Screening for the extracorporeal coagulation activity quality markers (Q-markers) of dried and stir fried ginger

Ginger is a common condiment that is widely used as Chinese medicine in China and Southeast Asia. Dried ginger and stir fried ginger are two common processed products of ginger, with distinct clinical uses. The aim of this study was to identify the chemical components (quality markers, Q-mark) responsible for the differences in in vitro hemostatic activity between dried and stir fried ginger and to provide a basis for the selection between the two types of ginger in clinical application. In this study, methanolic extracts of dried and stir fried ginger were characterized using UPLC-Q/TOF-MS and then evaluated for in vitro coagulation activity. Spectral effect correlation analysis was used to identify quality markers, while molecular docking simulation was used to evaluate the binding energy between potential active compounds and target proteins. A total of 49 chemical constituents of the ginger extracts were identified using UPLC-Q/TOF-MS, 27 of which were significantly different between the two extracts. A fingerprint of 18 batches of dried and stir fried ginger established that zingiberone, 6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol, 8-shogaol, and 10-shogaol were common constituents of the two extracts. Results of coagulation assays revealed that dried ginger had anti-coagulation effects, while stir fried ginger had hemostatic effect. Zingiberone, 6-shogaol and 10-shogaol were identified as the active components responsible for the hemostatic effect of Stir fried ginger through multivariate statistical analysis. In addition, molecular docking simulations suggested that these three components bound to Src proteins on platelets. Consequently, 6-gingerol, zingiberone, 6-shogaol and 10-shogaol were selected as quality markers to distinguish between dried and stir fried ginger. These results provide scientific evidence for the establishment of a quality evaluation system for the integrity and specificity of dried and stir fried ginger.     

超高效液相色谱-串联质谱法快速同时测定生姜中10种营养成分

建立了超高效液相色谱-串联质谱（UHPLC-MS/MS）快速同时测定生姜中姜辣素类和姜黄素类营养成分的分析方法，具体包括6-姜酚、8-姜酚、10-姜酚、6-姜烯酚、8-姜烯酚、10-姜烯酚、四氢姜黄素、姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素10种目标物。采用ZORBAX RRHD Eclipse Plus C18色谱柱（100 mm×2.1 mm，1.8 μm）分离，以0.1%（v/v）甲酸水溶液和0.1%（v/v）甲酸甲醇溶液为流动相进行梯度洗脱，采用电喷雾电离（ESI）源、正离子和多反应监测（MRM）模式对目标物进行定性确证和定量分析。10种营养成分的线性相关系数（r）均≥ 0.9995，方法的定量限为0.10~7.71 μg/L，样品基质在3个水平下的平均加标回收率为82.8%~115.3%，相对标准偏差（RSD）为0.58%~11.49%。分析结果显示，生姜中10种营养成分均有检出，其中6-姜酚的含量最高且集中分布于373.35~702.48 mg/kg。该法简便快速，准确可靠，适用于生姜中姜辣素类和姜黄素类营养成分的分析，可为生姜质量鉴定和控制提供技术手段。     

Gastrointestinal motility enhancing effect of ginger and its active constituents

The effect of ginger root (Zingiberis Rhizoma) on gastrointestinal motility was examined based on its ability to enhance charcoal meal transport in mice. Oral administrations of the acetone extract of ginger (which contains volatile oils and bitter substances) at 75 mg/kg, [6]-shogaol at 2.5 mg/kg, or a [6]-, [8]- or [10]-gingerol at 5 mg/kg enhanced the transport of a charcoal meal. The effects of these substances were similar to or slightly weaker than those of metoclopramide and donperidone.     

Preparation of the monomers of gingerols and 6-shogaol by flash high speed counter-current chromatography

The flash high speed counter-current chromatographic (FHSCCC) separation of gingerols and 6-shogaol was performed on a HSCCC instrument equipped with a 1200-ml column (5 mm tubing i.d.) at a flow rate of 25 ml/min. The performance met the FHSCCC feature that the flow rate of mobile phase (ml) is equal to or greater than the square of the diameter of the column tubing (mm). The separation employed the upper phase of stationary phase of the n-hexane-ethyl acetate-methanol-water (3:2:2:3, v/v) as the stationary phase. A stepwise elution was performed by eluting with the lower phase of n-hexane-ethyl acetate-methanol-water (3:2:2:3, v/v) for first 90 min and the lower phase of the n-hexane-ethyl acetate-methanol-water (3:2:6:5, v/v) for the second 90 min. In each separation 5 g of the ethyl acetate extract of rhizomes of ginger was loaded, yielding 1.96 g of 6-gingerol (98.3%), 0.33 g of 8-gingerol (97.8%), 0.64 g of 6-shogaol (98.8%) and 0.57 g of 10-gingerol (98.2%). The separation can be expected to scale up to industrial separation.     

Ayurvedic preparation of Zingiber officinale Roscoe: effects on cardiac and on smooth muscle parameters

The rhizome of the Zingiber officinale Roscoe, a biennial herb growing in South Asia, is commonly known as ginger. Ginger is used in clinical disorders, such as constipation, dyspepsia, diarrhoea, nausea and vomiting and its use is also recommended by the traditional medicine for cardiopathy, high blood pressure, palpitations and as a vasodilator to improve the circulation. The decoction of ginger rhizome is widely used in Ayurvedic medicine. In this papery by high-performance liquid chromatography, we have seen that its main phytomarkers were 6-gingerol, 8-gingerol and 6-shogaol and we report the effects of the decoction of ginger rhizome on cardiovascular parameters and on vascular and intestinal smooth muscle. In our experimental models, the decoction of ginger shows weak negative inotropic and chronotropic intrinsic activities but a significant intrinsic activity on smooth muscle with a potency on ileum is greater than on aorta: EC₅₀ = 0.66 mg/mL versus EC₅₀ = 1.45 mg/mL.     

6-shogaol-rich extract from ginger up-regulates the antioxidant defense systems in cells and mice

The rhizome of ginger (Zingiber officinale Roscoe) is known to have several bioactive compounds including gingerols and shogaols which possess beneficial health properties such as anti-inflammatory and chemopreventive effects. Based on recent observations that 6-shogaol may have more potent bioactivity than 6-gingerol, we obtained a 6-shogaol-rich extract from ginger and examined its effects on the nuclear factor E2-related factor2 (Nrf2)/antioxidant response element (ARE) pathway in vitro and in vivo. 6-Shogaol-rich extract was produced by extracting ginger powder with 95% ethanol at 80 °C after drying at 80 °C (GEE8080). GEE8080 contained over 6-fold more 6-shogaol compared to the room temperature extract (GEE80RT). In HepG2 cells, GEE8080 displayed much stronger inductions of ARE-reporter gene activity and Nrf2 expression than GEE80RT. GEE8080 stimulated phosphorylations of mitogen-activated protein kinases (MAPKs) such as ERK, JNK, and p38. Moreover, the GEE8080-induced expressions of Nrf2 and HO-1 were attenuated by treatments of SB202190 (a p38 specific inhibitor) and LY294002 (an Akt specific inhibitor). In a mouse model, the GEE8080 decreased the diethylnitrosamine (DEN)-mediated elevations of serum aspartate transaminase and alanine transaminase as well as the DEN-induced hepatic lipid peroxidation. Inductions of Nrf2 and HO-1 by GEE8080 were also confirmed in the mice. In addition, the administration of GEE8080 to the mice also restored the DEN-reduced activity and protein expression of hepatic antioxidant enzymes such as superoxide dismutase, glutathione peroxidase and catalase. In conclusion, GEE8080, a 6-shogaol-rich ginger extract, may enhance antioxidant defense mechanism through the induction of Nrf2 and HO-1 regulated by p38 MAPK and PI3k/Akt pathway in vitro and in vivo.     

An efficient strategy for large-scale preparation of low polarity gingerols directly from ginger crude extract by high-speed countercurrent chromatography with different rotation mode

This study presents an efficient strategy for large-scale preparation of low polarity gingerols directly from ginger crude extract by high-speed countercurrent chromatography with different rotation mode. The ultrasonic-assisted extraction conditions were optimized by response surface methodology and the results showed the major low polarity gingerols could be well enriched under the optimized extraction conditions. Then the crude extract without any pretreatment was directly separated by high-speed countercurrent chromatography with different rotation mode using n-hexane/ethyl acetate/methanol/water (6:4:6:4, v/v/v/v) as the solvent system. In about 400 min, five major gingerols including 150 mg of [6]-gingerol, 50 mg of [8]-gingerol, 20 mg of [6]-shogaol, 43 mg of [6]-dehydrogingerdione, and 40 mg of [10]-gingerol were obtained from 1.2 g of crude extract in a single run with repeated injection. Their structures were identified by ¹H-NMR spectroscopy.     

Determination of 6-gingerol in ginger (Zingiber officinale) using high-performance thin-layer chromatography

A sensitive and accurate High-Performance TLC (HPTLC) method has been developed to determine the quantity of 6-gingerol in rhizomes of Zingiber officinale (family: Zingiberaceae), commonly known as ginger. Methanol extracts of rhizomes from three different sources were used for HPTLC, n-hexane, and diethyl ether (40:60 v/v) as the mobile phase. The Rf of 6-gingerol was found to be 0.40. The calibration plot was linear in the range of 250-1200 ng of 6-gingerol and the correlation coefficient of 0.9997 was indicative of good linear dependence of peak area on concentration. The mean quantity of 6-gingerol was found to be 60.44+/-2.53 mg/g of ginger extract. The method permits reliable quantification of 6-gingerol and good resolution and separation of 6-gingerol from other constituents of ginger. To study the accuracy and precision of the method, recovery studies were performed by the method of standard addition. Recovery values from 99.79 to 99.84% showed the excellent reliability and reproducibility of the method. The proposed HPTLC method for quantitative monitoring of 6-gingerol in ginger can be used for routine quality testing of ginger extracts.     

Liquid chromatographic determination of honokiol and magnolol in hou po (Magnolia officinalis) as the raw herb and dried aqueous extract

A validated analytical method is described for the determination of honokiol and magnolol in Hou Po (Magnolia officinalis) as the dried raw herb and the commercially prepared dried aqueous extract. The samples were extracted with methanol by the Soxhlet method, and the extract was analyzed by liquid chromatography with photodiode array (LC/PDA) detection with confirmation of analyte identity by negative-ion electrospray ionization tandem mass spectrometry (ESI-MS/MS). A C18 column was used with a menthanol--0.1% aqueous acetic acid gradient mobile phase. Honokiol and magnolol were quantified at 288 nm. With the MS detector, the honokiol precursor ion at m/z 265 was shown to produce ions at m/z 222 and 224. For magnolol, the precursor ion at m/z 265 produced the ions at m/z 247 and 245. Comparable results were obtained for the LC/PDA and LC/ESI-MS/MS methods of quantitation. Six commercially prepared dried aqueous extracts were analyzed. The levels of honokiol and magnolol found in the raw herb were 17.0 and 21.3 mg/g, respectively. The limits of detection for honokiol and magnolol in the raw herb were 0.45 and 0.58 mg/g, respectively, and in the dried aqueous extract, 0.04 and 0.30 mg/g, respectively.     

6-Gingesulfonic acid, a new anti-ulcer principle, and gingerglycolipids A, B, and C, three new monoacyldigalactosylglycerols, from zingiberis rhizoma originating in Taiwan.

By monitoring the effects on HCl/ethanol-induced gastric lesions in rats, a new antiulcer principle named 6-gingesulfonic acid was isolated from Zingiberis Rhizoma, the dried rhizome of Zingiber officinale Roscoe (cultivated and processed in Taiwan) together with three new monoacyldigalactosylglycerols named gingerglycolipids A, B and C. Their chemical structures were elucidated on the basis of chemical and physicochemical evidence. 6-Gingesulfonic acid showed more potent anti-ulcer activity than 6-gingerol and 6-shogaol.     

气相色谱-质谱技术分析姜油树脂中的挥发性及非挥发性成分

用超临界CO2萃取生姜根茎中的姜油树脂,并用气相色谱-质谱联用技术对其进行了成分分析。从姜油树脂中分析出77种化合物,其中挥发油成分50种,主要是α-姜烯(22.29%)、 β-倍半水芹烯(8.58%)、α-法尼烯(3.93%)、 β-没药烯(3.87%)和α-姜黄烯(2.63%)等倍半萜类化合物;姜辣素成分27种,主要成分为6-姜酚(9.38%)、6-姜烯酚(7.59%)和分析过程中由姜酚类或姜烯酚类化合物受热分解而形成的姜油酮(9.24%)。在姜辣素成分中,6-异姜酚、(Z)-10-异姜烯酚和(E)-10-异姜烯酚3种化合物是新发现的未见报道的化合物。实验中对这3种新化合物进行了质谱裂解分析。     

Anti-Platelet Aggregation and Vasorelaxing Effects of the Constituents of the Rhizomes of<em> Zingiber officinale</em>

In the present study, the chemical investigation of the bioactive fractions of the rhizomes of Zingiber officinale has resulted in the identification of twenty-nine compounds including one new compound, O-methyldehydrogingerol (1). Some of the isolates were subjected into the evaluation of their antiplatelet aggregation and vasorelaxing bioactivities. Among the tested compounds, [6]-gingerol (13) and [6]-shogaol (17) exhibited potent anti-platelet aggregation bioactivity. In addition, [10]-gingerol (15) inhibited the Ca²⁺-dependent contractions in high K⁺ medium. According to the results in the present research, the bioactivity of ginger could be related to the anti-platelet aggregation and vasorelaxing mechanism.     

Protective Effects of 6-Shogaol,an Active Compound of Ginger,in a Murine Model of Cisplatin-Induced Acute Kidney Injury

Acute kidney injury (AKI) is a dose-limiting side effect of cisplatin therapy in cancer patients. However, effective therapies for cisplatin-induced AKI are not available. Oxidative stress, tubular cell death, and inflammation are known to be the major pathological processes of the disease. 6-Shogaol is a major component of ginger and exhibits anti-oxidative and anti-inflammatory effects. Accumulating evidence suggest that 6-shogaol may serve as a potential therapeutic agent for various inflammatory diseases. However, whether 6-shogaol exerts a protective effect on cisplatin-induced renal side effect has not yet been determined. The aim of this study was to evaluate the effect of 6-shogaol on cisplatin-induced AKI and to investigate its underlying mechanisms. An administration of 6-shogaol after cisplatin treatment ameliorated renal dysfunction and tubular injury, as shown by a reduction in serum levels of creatinine and blood urea nitrogen and an improvement in histological abnormalities. Mechanistically, 6-shogaol attenuated cisplatin-induced oxidative stress and modulated the renal expression of prooxidant and antioxidant enzymes. Apoptosis and necroptosis induced by cisplatin were also suppressed by 6-shogaol. Moreover, 6-shogaol inhibited cisplatin-induced cytokine production and immune cell infiltration. These results suggest that 6-shogaol exhibits therapeutic effects against cisplatin-induced AKI via the suppression of oxidative stress, tubular cell death, and inflammation.     

Synergistic Anti-Inflammatory Activity of Ginger and Turmeric Extracts in Inhibiting Lipopolysaccharide and Interferon-γ-Induced Proinflammatory Mediators

This study aims to investigate the combined anti-inflammatory activity of ginger and turmeric extracts. By comparing the activities of individual and combined extracts in lipopolysaccharide and interferon-γ-induced murine RAW 264.7 cells, we demonstrated that ginger-turmeric combination was optimal at a specific ratio (5:2, w/w) in inhibiting nitric oxide, tumour necrosis factor and interleukin 6 with synergistic interaction (combination index < 1). The synergistic inhibitory effect on TNF was confirmed in human monocyte THP-1 cells. Ginger-turmeric combination (5:2, w/w) also upregulated nuclear factor erythroid 2–related factor 2 activity and heme oxygenase-1 protein expression. Additionally, 6-shogaol, 8-shogaol, 10-shogaol and curcumin were the leading compounds in reducing major proinflammatory mediators and cytokines, and a simplified compound combination of 6-s, 10-s and curcumin showed the greatest potency in reducing LPS-induced NO production. Our study provides scientific evidence in support of the combined use of ginger and turmeric to alleviate inflammatory processes.     

Pharmacokinetics of [6]-gingerol after intravenous administration in rats

A high-performance liquid chromatographic method to determine [6]-gingerol, a pungent constituent of ginger, in rat plasma was developed and a pharmacokinetic study was performed in rats. Quantitative analysis with high reproducibility was achieved for [6]-gingerol over the concentration range of 0.2-40 micrograms/ml. After bolus intravenous administration at a dose of 3 mg/kg, the plasma concentration-time curve was described by a two-compartment open model. [6]-Gingerol was rapidly cleared from plasma with a terminal half-life of 7.23 min and a total body clearance of 16.8 ml/min/kg. Serum protein binding of [6]-gingerol was 92.4%.     

Pressurized liquid extraction of ginger (Zingiber officinale Roscoe) with bioethanol: an efficient and sustainable approach

To develop an efficient green extraction approach for recovery of bioactive compounds from natural plants, we examined the potential of pressurized liquid extraction (PLE) of ginger (Zingiber officinale Roscoe) with bioethanol/water as solvents. The advantages of PLE over other extraction approaches, in addition to reduced time/solvent cost, the extract of PLE showed a distinct constituent profile from that of Soxhlet extraction, with significantly improved recovery of diarylheptanoids, etc. Among the pure solvents tested for PLE, bioethanol yield the highest efficiency for recovering most constituents of gingerol-related compounds; while for a broad concentration spectrum of ethanol aqueous solutions, 70% ethanol gave the best performance in terms of yield of total extract, complete constituent profile and recovery of most gingerol-related components. PLE with 70% bioethanol operated at 1500 psi and 100 °C for 20 min (static extraction time: 5 min) is recommended as optimized extraction conditions, achieving 106.8%, 109.3% and 108.0% yield of [6]-, [8]- and [10]-gingerol relative to the yield of corresponding constituent obtained by 8h Soxhlet extraction (absolute ethanol as extraction solvent).     

Immunosuppressive activity of 8-gingerol on immune responses in mice

8-gingerol is one of the principal components of ginger, which is widely used in China and elsewhere as a food, spice and herb. It shows immunosuppressive activity on the immune responses to ovalbumin (OVA) in mice. In the present study, we found that 8-gingerol suppressed lipopolysaccharide (LPS) and concanavalin A (ConA)-stimulated splenocyte proliferation in vitro. In vivo, 8-gingerol not only signi?cantly suppressed Con A-, LPS- and OVA-induced splenocyte proliferation (P < 0.05) but also decreased the percentage of CD19+ B cells and CD3+ T cell (P < 0.05) at high doses (50, 100 mg/kg). Moreover, OVA-speci?c IgG, IgG1 and IgG2b levels in OVA-immunized mice were reduced by 8-gingerol at doses of 50, 100 mg/kg. These results suggest that 8-gingerol could suppress humoral and cellular immune responses in mice. The mechanism might be related to direct inhibition of sensitized T and B lymphocytes.     

Determination of Gingerols in Ginger by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

A rapid and sensitive ultra-high performance liquid chromatography–tandem mass spectrometry method was developed and validated for the determination of 6-, 8-, and 10-gingerol in the rhizomes of Zingiber officinale Rosc. The chromatographic separation was achieved isocratically on a C₁₈ (2.1 mm × 100 mm; 1.7 µm) column using acetonitrile:water (90:10 v/v) at a flow rate of 0.25 mL/min. Mass spectrometric transitions for 6-gingerol, 8-gingerol, and 10-gingerol occurred at m/z 277.2467/177.1470, 305.3196/177.1617, and 333.3614/177.1666, respectively. The analysis time was 3.0 min and the elution of 6-, 8-, and 10- gingerol occurred at 1.27, 1.51, and 1.80 min. The linear dynamic range was established over the concentration range of 2 and 1000 ng/mL for the analytes. The limits of detection and quantitation for 6-gingerol, 8-gingerol, and 10-gingerol were 0.921, 0.856, and 1.069 ng/mL, and 2.951, 2.727, and 3.013 ng/mL, respectively. The relative standard deviation and accuracy were found to be satisfactory. The results showed significant variation (0.176%–0.290% w/w) in gingerols among twelve accessions and that the Patna and Lucknow varieties contained the highest concentrations and were concluded to be superior. These results may be used to devise strategies for cultivating this plant for large scale production.     

Solvent extraction-spectrophotometric determination of trace amounts of ammonium, barium and potassium in a mixture by dicyclohexyl-18-crown-6 and orange II

A simple and sensitive extraction-spectrophotometric method for the determination of ammonium, barium and potassium in a mixture solution is reported. The DC18C6-ammonium-Orange II (DC18C6-NH(4)-OR II), DC18C6-Barium-Orange II (DC18C6-Ba-(OR II)(2)) and DC18C6-Potassium-Orange II (DC18C6-K-OR II) ternary complexes are quantitatively extracted into dichloromethane and their absorbances are measured at 483 nm. Linear calibration graphs were obtained over the concentration ranges of 0.05-3.00, 0.05-2.00 and 0.05-6.00 mug ml(-1) for ammonium, barium and potassium, respectively. The relative standard deviations for 1.0 mug ml(-1) of ammonium, barium and potassium are, respectively, 1.1, 1.6 and 2.50%. The limit of detection, LOD, for these three ions is reported to be 0.02, 0.01 and 0.02 mug ml(-1), respectively. The interfering effect of a large number of diverse ions on the determination of these three ions was studied. The method was applied to the Karon river raw water contaminated with industrial and house sewages. The results showed high potential of the recommended method for the determination of NH(4)(+), Ba(2+) and K(+) in water samples.