Zhisheng Xie

Persimmon (Diospyros kaki L.) leaves: a review on traditional uses, phytochemistry and pharmacological properties.

ETHNOPHARMACOLOGICAL RELEVANCE Persimmon (Diospyros kaki L.) leaves, known as Shi Ye (in Chinese), have a long history as a Chinese traditional medicine for the treatment of ischemia stroke, angina, internal hemorrhage, hypertension, atherosclerosis and some infectious diseases, etc. Additionally, persimmon leaves could be used as healthy products, cosmetics and so on, which have become increasingly popular in Asia, such as Japan, Korea and China etc. AIM OF THE REVIEW The present paper reviewed the ethnopharmacology, phytochemistry, analytical methods, biological activities and toxicology of persimmon leaves in order to assess the ethnopharmacological use and to explore therapeutic potentials and future opportunities for research. MATERIALS AND METHODS Information on persimmon leaves were gathered via the Internet (using Google Scholar, Baidu Scholar, Elsevier, ACS, Pudmed, Web of Science, CNKI and EMBASE) and libraries. Additionally, information was also obtained from some local books. RESULTS Persimmon leaves have played an important role in Chinese system of medicines. The main compositions of persimmon leaves were flavonoids, terpenoids, etc. Scientific studies on extracts and formulations revealed a wide range of pharmacological activities, such as, antioxidative, hypolipidemic, antidiabetic, antibacterial, hemostasis activities and effects on cardiovascular system. Based on the pharmacological activities, persimmon leaves were widely used in clinic including treatment of cardiovascular disease, hemostasis, antibacterial, anti-inflammatory and beauty treatment. CONCLUSIONS Persimmon leaves probably have therapeutic potential in the prevention and treatment for cerebral arteriosclerosis, diabetes, hypertension. It showed significant neuroprotection against ischemia/reperfusion injury in vivo and in vitro. Moreover, it can regulate immune function and inhibite inflammation. Further investigations are needed to explore individual bioactive compounds responsible for these pharmacological effects in vitro and in vivo and the mode of actions. Further safety assessments and clinical trials should be performed before it can be integrated into medicinal practices.

Extraction and isolation of flavonoid glycosides from Flos Sophorae Immaturus using ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography

A method of ultrasonic-assisted extraction followed by high-speed countercurrent chromatography was established for the extraction and isolation of three flavonoid glycosides, i.e. rutin, narcissin, and nicotiflorin from Flos Sophorae Immaturus. The effects of ultrasonic-assisted extraction factors for the main flavonoid compound (rutin) from Flos Sophorae Immaturus were optimized using Box-Behnken design combined with response surface methodology. The optimum conditions were determined as ultrasonic power 83% (600 W), solvent-to-material ratio 56:1, methanol concentration 82% v/v, and extraction time 60 min. Three bioactive flavonol glucosides, rutin, narcissin, and nicotiflorin were isolated from Flos Sophorae Immaturus using high-speed countercurrent chromatography. The separation was performed with a two-phase solvent system containing ethyl acetate/n-butanol/methanol/water (4:0.9:0.2:5, v/v). Amounts of 87 mg of rutin, 10.8 mg of narcissin, and 1.8 mg of nicotiflorin were isolated from 302 mg of crude extract of Flos Sophorae Immaturus in a one-step separation within 160 min with purities of 99.3, 98.0, and 95.1%, respectively, as determined by HPLC with diode array detection. Their structures were characterized by UV, MS, and NMR spectroscopy. It was demonstrated that the established method was simple, fast, and convenient, which was feasible to extract and isolate active flavonoid glycosides from Flos Sophorae Immaturus.

Chemical fingerprint and simultaneous determination of flavonoids in Flos Sophorae Immaturus by HPLC-DAD and HPLC-DAD-ESI-MS/MS combined with chemometrics analysis

A simple and efficient HPLC method is developed for the fingerprint analysis of Flos Sophorae Immaturus and for the simultaneous determination of three flavonoids in Flos Sophorae Immaturus, namely rutin, narcissin and quercetin. The separation of analytes is conducted on a Dikma Diamonsil C18 column (250 mm × 4.6 mm, 5 μm) at 35 °C; the wavelength of UV detector was set at 254 nm; the mobile phases were composed of acetonitrile and aqueous acetic acid (0.5%, v/v). A gradient elution was carried out at a flow rate of 1.0 mL min−1. A common chromatographic fingerprint consisting of 9 characteristic peaks is established among 14 batches of Flos Sophorae Immaturus samples, which were collected from different source areas of China. The constituents in Flos Sophorae Immaturus were further identified by HPLC-DAD-ESI-MS/MS and most of them were flavonoids. Multiple chemometrics analysis, including similarity analysis (SA), hierarchical cluster analysis (HCA) and principal component analysis (PCA) were performed to classify samples. In quantitative analysis, all of the calibration curves showed good linear regression (R2 ≥ 0.9991) within the tested ranges, and the mean recoveries ranged from 100.28% to 101.08%. Three flavonoid compounds in Flos Sophorae Immaturus were simultaneously quantified by the established method. The results demonstrate that the developed method is accurate and effective, and could be readily utilized for the comprehensive quality control of Flos Sophorae Immaturus.

Volatile components of Rhizoma Alpiniae Officinarum using three different extraction methods combined with gas chromatography–mass spectrometry

Volatile components from Rhizoma Alpiniae Officinarum were respectively extracted by three methods including hydrodistillation, headspace solid-phase microextraction (HS-SPME) and diethyl ether extraction. A total of 40 (hydrodistillation), 32 (HS-SPME) and 37 (diethyl ether extraction) compounds were respectively identified by gas chromatography–mass spectrometry (GC/MS) and 22 compounds were overlapped, including α-farnesene, γ-muurolene, 2,6-dimethyl-6-(4-methyl-3-pentenyl)bicyclo[3.1.1]hept-2-ene, eucalyptol and cadina-1(10), 4-diene and so forth, varying in relative contents. HS-SPME is fast, sample saving and solvent-free and it also can achieve similar profiles as those from hydrodistillation and solvent extraction. Therefore, it can be the priority for extracting volatile components from medicinal plants.

The GC/MS Analysis of Volatile Components Extracted by Different Methods from Exocarpium Citri Grandis.

Volatile components from Exocarpium Citri Grandis (ECG) were, respectively, extracted by three methods, that is, steam distillation (SD), headspace solid-phase microextraction (HS-SPME), and solvent extraction (SE). A total of 81 compounds were identified by gas chromatography-mass spectrometry including 77 (SD), 56 (HS-SPME), and 48 (SE) compounds, respectively. Despite of the extraction method, terpenes (39.98~57.81%) were the main volatile components of ECG, mainly germacrene-D, limonene, 2,6,8,10,14-hexadecapentaene, 2,6,11,15-tetramethyl-, (E,E,E)-, and trans-caryophyllene. Comparison was made among the three methods in terms of extraction profile and property. SD relatively gave an entire profile of volatile in ECG by long-time extraction; SE enabled the analysis of low volatility and high molecular weight compounds but lost some volatiles components; HS-SPME generated satisfactory extraction efficiency and gave similar results to those of SD at analytical level when consuming less sample amount, shorter extraction time, and simpler procedure. Although SD and SE were treated as traditionally preparative extractive techniques for volatiles in both small batches and large scale, HS-SPME coupled with GC/MS could be useful and appropriative for the rapid extraction and qualitative analysis of volatile components from medicinal plants at analytical level.

Optimization of the fractional precipitation of paclitaxel from a Taxus chinensis cell culture using response surface methodology and its isolation by consecutive high‐speed countercurrent chromatography

A consecutive preparation method for the isolation and purification of paclitaxel from the Taxus Chinensis cell culture was developed in this study. The process involved alkaline Al2O3 chromatography, fractional precipitation, and high-speed countercurrent chromatography. The original cell culture materials were first extracted with methanol using ultrasound-assisted extraction, and then the extract (the content of paclitaxel is 1.5%) was separated by alkaline Al2O3 column chromatography. Subsequently, fractional precipitation was used to obtain paclitaxel. In particular, response surface methodology was used to optimize the factors of fractional precipitation (methanol concentration, material-to-solvent ratio, and precipitating time were optimized as 48.14%, 8.85 mg/mL, and 48.71 h, respectively) and the yield of fractional precipitation product was 30.64 ± 0.60 mg (the content of paclitaxel is 89.3%, 27.37 ± 0.54 mg) from a 100 mg fraction by Al2O3 column separation (the content of paclitaxel is 32.4%). Then, the product was used for further isolation by high-speed countercurrent chromatography. About 1.00 g paclitaxel (200 ± 2 mg in each loading) with a purity up to 99.61% was isolated from 1.25 g of fractional precipitation product with a solvent system of n-hexane/ethyl acetate/methanol/water (1.2:1.8:1.5:1.5, v/v/v/v) in one run of five consecutive sample loadings without exchanging a new solvent system.

Preparative isolation of paclitaxel and related taxanes from cell cultures of Taxus chinensis using reversed-phase flash chromatography.

In this study, paclitaxel, baccatin III, taxuyunnanine C and sinenxane C were successfully separated by reversed-phase flash chromatography on a manually packed C18 column from Taxus chinensis cell culture extract. The crude cell culture extract was first treated with Al2O3 column chromatography and then divided into two parts: fraction 1 and fraction 2. Ten milligrams of baccatin III and 19 mg of paclitaxel were obtained from 100 mg dried fraction 1. Fifty-two milligrams of taxuyunnanine C and 11 mg sinenxane C were obtained from 100 mg dried fraction 2. The purities of the four compounds were 98.02%, 98.53%, 98.93% and 98.76%, respectively. Their structures were characterised by using UV, MS and NMR. These results indicate that paclitaxel and related taxanes including baccatin III can be obtained from cell culture in a highly pure state using reversed-phase flash chromatography.

ISOLATION AND PURIFICATION OF PAEONIFLORIN AND ALBIFLORIN FROM RADIX PAEONIAE RUBRA BY HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

Paeoniflorin and albiflorin were isolated and purified from Radix Paeoniae Rubra by high-speed counter-current chromatography with a solvent system of ethyl acetate-n-butanol-water (3:2.5:5, v/v/v) in one step. From 100 mg of the n-butanol extract of Radix Paeoniae Rubra, 10 mg of paeoniflorin and 4.3 mg of albiflorin were obtained with purity of 98.75% and 95.03%, as determined by HPLC. Their structures were identified by MS, UV, and NMR analysis. In this study, a rapid method for isolation and purification of the two major compounds from Radix Paeoniae Rubra crude extract was established.

Isolation and Purification of Geniposide, Crocin-1, and Geniposidic Acid from the Fruit of Gardenia jasminoides Ellis by High-Speed Counter-Current Chromatography

Geniposide, crocin-1, and geniposidic acid were simultaneously separated from the fruit of Gardenia jasminoides Ellis by one step of high-speed counter-current chromatography with the solvent system composed of ethyl acetate-n-butanol-water(1:4:5, v/v/v) within 130 min. The purities of the three compounds were 98.7%, 97.1%, and 90.4%, respectively, as determined by HPLC. Their structures were confirmed by MS, UV, 1H NMR, and13C NMR analysis.

ISOLATION AND PURIFICATION OF 20-HYDROXYECDYSONE FROM RADIX SERRATULAE CHINENSIS BY HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

□ The bioactive compound 20-Hydroxyecdysone was successfully isolated and purified from Radix Serratulae Chinensis by high-speed counter-current chromatography with a solvent system of ethyl acetate–n-butanol–water (4:1:5, v/v/v) in one step. From 230mg of the n-butanol extract of Radix Serratulae Chinensis, 16.7mg of 20-Hydroxyecdysone was obtained with purity of 99.3%, as determined by high performance liquid Chromatography (HPLC). The structure was identified by mass spectrophotometry (MS), ultraviolet spectrophotometry (UV), and nuclear magnetic resonance spectrophotometry (NMR) analysis. In this study, a rapid method for isolation and purification of the major compound from Radix Serratulae Chinensis extract was established.