Solvent terminated natural deep eutectic solvent microextraction for concentration of curcuminoids in Curcumae Longae Rhizoma and turmeric tea.

A novel solvent terminated microextraction method based on a natural deep eutectic solvent (L-menthol and lactic acid at a molar ratio of 1:2) coupled with high-performance liquid chromatography was proposed, which was utilized for the separation and enrichment of bisdemethoxycurcumin, demethoxycurcumin and curcumin in Curcumae Longae Rhizoma and turmeric tea. The effects of independent parameters on extraction efficiency were optimized by single-factor analysis. Subsequently, four predominated parameters affecting the extraction procedure, including extractant volume, salt concentration, demulsifier consumption, and demulsification time, were further evaluated by a central composite design. Under the optimized conditions, the linear ranges of calibration curves were 0.005-0.5 µg/mL for bisdemethoxycurcumin, 0.004-0.4 µg/mL for demethoxycurcumin, and 0.0045-0.45 µg/mL for curcumin, respectively. In addition, the developed method provided low detection limits (0.1-0.4 ng/mL) and high enrichment factors (279-350). Its intra-day and inter-day precision were carried out by relative standard deviation ranging from 2.2 to 9.2%. Finally, the applicability of this method was assessed by the analysis of Curcumae Longae Rhizoma and turmeric tea samples. The results showed that these samples were detected successfully and the spiked recoveries over the range of 85.3-108.9% with relative standard deviations of 1.6-8.9% were attained, indicating its high relative recoveries with good precision in real sample analysis.

[Efficacy-related substances of blood-activating and stasis-resolving medicinals derived from Curcuma plants: a review].

Derived from Curcuma plants, Curcumae Longae Rhizoma, Curcumae Rhizoma, Wenyujin Rhizoma Concisum, and Curcumae Radix are common blood-activating and stasis-resolving medicinals in clinical practice, which are mainly used to treat amenorrhea, dysmenorrhea, chest impediment and heart pain, and rheumatic arthralgia caused by blood stasis block. According to modern research, the typical components in medicinals derived from Curcuma plants, like curcumin, demethoxycurcumin, bisdemethoxycurcumin, curdione, germacrone, curcumol, and ß-elemene, have the activities of hemorheology improvement, anti-platelet aggregation, anti-thrombosis, anti-inflammation, anti-tumor, and anti-fibrosis, thereby activating blood and resolving stasis. However, due to the difference in origin, medicinal part, processing, and other aspects, the efficacy and clinical application are different. The efficacy-related substances behind the difference have not yet been systematically studied. Thus, focusing on the efficacy-related substances, this study reviewed the background, efficacy and clinical application, efficacy-related substances, and "prediction-identification-verification" research method of blood-activating and stasis-resolving medicinals derived from Curcuma plants, which is expected to lay a theoretical basis for the future research on the "similarities and differences" of such medicinals based on integrated evidence chain and to guide the scientific and rational application of them in clinical practice.

[Breeding of a new cultivar "Chuanjianghuang 1" of Sichuan Dao-di herbs Curcumae Longae Rhizoma and Curcumae Radix].

The systematic breeding method was adopted to breed a new good cultivar of Curcuma longa, named "Chuanjianghuang 1". From 2014 to 2015, two consecutive years of multi-point test were carried out in Shuangliu, Chongzhou and Wenjiang. The biological characters, phenology, agronomic characters, yield and quality indexes of "Chuanjianghuang 1" were comprehensively evaluated. The results showed that compared with local traditional species, the rhizome yield of the new cultivar "Chuanjianghuang 1" increased by 20.61%.The average content of volatile oil was higher than 24.17% and the average content of curcumin in root tuber was higher than 26.62%. The yield of root tuber increased by 54.59%.The average content of volatile oil is higher than 36.28% and the average content of curcuminoids is higher than 25.31%. Compared with "Huangsi Yujin 1", "Chuanjianghuang 1" increased the average yield of rhizome by 123.68%,the volatile oil increased by an average of 7.69%and the curcumin content increased by an average of 58.23%. The average content of volatile oil is higher than 52.82% and the average content of curcuminoids in root tuber was higher than 38.34%. The new variety "Chuanjianghuang 1" has better yield than the local traditional species, and the internal quality of rhizome and root tuber is better. Compared with "Huangsi Yujin 1", the yield of rhizome is significantly increased, and the internal quality of rhizome and root tuber is better, especially the content of curcumin in rhizome and curcuminoids in root tuber is significantly higher than that of "Huangsi Yujin 1". "Chuanjianghuang 1" is high yield, good quality, good stability and strong adaptability, which is suitable for cultivation and promotion in Chengdu Jinma River Basin, such as Shuangliu, Chongzhou, Wenjiang.

[Nature-effect relationship research of Curcumae Longae Rhizoma,Curcumae Radix,and Curcumae Rhizoma based on nature combination].

Curcumae Longae Rhizoma,Curcumae Radix and Curcumae Rhizoma are different medicinal parts of the same plant.Nevertheless,they are different in medicinal effects due to the different Chinese herbal nature. In this study,traditional Chinese medicines database( TCMD2009),traditional Chinese medicine system( TCMSP),and Ch EMBL database were retrieved to screen the active components and targets,and construct the target PPI network. By a graph theoretic clustering algorithm identifying protein complex algorithm( IPCA),the protein modules were identified and analyzed by gene ontology( GO) enrichment. A comparative study of Curcumae Longae Rhizoma and Curcumae Radix illustrate that Curcumae Longae Rhizoma regulates blood coagulation through P2 RY12,GNG2 and other genes to exert the analgesic effect. Curcumae Radix regulates lipid metabolism,plasma lipoprotein particle levels,platelet activation,response to oxidative stress,apoptotic process through LDLR,APOB,PRKCA,SOD1,TP53 and other genes to perform a function in clearing the heart and cooling the blood. A comparative study of Curcumae Radix and Curcumae Rhizoma demonstrate that Curcumae Rhizoma on regulates the nervous system by GRIA2,GRIA4 and other genes to exert blood-breaking effect; Curcumae Radix regulates lipid metabolism,plasma lipoprotein particle levels,platelet activation,response to oxidative stress,apoptotic process by genes such as CALM1,LPL,APOB,SOD1 and TP53 to play the role of clearing heart and cooling blood. Cluster analysis of the protein interaction network of the nature combination is helpful to explain the intrinsic link between the nature combination and efficacy.

[High-throughput screening of ochratoxin A in Chinese herbal medicines using enzyme-linked immunoassay].

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.

[Research on standard decoction of processed Curcumae Longae Rhizoma containing volatile oil].

To prepare processed Curcumae Longae Rhizoma standard decoction, establish its quality standard, provides quality reference for formulated Jianghuang granules and lay a foundation for standard decoction research for the pieces containing volatile oil. Thirteen batches of processed Curcumae Longae Rhizoma standard decoctions were prepared according to the preparation requirements for standard decoction of Chinese herbal medicine containing volatile oil. With curcumin as the detection index, the transfer rate and extraction ratio were calculated; the pH value was determined and HPLC fingerprint analysis method was established. The results showed that the transfer rate ranged from 0.67% to 2.24%; the extraction rate was at a range of 9.95% to 18.36% and pH value was 3.86 to 6.05. Then the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012A) was used to analyze and compare the fingerprint. Sixteen common peaks were determined and three were identified including bisdemethoxycurcumin (peak 7), demethoxycurcumin (peak 8) and curcumin (peak 9). Moreover, the similarity was over 0.78 in the similarity evaluation of 13 batches of samples. HPLC fingerprint analysis method of processed Curcumae Longae Rhizoma standard decoction was established, showing good precision, stability and repeatability with certain identification effect, suitable for the quality control of formulated Jianghuang granules.

The extract of Curcumae Longae Rhizoma suppresses angiogenesis via VEGF-induced PI3K/Akt-eNOS-NO pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcumae Longae Rhizoma (CLR) is a safe natural herbal medicine, and which has been widely used for centuries as functional food and health products, but its effects on angiogenesis and related underlying mechanism remain unclear. AIM OF THE STUDY: The abnormal angiogenesis is closely related with various diseases, and therefore the precise control of angiogenesis is of great importance. The well-known angiogenic factor, vascular endothelial growth factor (VEGF), mediates angiogenesis and induces multiple signalling pathways via binding to VEGF receptor (VEGFR). The attenuation of VEGF-triggered angiogenic-related signalling pathways may relieve various diseases through suppression of angiogenesis. Here, we aimed to elucidate that CLR extract could exert striking anti-angiogenic activities both in vitro and in vivo. MATERIALS AND METHODS: The viability of human umbilical vascular endothelial cell (HUVEC) was examined by LDH and MTT assays. Migrative and invasive ability of the endothelial cells were independently evaluated by wound healing and transwell assays. The activities of CLR extract on in vitro angiogenesis was tested by tube formation assay. In vivo vascularization was determined by using zebrafish embryo model in the present of CLR extract. Western blotting was applied to determine the phosphorylated levels of VEGFR2, PI3K, AKT and eNOS. Besides, the levels of nitric oxide (NO) and reactive oxygen species (ROS) were separately evaluated by Griess assay and 2'7'-dichlorofluorescein diacetate reaction. In addition, the cell migrative ability of cancer cell was estimated by using cultured human colon carcinoma cells (HT-29 cell line), and immunofluorescence assay was applied to evaluate the effect of CLR extract on nuclear translocation of NF-κB p65 subunit in the VEGF-treated HT-29 cultures. RESULTS: CLR extract significantly suppressed a series of VEGF-mediated angiogenic responses, including endothelial cell proliferation, migration, invasion, and tube formation. Moreover, CLR extract reduced in vivo sub-intestinal vessel formation in zebrafish embryo model. Mechanistically, the extract of CLR attenuated the VEGF-triggered signalling, as demonstrated by decreased level of phosphorylated VEGFR2 and subsequently inactivated its downstream regulators, e.g. phospho-PI3K, phospho-AKT and phospho-eNOS. The production of NO and formation of ROS were markedly inhibited in HUVECs. Furthermore, CLR extract suppressed cell migration and NF-κB translocation in cultured HT-29 cells. CONCLUSIONS: These preclinical findings demonstrate that the extract of CLR remarkably attenuates angiogenesis and which has great potential as a natural drug candidate with excellent anti-angiogenic activity.

Evaluation of 60Co Irradiation on Volatile Components of Turmeric (Curcumae Longae Rhizoma) Volatile Oil with GC-IMS.

60Co irradiation is an efficient and rapid sterilization method. The aim of this work is to determine the changes in essential-oil composition under different irradiation intensities of 60Co and to select an appropriate irradiation dose with GC-IMS. Dosages of 0, 5, and 10 kGy of 60Co were used to analyze turmeric (Curcumae Longae Rhizoma) volatile oil after 60Co irradiation (named JH-1, JH-2, and JH-3). The odor fingerprints of volatile organic compounds in different turmeric volatile oil samples were constructed by headspace solid-phase microextraction and gas chromatography-ion mobility spectrometry (GC-IMS) after irradiation. The differences in odor fingerprints of volatile organic compounds (VOCs) were compared by principal component analysis (PCA). The results showed that 97 volatile components were detected in the volatile oil of Curcuma longa, and 64 components were identified by database retrieval. With the change in irradiation intensity, the volatile compounds in the three turmeric volatile oil samples were similar, but the peak intensity was significantly different, which was attributed to the change in compound composition and content caused by different irradiation doses. In addition, the principal component analysis showed that JH-2 and JH-3 were relatively correlated, while JH-1 and JH-3 were far from each other. In general, different doses of 60Co irradiation can affect the content of volatile substances in turmeric volatile oil. With the increase in irradiation dose, the peak area decreased, and so the irradiation dose of 5 kGy/min was better. It is shown that irradiation technology has good application prospects in the sterilization of foods with volatile components. However, we must pay attention to the changes in radiation dose and chemical composition.

Sources, morphology, phytochemistry, pharmacology of Curcumae Longae Rhizoma, Curcumae Radix, and Curcumae Rhizoma: a review of the literature.

Curcumae Longae Rhizoma (turmeric), Curcumae Radix and Curcumae Rhizoma are derived from the Curcuma species, and have gradually become three of the most commonly used medicinal herbs in China due to their different origins, processing methods and medicinal part. These three herbs have certain similarities in morphology, chemical composition, and pharmacological effects. All three of these herbs contain curcuminoids and volatile oil compounds, which exhibit anti-inflammatory, anti-tumor, antioxidant, and neuroprotective properties, although modern clinical applications have their own requirements. At present, there is no systematic guidelines for the clinical application of these three of Curcuma species; consequently, there is a high risk of unwanted phenomena associated with the mixing and indiscriminate use of these herbs. In this review, we focus predominantly on morphology, chemical composition, and the pharmacological activity of these three Curcuma herbs and summarize the current status of research in this field. Our goal is to provide a better understanding of clinical value of these Curcuma species so that we can provide reference guidelines for their further development, utilization and rational clinical application.

Feasibility of mixed herbal medicine for improving gastric function in an alcohol-induced gastritis model.

PURPOSE: Excessive exercise causes various gastric dysfunction. Gastritis is common among athletes who perform high-intensity training. Gastritis is a digestive disease involving mucosal damage caused by inflammatory reactions and oxidative stress. In this study, the effects of a complex natural extract on gastric mucosal damage and the expression of inflammatory factors were evaluated in an animal model of alcohol-induced gastritis. METHODS: A mixed herbal medicine (Ma-al-gan; MAG) was prepared with four natural products (Curcumae longae Rhizoma, Schisandrae chinensis Fructus, Artemisiae scopariae herba, and Gardeniae Fructus) identified by a systemic analysis using the Traditional Chinese Medicine Systems Pharmacology platform. The effects of MAG on alcohol-induced gastric damage were evaluated. RESULTS: MAG (10-100 µg/mL) significantly reduced the mRNA and protein levels of inducible nitric oxide synthase and cyclooxygenase-2 in lipopolysaccharide-stimulated RAW264.7 cells. MAG (500 mg/kg/d) effectively prevented alcohol-induced gastric mucosal injury in vivo. CONCLUSION: MAG regulates inflammatory signals and oxidative stress and is a potential herbal medicine for gastric disorders.

Curcumae longae Rhizoma (Jianghuang) extract reverses the 5-Fluoruracil resistance in colorectal cancer cells via TLR4/PI3K/Akt/mTOR pathway.

OBJECTIVES: Intensive efforts have been made in the area of identifying drug resistance modulators from traditional Chinese medicine. Various natural plant extracts have been reported for their reversal effect of drug resistance in cancers. This study was to assess the reversal potential of Curcumae longae Rhizoma extract (CLRE) in 5-Fluorouracil (5-Fu) resistance to colon cancer and explore its underly mechanism. METHODS: Increased concentrations of 5-Fu were used to culture SW480. A series of concentrations of CLRE were used to treat the 5-Fu resistant SW480 cells. WST-8 assay was used to detect the cell viability. Cell apoptosis was assessed by SuperView™ 488 Caspase-3 Assay Kit. The quantification of mentioned factors was archived by RT-qPCR. Network pharmacology analysis was used to explore the target of CLRE. RESULTS: 5-Fu resistant cell line (SW480/5-FuR) was established. The IC50 value of CLRE against SW480/5-FuR was 181.0 ± 14.12 µg/ml. CLRE can resensitize the SW480/5-FuR to 5-Fu by inhibiting cell growth. The combination treatment (CLRE+5-Fu) induced cell apoptosis via inhibition of bcl-2 and activation of caspase-3 and bax. Three active ingredients from CLRE were identified. TLR4 was targeted by these three ingredients and linked these ingredients to PI3K/Akt/mTOR pathway. The levels of TLR4, PI3K, AKT1, and mTORC1 mRNA were decreased when 5-Fu was combined with CLRE. CONCLUSIONS: CLRE could reverse 5-Fu resistance in colon cancer by inactivating TLR4/PI3K/AKT/mTORC1 pathway. This finding might provide a molecular basis and a valuable direction for further clinical applications and research for treating 5-Fu resistant colon cancer.

Curcumae Longae Rhizoma and Saussureae Radix Inhibit Nitric Oxide Production and Cannabinoid Receptor 2 Down-regulation.

BACKGROUND/AIM: The cannabinoid 2 (CB2) receptor is an important regulator of immunoinflammatory responses. Crude drugs commonly used in Japanese traditional Kampo medicine have displayed anti-inflammatory effects; however, few studies have reported that these effects are mediated via CB2 receptor signaling. Therefore, this study aimed to elucidate CB2 receptor-related anti-inflammatory regulation in crude drugs. MATERIALS AND METHODS: The ethanol extracts of 34 crude drugs listed in the Japanese Pharmacopeia were tested, and the inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production were evaluated in murine macrophage RAW 264 cells. RESULTS: The extracts of Curcumae Longae Rhizoma (dried rhizome of Curcuma longa) and Saussureae Radix (dried root of Saussurea lappa) significantly inhibited NO production and attenuated the LPS-induced decrease in CB2 receptor mRNA expression. CONCLUSION: Curcumae Longae Rhizoma and Saussureae Radix can modulate the CB2-receptor-related anti-inflammatory regulation in macrophages.

Pharmacokinetic based study on "lagged stimulation" of Curcumae Longae Rhizoma - Piper nigrum couplet in their main active components' metabolism using UPLC-MS-MS.

BACKGROUND: Curcumae Longae Rhizoma is one of the commonly used traditional Chinese medicines, which has multiple biological activities such as relieving stagnation and stasis, pain alleviation, curing amenorrhea and wounds. However, its main active component-curcumin has poor absorption and very fast metabolism in body. To solve this problem, Piper nigrum was introduced for its ability to strengthen bioavailability of other compounds. PURPOSE: In most cases of TCM couplets, all ingredients were prepared and taken simultaneously, which in our opinion did not take full advantage of their interactions. Therefore, order of administration should be adjusted according to pharmacokinetic parameters of the ingredients, which the ones act as supplement can first be taken, and main therapeutic components followed when the former reached its peak. METHOD: the extract of Piper nigrum (containing at least 95% piperine) was taken by rats 6h before taking Curcumae Longae Rhizoma extract (containing at least 95% curcumin). Then, a UPLC-MS-MS method was developed to determine their content in plasma simultaneously. Determination was carried out by on a C18 column within 5min by isocratic elution using 0.2% formic acid and acetonitrile (50:50, v/v). Tandem mass detection was conducted by selective reaction monitoring (SRM) via electrospray ionization (ESI) source in positive mode. Samples were pre-treated by liquid-liquid extraction (LLE), and verapamil was used as internal standard (IS). RESULTS: For both curcumin and piperine, the proposed method had good linearity (r2=0.999) within the concentration range of 1-1000ng/ml, with good recovery, precision and stability. The lower limit of quantification (LLOQ) was 1ng/ml. As pharmacokinetic data indicated, Maximum concentration (Cmax) of curcumin increased significantly to 394.06; the time reach maximum concentration (Tmax) and elimination half-life (T1/2) were 0.5 and 0.67h, respectively; CONCLUSION: The results provide a good strategy for the investigation of TCM formula especially the couplets, as well as a fast, selective and sensitive UPLC-MS-MS method determining active components in-vivo. Furthermore, the finding of "lagged stimulation" suggested that the use of complex formula should take pharmacokinetics into much more careful consideration.