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.

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.

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.

Establishment of fingerprint of Ginkgo biloba leaves and study on its quality evaluation / 中草药

To compare the quality Ginkgo biloba L. leaves collected form various habitats, different tree ages, and different harvesting times and to confirm the source of G. biloba leave used in Xin Mai Capsule. To establish the HPLC fingerprint of G. biloba leaves to determine the fingerprint and content of total flavonol glucoside and terpene lactones in G. biloba leaves collected from various habitats, different tree ages, and different harvesting time, and then to analyze the results. The contents of total flavonol glucoside and terpene lactones and fingerprint similarity in G. biloba leaves from various regions, different tree ages, and collecting times were different. The fingerprint control method has provided the scientific basis for the reasonable collection and quality evaluation of G. biloba leaves. The G. biloba leaves in June to August, 2-3 year-old from Pizhou, Jiangsu province are better, which could be considered as the source of G. biloba leaves used in Xin Mai Capsule.

Comprehensive analysis of chemical constituents in Xingxiong injection by high performance liquid chromatography coupled with mass spectrometry / 中国天然药物

Xingxiong injection (XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacological studies, chemical analysis and quality control studies on this formula are relatively limited. In the present study, a high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in XXI. According to the fragmentation rules and previous reports, thirty ginkgo flavonoids, four ginkgo terpene lactones, and one alkaloid were identified. A high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QQQ MS) method was then applied to quantify ten major constituents in XXI. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. The total contents of ginkgo flavonoids were about 22.05-25.51 μg·mL(-1) and the ginkgo terpene lactones amounts were about 4.41-8.70 μg·mL(-1) in six batches of XXI samples, respectively. Furthermore, cosine ratio algorithm and distance measurements were employed to evaluate the similarity of XXI samples, and the results demonstrated a high-quality consistency. This work could provide comprehensive information on the quality control of Xingxiong injection, which be helpful in the establishment of a rational quality control standard.

Orthogonal Experiment Study on Extraction Process of Ginkgo Leaves / 中国中医药信息杂志

Objective To determine the best extraction process of ginkgo leaves with the total transfer rate of total flavonol glycosides and terpene lactones as the index.Methods The effects of ethanol concentration, solid-liquid ratio and extraction time on extraction process were investigated by orthogonal design method, and the contents of total flavonol glycosides and terpene lactones were detected by HPLC to calculate transfer rate.Results The optimum extraction conditions were as follows:85% ethanol refluxing and extracting for three times;the first time extracting with five-fold amount of solvent (V/W) for 3 hours;the last two times with three-fold solvent (V/W) for 2 hours.Conclusion This extraction process has the advantages of simplicity of operator, reason, energy conservation, high efficiency, and is suitable for industrial production.

Fingerprint Analysis of Terpene Lactones in Ginkgo Biloba Tablets by HPLC-ELSD Coupled with Chemometrics / 世界科学技术-中医药现代化

This study was aimed to establish a high performance liquid chromatography (HPLC) method coupled with Evaporative Light-scattering Detector (ELSD) in order to develop the determination of fingerprint of terpene lactones in Ginkgo biloba tablets. An Agilent Extend-C18 (4.6 mm í 250 mm, 5 μm) was employed as the analysis column and the normal propyl alcohol-tetrahydrofuran-water (1:15:84) as mobile phase. The column temperature was 30℃. And the flow rate was 1.0 mL·min-1. HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) was used to identify the common peaks. The fingerprint was further evaluated by chemometrics methods including principal component analysis (PCA), similarity analysis (SA) and hierarchical clus-tering analysis (HCA). The results showed that the precision, stability and repeatability of this method were favorable. Five common peaks were identified by LC/Q-TOF MS as ginkgolide J ( M ) , C , A , B and bilobalide , respectively . Fourteen batches of Ginkgo biloba tablets were determined. With the aid of PCA, SA and HCA, the common pattern of the fingerprint of terpene lactones was established. Samples were divided into 4 clusters by their quality differ-ence. It was concluded that the method established in this paper can be used for quality evaluation of terpene lac-tones in G ink go b ilob a tablets.

Study on changing rules for total flavonoids and total terpene lactones in Ginkgo biloba leaves at different ages of tree and collecting seasons / 中草药

Object To define suitable gathering season of Ginkgo biloba L. leaves. Methods The content of total flavonoids and total terpene lactones in G. biloba leaves were determined by HPLC DAD and HPLC ELSD. Results The content of total flavonoids and total terpene lactones in G. biloba were distinct in different ages of tree and collecting seasons. Conclusion The content of total flavonoids and total terpene lactones in G. biloba are the highest in 2 3 ages of tree. The total flavonoid is the highest in May and total terpene lactone is the highest in September.

Quality and quantity analysis of terpene lactones from Ginkgo biloba leaves by GC-MS / 中草药

Object To develop a capillary GC-MS analytical method for identification and deter- mination of ginkgolide A, B, C and bilobalide (GA, GB, GC and BB) in Ginkgo biloba L. leaves. Methods The leave samples were extracted in ultrasonic bath with ethanol-water (20∶80). The extract was purified by liquid-liquid extraction with ethyl acetate followed by solid-phase extraction on a column mixed with acid Al 2O 3, active carbon and celite. The terpenes were trimethylsilylated by BSTFA (with 1% TMCS) for 60 min at 100 ℃ and determined by GC-MS with HP-5 MS capillary column in the selected-ion monitoring mode. The intense fragment ions were chosen as monitoring ions for quantitative analysis. Cholesterol was used as an internal standard. Column temperature gradient: initial temperature 180 ℃, maintained 1 min, and then increased at 20 ℃/min to 260 ℃, and finally at 2 ℃/min up to 300 ℃, maintained 2 min. Results The retention times of GA, GB, GC and BB were 13.7,14.3,15.3 and 6.8 min, the major fragmentation ions (monitoring) were at m/z 537, 625, 713 and 455 (299), the average recoveries of GA, GB, GC and BB were 102.0%, 99.4%, 96.0%, 96.3%, RSD were 0.54%, 2.40%, 1.98% and 2.43%, respectively. Conclusion This method is repeatable, specific, accurate and easy to operate. It is adoptable for quality and quantity analysis of terpene lactones from G. biloba leaves.

Quality analysis of seedling leaves of Ginkgo biloba Ⅱ / 中草药

Object To stuay the quality of seedling leaves cultured from improved variety of Ginkgo biloba L.. Methods The content of terpene lactones in seedling leaves of G. biloba were determined by RP-HPLC-ELSD.Results The content of total terpene lactone producted in Jun. was more than that in Aug., and it was obviously lower in Nov.. The content of terpene lactone in grafting was more than that in seedling. The content of terpene lactone in male trees was more than that in female trees.Conclusion The content of the active components was related to the season of growth, age of tree, raising modes, environment of growth, geographic conditions and others. The evaluation of the content of the main active components BB and GB showed that the quality of seedling leaves cultured from improved variety of G. biloba in Jinagxi is better than that reported by literature.

Study on the development variation of the content of terpene lactones of Ginkgo Biliba Leaves / 中成药

Objective: To study the effectc of different growing areas and seasonal variation on the components and contents of Ginkgo Biloba terpene lactones. Methods: The contents and components of different Ginkgo Biloba terpene lactones were determined by RP-HPLC. Results: The contents of Ginkgo Biloba terpene lactones among the samples of Ginkgo Biloba Leaves of different growing areas were obviously different, the terpene lactones content of Ginkgo Biloba Leaves growing in Zaoyang county, Hubei Province, was higher. Based on the monthly variation of the contents of different Ginkgo Biloba terpene lactones, the suitable seasons for the harvest of Ginkgo Biloba Leaves were from Sep. to Oct. Conclusion: The growing areas and seasonal variation had great influence over the contents and components of Ginkgo Biloba terpene lactones.