange rootstock cultivated in Ramsar, Iran / Estudio sobre los componentes de la cáscara y la calidad del jugo de tres híbridos de mandarina (Citrus reticulata) en patrones de naranja agria cultivados en Ramsar, Irán

The peel components and juice quality of three mandarin hybrids were investigated in this study. Peel components were extracted using the cold-press method and analyzed using GC-FID and GC-MS. Total soluble solids, total acid, pH value, ascorbic acid as well as density were determined in juice obtained from mandarin hybrids. Twenty-six, thirty-five and nineteen peel components were identified in Fortune, Robinson and Osceola respectively including: aldehydes, alcohols, esters, monoterpenes, sesquiterpenes and other components. The major components were limonene, γ-terpinene, (E)-ß-ocimene, ß-myrcene, sabinene, linalool and α-Pinene. Among the three scions examined, Fortune showed the highest content of aldehydes and Robinson showed the highest content of TSS. This study shows that scion has a profound influence on aldehyd and TSS that are important to quality improvement.

Los componentes de la cáscara y la calidad del jugo de tres híbridos de mandarina fueron investigados en este estudio. Los componentes de la cáscara fueron extraídos por el método de prensado en frío y se analizaron mediante GC-FID y GC-MS. el total de sólidos solubles, acidez total, pH, ácido ascórbico, así como la densidad se determinaron en el jugo obtenido a partir de híbridos de mandarina. Veintiséis, treinta y cinco y diecinueve componentes de la cáscara de Fortuna, Robinson y Osceola respectivamente fueron identificados, incluyendo: aldehídos, alcoholes, ésteres, monoterpenos, sesquiterpenos y otros componentes. Los componentes principales eran limoneno, γ-terpineno, (E) -ß-ocimeno, ß-mirceno, sabineno, linalool y α-pineno. Entre los tres vástagos examinados, Fortune mostró el mayor contenido de aldehídos y Robinson mostró el mayor contenido de SST. Este estudio muestra que el vástago tiene una profunda influencia en aldehído y SST que son importantes para mejorar la calidad.

Bioactive metabolites in of Ginkgo biloba leaves: variations by seasonal, meteorological and soil

Abstract Ginkgo biloba is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. Variations of seasonal, meteorological and planting soil on the phytochemicals contents in G. biloba leaves due to the effects of growth meteorological and soil parameters were investigated in this study. The leaves of G. biloba were collected from different months and place in Zhejiang province, the contents of flavones (quercetin, kaempferol and isorhamnetin) and terpene lactones (bilobalide, ginkgolides A, B and C) were quantified by high performance liquid chromatography (HPLC) and the evaporative light scattering detector (ELSD) method. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Comprehensive analysis suggested the proper harvest time for G. biloba was in October of Zhejiang province. The result of correlation analysis with meteorological factors shows that the temperature and precipitation have non-significant effect on the main components of G. biloba. In addition, the type and content (Mn and Zn) of the soil showed significantly effect on the content of flavonoids and terpene lactones. This study enriched the knowledge on the development and utilization value of the G. biloba leaves and was useful for determining the optimal harvest time and growing condition.

Resumo Ginkgo biloba é um fitoterápico tradicional da medicina chinesa que contém vários componentes que contribuem para suas notáveis bioatividades. Variações sazonais, meteorológicos e de plantio do solo sobre os teores fitoquímicos em folhas de G. biloba, devido aos efeitos do crescimento meteorológico e parâmetros do solo, foram investigadas neste estudo. As folhas de G. biloba foram coletadas em diferentes meses e na província de Zhejiang, os teores de flavonas (quercetina, kaempferol e isorhamnetina) e lactonas terpênicas (bilobalida, ginkgolídeos A, B e C) foram quantificados por Cromatografia Líquida de Alta Eficiência (CLAE) e pelo método do detector de dispersão da luz por evaporação. Os métodos estabelecidos foram validados com boa linearidade, precisão, repetibilidade, estabilidade e recuperação. Uma análise abrangente sugeriu que o tempo de colheita adequado para G. biloba foi em outubro na província de Zhejiang. O resultado da análise de correlação com fatores meteorológicos mostra que a temperatura e a precipitação não têm efeito significativo sobre os principais componentes de G. biloba. Além disso, o tipo e composição (Mn e Zn) do solo apresentaram efeito significativo sobre o teor de flavonoides e lactonas terpênicas. Este estudo enriqueceu o conhecimento sobre o valor de desenvolvimento e utilização das folhas de G. biloba e foi útil para determinar o melhor tempo de colheita e condição de crescimento.

Applications of Reference Extract from Ginkgo Leaf Total Lactones for Quantitative Assay of Ginkgo Folium / 医药导报

Objective To explore the feasibility of using the quantitative reference extract of ginkgo leaf total lactones instead of single component reference for the quantitative assay of Ginkgo Folium.Methods HPLC-ELSD method was performed by using a Diamonsil C18 column (250 mm×4.6 mm,5 μm) with methanol-water as the mobile phase at the gradient elution mode.Flow rate was 1.0 mL·min-1.The parameters of ELSD detector were as follows,the drifit tube temperature was 105 ℃,and the flow rate of nitrogen(N2) was 3 L·min-1.Results The linear ranges of ginkgolide A,ginkgolide B,ginkgolide C,and bilobalide were 0.735-5.879 μg (r=0.999 6),0.404-6.060 μg (r=0.999 6),0.296-4.439 μg (r=0.999 6),and 1.001-6.006 μg (r=0.999 7),respectively.The recoveries and RSD of the four components were 95.6% (4.0%),97.3% (4.5%),99.3% (5.0%),and 100.4% (2.1%),respectively.Conclusion The quantitative reference extract of ginkgo leaf total lactones can be used as the substitute for the determination of terpene lactones.