Semen Cuscutae Research Articles

Semen Cuscutae Administration Improves Hepatic Lipid Metabolism and Adiposity in High Fat Diet-Induced Obese Mice.

Since arginase has been shown to compete with nitric oxide (NO) synthase, emerging evidence has reported that arginase inhibition improves obesity by increasing NO production. Semen cuscutae (SC), which is a well-known Chinese medicine, has multiple biological functions such as anti-oxidant function and immune regulation. In this study, we investigated whether the SC as a natural arginase inhibitor influences hepatic lipid abnormalities and whole-body adiposity in high-fat diet (HFD)-induced obese mice. The lipid accumulation was significantly reduced by SC treatment in oleic acid-induced hepatic steatosis in vitro. Additionally, SC supplementation substantially lowered HFD-induced increases in arginase activity and weights of liver and visceral fat tissue, while increasing hepatic NO. Furthermore, elevated mRNA expressions of sterol regulatory element-binding transcription factor 1 (SREBP-1c), fatty-acid synthase (FAS), peroxisome proliferator-activated receptor-gamma (PPAR-γ)1, and PPAR-γ2 in HFD-fed mice were significantly attenuated by SC supplementation. Taken together, SC, as a novel natural arginase inhibitor, showed anti-obesity properties by modulating hepatic arginase and NO production and metabolic pathways related to hepatic triglyceride (TG) metabolism.

Development of a HPLC-MS/MS Method to Determine the 13 Elements of Semen Cuscutae and Application to a Pharmacokinetic Study in Rats

This study developed a method for simultaneous determination of 13 elements of Semen Cuscutae (quercitrin, quercetin, hyperoside, caffeic acid, chlorogenic acid, luteolin, apigenin, kaempferol, isoquercitrin, cryptochlorogenic acid, isorhamnetin-3-O-glucoside, astragalin, and rutin) in rat plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in the negative MRM mode. The analytes were analyzed with CORTECS®C18 column (4.6 × 150 mm, 2.7 μm) with mobile phases consisting of 0.1% formic acid in water (A) and acetonitrile (B). The intra- and interday precision of the target compounds were expressed as relative standard deviation (RSD) in the range of 0.5%–10.4%, and the accuracy of the target compounds was expressed as relative error (RE) not exceeding ±14.5% for all analytes. In the meantime, the extraction recovery of the target compounds in plasma samples ranged from 87.4% to 106.2% and matrix effect from 81.0% to 115.5%. The established method was successfully accomplished for the pharmacokinetic study of the analytes in rat plasma samples following oral administration of Semen Cuscutae extract, and the pharmacokinetic parameters of seven compounds were obtained.

Rapid and undamaged identification of the Semen cuscutae and its adulterants based on image analysis and electronic nose analysis

In this paper, image analysis and electronic nose analysis were used to develop a rapid, reliable and undamaged method of identifying the Semen cuscutae and its adulterants including radish seed and Sinapis alba seeds. The results showed that the highest identification rate was 100% for the training set and 96.5% for the test set based on image analysis and various chemometric techniques including principal component analysis, linear discriminant analysis (LDA), k-nearest neighbor, random forests, artificial neural network and support vector machine analysis. LDA analysis based on electronic nose analysis exhibited better discrimination result, ranging from 95.5–100% for the correct classification rate and 95.4–100% for the cross-validation rate, respectively. LDA model based on electronic nose data from 16 to 30 s was the best, and both the correct classification rate and cross-validation rate reached 100%. These results provided a simple, fast and non-destructive method to identify the true and false of Semen cuscutae, which can serve as a reference to identify the authenticity of the medicinal plants.

Study on mechanism of Cuscutae Semen flavonoids in improving reproductive damage of Tripterygium Glycosides Tablets in rats based on high-throughput transcriptome sequencing

Tripterygium Glycosides Tablets has good anti-inflammatory and immunomodulatory activities,but its reproductive damage is significant. Previous studies of the research group have found that Cuscutae Semen flavonoids can improve spermatogenic cell damage caused by Tripterygium Glycosides Tablets by regulating spermatogenic cell cycle,apoptosis and related protein expression,but the mechanism of action at the gene level is still unclear. In this study,Illumina high-throughput sequencing platform was applied in transcriptional sequencing of spermatogenic cells of rats after the intervention of Cuscutae Semen flavonoids and Tripterygium Glycosides Tablets. Differentially expressed genes were screened out and the GO enrichment and KEGG pathway analysis of differentially expressed genes were conducted to explore the mechanism of Cuscutae Semen flavonoids in improving reproductive injury caused by Tripterygium Glycosides Tablets. The results showed that 794 up-regulated genes and 491 down-regulated genes were screened in Tripterygium Glycosides Tablets group compared with the blank group. Compared with Tripterygium Glycosides Tablets,440 up-regulated genes and 784 down-regulated genes were screened in the Cuscutae Semen flavonoids+Tripterygium Glycosides Tablets group. Among them,the gene closely related to reproductive function is DNMT3 L. Analysis of GO function and KEGG signaling pathway enrichment showed that the above differentially expressed genes were mainly enriched in cell,cell process,catalytic activity,binding,ovarian steroid synthesis,thyroid hormone and other functions and pathways. The thyroid hormone signaling pathway was the common enrichment pathway of the two control groups. In a word,Cuscutae Semen flavonoids has a good treatment effect on male reproductive damage caused by Tripterygium Glycosides Tablets. The mechanism may be closely related to up-regulation of DNMT3 L genes and intervention of thyroid hormone signaling pathway. At the same time,the discovery of many different genes provides valuable information for study on the mechanism of Cuscutae Semen flavonoids and Tripterygium Glycosides Tablets compatibility decreasing toxicity and increasing efficiency.

A comprehensive analysis of 201 pesticides for different herbal species-ready application using gas chromatography–tandem mass spectrometry coupled with QuEChERs

Developing an analysis of multi-pesticide residues for different herbal species-ready applications is a challenge. In the present work, a comprehensive analysis was proposed for rapid detection of 201 pesticides in various medicinal herbs. Samples were extracted and cleaned up with a high throughput pretreatment approach (modified QuEChERS), and then detected by gas chromatograph coupled to an electron impact ionization triple quadrupole mass spectrometer (GC-EI-MS/MS). The clean-up procedure has been optimized using four types of representative medicinal herbs with different primary or secondary metabolites. Moreover, a mixture of analyte protectants (APs) was to improve the peak shape and intensity of some compounds. The performance of the method was validated according to the European Union SANTE/11813/2017 regulatory guidelines. The limit of quantification (LOQ) was determined to be ≤10 ng mL−1, and the recovery was between 70.0%–120.0%, with ≤20% RSD for the majority of pesticides. Sixty samples belonging to different species of medicinal herbs (such radix, flos, cortex, fructus, and seeds) were analyzed to evaluate the applicability of the optimized method. High frequency of chlorpyrifos was found in Citri reticulatae pericarpium, Crataegi fructus and Cuscutae semen samples.

An Established HPLC-MS/MS Method for Evaluation of the Influence of Salt Processing on Pharmacokinetics of Six Compounds in Cuscutae Semen.

A sensitive and effective method was developed for clarifying the pharmacokinetic properties of six compounds (including hyperin, chlorogenic acid, neochlorogenic acid, p-coumaric acid, astragalin, and isoquercitrin) in two processed Cuscutae Semen samples by high performance liquid chromatography mass spectrometry (HPLC-MS/MS). The six compounds were separated by acetonitrile and 0.1% formic acid-water on an Agilent Eclipse plus C18 column (4.6 mm × 100 mm, 1.8 μm). All compounds were analyzed with negative ion mode in multiple reaction monitoring (MRM). The lower limits of quantification (LLOQ) of hyperin, astragalin, neochlorogenic acid, chlorogenic acid, isoquercitrin, and p-coumaric acid were 1, 0.1, 4, 0.1, 2, and 4 ng·mL−1, respectively. The validated approach was effectively used for the pharmacokinetics of six compounds of two processed Cuscutae Semen samples after oral administration to rat. The results indicated that salt processing could improve the adsorption and bioavailability of astragalin in Cuscutae Semen.

Using single-patient (n-of-1) trials to determine effectiveness of traditional Chinese medicine on chemotherapy-induced leukopenia in gastric cancer: a feasibility study.

Gastric cancer has been the second cause of cancer death worldwide. Chemical comprehensive treatment programs primarily were the main therapy method with modest efficacy to gastric cancer. Traditional Chinese medicine (TCM) has been reported to alleviate adverse events induced by chemotherapy, but has not yet developed clinical trials to test and needs scientific evidence for making policy. Single-patient (N-of-1) trials might be an eligible study design for TCM since it well represented the individualized treatment philosophy of TCM. The aim of this study is to obtain information necessary to design a more series trial. Individuals who underwent gastrectomy were included. Each patient suffered 3-week standard chemotherapy and 3-day treatment periods (decoction with Astragalus mongholicus and Semen Cuscutae or placebo: decoction without Astragalus mongholicus and Semen Cuscutae). Each trial lasted up to a maximum of 30 weeks or a minimum of 20 weeks. Staffs and participants were blinded to the randomization. This study was approved by Ethics Committee of First Hospital, Lanzhou University in November, 2014. From August, 2014 to March, 2015, 6 participants were included. There were 16 cycles compared between intervention and control decoction (2.28, 95% CI: 1.24-5.47), P<0.0001. The quality of life (QoL) score after the trial was reported is a little higher than before, t=3.87, P=0.01. Two participants reported symptoms had improved after taken trial decoction. This is the first N-of-1 trials of testing the effectiveness of TCM decoction on alleviative treatment to gastric cancer. The feasibility study will help to develop a practical design for the more series trial.

Development and Validation of HPLC-PDA Method and Pattern Recognition Analyses Using Eight Marker Compounds for the Quality Control Between the Seeds of Cuscuta chinensis Lam. and Cuscuta japonica Choisy

Cuscuta chinensis Lam. and Cuscuta japonica Choisy are parasitic plants. C. chinensis seeds were traditionally used for treatment of kidney and liver deficiencies. C. japonica seeds were used as tonic medicine to improve liver function and strengthen kidneys, treatment of high blood pressure, chronic diarrhea, and sore eyes. Cuscutae Semen are seeds of only C. chinensis in Korean Herbal Pharmacopoeia (K.H.P.). The developed HPLC-PDA method easily, accurately, and sensitively quantified using eight marker compounds [hyperoside (1), astragalin, (2), quercetin (3), kaempferol (4), chlorogenic acid (5), 3,4-di-O-caffeoylquinic acid (6), 1,5-di-Ocaffeoylquinic acid (7), and 4,5-di-O-caffeoylquinic acid (8)]. In addition, the method may be used to distinguish seeds between C. chinensis Lam. and C. japonica Choisy. Furthermore, the result from the current study was applied to clarify samples between steam processed and unprocessed samples of C. chinensis by pattern analysis.

Data mining analysis of the ointment medication by Prof. Yang Zhimin treating yang deficiency and fatigue state patients

Objective The purpose of this research is to use modern statistical methods, to analyze the rules of cointment by Prof. Yang Zhimin treating yang deficiency and fatigue state patients, to increase the pertinence and to improve the clinical efficacy. Methods This paper analyzed 62 cases of Prof. Yang Zhimin in the treatment of the patients with yang deficiency and fatigue state from October 2010 to January 2013. The analyses were frequency analysis and association rules. Results Among the patients with yang-deficiency and fatigue, 193 Chinese medicine were used. The was the highest frequency. Dampness-transforming medicine accounts for about 22.2% in the top 36 frequency of medicine. Based on analyzing association rules, we found that the compatibility of medicine which were used more frequently came from Sini decoction, Danggui Sini decoction and Jinkui Shenqi pills, such as monkshood and dried monkshood and roasted liquorice,dried ginger and roasted monkshood and monkshood and cornel, monkshood and prepared rehmannia root and monkshood and yam. Futhermore, we found two new core prescription. One was Qian Yang Dan combined with Sini decoction consist of monkshood, roasted liquorice, dried ginger, fructus amomi.The other one was Tu Si Jian consist of yam, semen cuscutae, angelica, roasted liquorice. Conclusions Invigorating spleen and kidney, warming yang and dissipating dampness and hiding yang and Seeking Yang in Yin are the habits and rules of the ointment medication by Prof. Yang Zhimin treating yang deficiency and fatigue state patients. Key words: Unguentum (TCD); Yang Zhimin; Yang deficiency; Fatigue; Data mining

Total flavonoids from Semen Cuscutae target MMP9 and promote invasion of EVT cells via Notch/AKT/MAPK signaling pathways

Miscarriage is a common condition during pregnancy and its mechanisms remain largely unknown. Extravillous trophoblast (EVT) cell invasion is required to maintain normal pregnancy and its malfunction has been proposed as a major cause for miscarriage. Homeostasis of matrix metalloproteinase 9 (MMP9) is a key to regulate EVT cell invasion. Total flavonoids from Semen Cuscutae (TFSC) have been applied clinically used for preventing or treating miscarriage in the past. Given its potential clinical benefit on preventing miscarriage, this study aims at examining the therapeutic effect of TFSC in the prevention of premature birth by upregulating MMP9 and promote EVT cell invasion. HTR-8 cells migration and invasion functions were analyzed using wound healing and transwell assays. The regulatory effect of TFSC on MMP9 expression and relevant signaling pathways were analyzed by Western Blot. The results show compared to control group, TFSC significantly promoted the migration of EVT cells in a dose and time-dependent manner. The migration and invasion of EVT cells were maximized at the highest dosage of 5 μg/ml of TFSC. The expression of MMP9 in EVT cells was significantly increased after TFSC treatment. Furthermore, cells treated with TFSC significantly upregulated protein expressions in Notch, AKT and p38/MAPK signaling pathways. We believe TFSC can promote the migration and invasion of EVT cells by increasing MMP9 expression, and prevent miscarriage by activating Notch, AKT, and MAPK signaling pathways.

Cuscuta chinensis and C. campestris Attenuate Scopolamine-Induced Memory Deficit and Oxidative Damage in Mice

The seeds of Cuscuta chinensis Lam. and C. campestris Yuncker have been commonly used as Chinese medical material for preventing aging. Our previous studies have found that C. chinensis and C. campestris possess anti-inflammatory activities in rodents. However, their other biological activities, such as memory-improving properties, have not yet been explored. In the present study, we examined the memory-improving effects of the extracts of C. chinensis and C. campestris on scopolamine (SCOP)-induced memory deficit and explored their underlying mechanism in mice. Both Cuscuta species improved SCOP-induced memory deficits in the passive avoidance test, elevated plus-maze, and spatial performance test of the Morris water maze in mice. In addition, compared with mice injected with SCOP, mice pretreated with both Cuscuta species stayed for a longer time on the platform for the probe test of the Morris water maze. Moreover, both Cuscuta species reduced brain acetylcholinesterase activity and malondialdehyde levels that were increased by SCOP, and the species restored the activities of antioxidant enzymes (superoxide dismutase and catalase) and the levels of glutathione that were decreased by SCOP in the brains of mice. Both Cuscuta species further decreased brain interleukin-1β and tumor necrosis factor-α levels that were elevated by SCOP. We demonstrated that both Cuscuta species exhibited a protective activity against SCOP-induced memory deficit, cholinergic dysfunction, oxidative damage, and neuroinflammation in mice, and C. campestris has better potential than C. chinensis. In addition, we provided evidence that the seeds of C. campestris can be used as Cuscutae Semen in Traditional Chinese Medicine.

Dienzyme-assisted salting-out extraction of flavonoids from the seeds of Cuscuta chinensis Lam.

The extraction process involving enzymatic treatment and salting-out extraction was developed to accelerate dissolution of flavonoids from the hard compact seeds of Cuscuta chinensis Lam. A combination of enzymes (cellulase and protease) provided higher flavonoid yields than single enzyme. The optimum enzymatic conditions were 5% acidic cellulase, 5% neutral protease, pH 4.5, 45 °C, and 45 min. The salting-out extraction used ammonium sulfate and ethanol (1:2, w/v). Flavonoid yields as high as 18.75 mg/g were obtained from Semen cuscutae under the optimum conditions. Results also showed that the antioxidant properties of the flavonoids were correlated with their contents dirrectly guided by the scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. The extraction process could be used for mass production of flavonoids.

High resolution UPLC-MS/MS method for simultaneous separation and determination of six flavonoids from semen cuscutae extract in rat plasma: application to comparative pharmacokinetic studies in normal and kidney-deficient rats

Semen Cuscutae, which mainly consisted of flavonoids, is a traditional Chinese herbal medicine and used for nourishing the liver and kidneys. The aim of this study was to develop a sensitive and selective UPLC-MS/MS method for simultaneous separation and determination of six main active renoprotective components of Hyperoside, Astragalin, Isoquercitrin, Quercitrin, Quercetin, and Kaempferol from Semen Cuscutae in rat plasma, and to reveal the pharmacokinetic differences between normal and kidney deficient rats. The validated method has been successfully applied to comparing pharmacokinetic profiles of the six analytes in rat plasma. The results indicated that there was significant difference in pharmacokinetic parameters of the six analytes between two groups, while absorptions in kidney deficient group were significantly lower than those in normal group. This study would be helpful for evaluating the Semen Cuscutae as renoprotective drug candidates for pre-clinical and clinical research.

Compounds Identification in Semen Cuscutae by Ultra-High-Performance Liquid Chromatography (UPLCs) Coupled to Electrospray Ionization Mass Spectrometry.

Semen Cuscutae is commonly used in traditional Chinese medicine and contains a series of compounds such as flavonoids, chlorogenic acids and lignans. In this study, we identified different kinds of compositions by ultra-high-performance liquid chromatography (UPLC) coupled to electrospray ionization mass spectrometry (MS). A total of 45 compounds were observed, including 20 chlorogenic acids, 23 flavonoids and 2 lignans. 23 of them are reported for the first time including 6-O-caffeoyl-β-glucose, 3-O-(4′-O-Caffeoylglucosyl) quinic acid, etc. Their structures were established by retention behavior, extensive analyses of their MS spectra and further determined by comparison of their MS data with those reported in the literature. As chlorogenic acids and flavonoids are phenolic compounds that are predominant in Semen Cuscutae, in conclusion, phenolic compounds are the major constituents of Semen Cuscutae.

Bushen Huoxue Recipe Alleviates Implantation Loss in Mice by Enhancing Estrogen-Progesterone Signals and Promoting Decidual Angiogenesis Through FGF2 During Early Pregnancy.

Bushen Huoxue recipe (BSHXR) is a classic Chinese herbal prescription for nourishing the kidney and activating blood circulation. It consists of six herbs: Astragali radix, Angelicae sinensis radix, Ligustici Chuanxiong Rhizoma, Cuscutae semen, Taxilli Herba, and Dipsaci Radix, and the main active constituents of BSHXR are ferulic acid, calycosin-7-glucopyranoside, hyperoside, quercitrin, and asperosaponin VI. In clinical practice, BSHXR is traditionally used to treat failed pregnancy and its complications. However, little is known about the underlying mechanism of BSHXR for the treatment of implantation loss during early pregnancy. In the current study, controlled ovarian hyperstimulation was induced in mice as our implantation loss model, and we evaluated the effects of BSHXR on implantation, decidualization, decidual angiogenesis, and reproductive outcome. We showed that BSHXR could regulate the supraphysiological levels of serum estrogen and progesterone observed in these mice, and also act on estrogen and progesterone receptors in the stroma and epithelium. BSHXR also enhanced FGF2 expression in the vascular sinus folding area of the decidua, thus potentially reducing implantation loss during early pregnancy and contributing to placentation and survival of the fetuses. Taken together, our findings provide scientific evidence for the application of BSHXR in the clinic as a treatment for implantation loss during early pregnancy, and warrant further investigation of BSHXR as an effective treatment for failed pregnancy and its complications.

Differences in the Mechanisms by Which Yang-Invigorating and Qi-Invigorating Chinese Tonifying Herbs Stimulate Mitochondrial ATP Generation Capacity

According to Chinese medicine theory, Yang/Qi plays a pivotal role in driving physiological functions in the body, these being highly dependent on mitochondrial ATP production. Consistent with this, Yang/Qi-invigorating Chinese tonifying herbs have been found to stimulate mitochondrial ATP generation capacity (ATP-GC) in H9c2 cardiomyocytes. In the present study, we have demonstrated that Yang-invigorating Chinese tonifying herbs (namely, Eucommiae Cortex, Cibotii Rhizoma, Dipsaci Radix, Cynomorii Herba, Cistanches Herba, Cuscutae Semen, EpimediiHerba and Morindae Radix) and Qi-invigorating Chinese tonifying herbs (namely, Ginseng Radix, Pseudostellariae Radix, Quinquefolii Radix, Codonopsis Radix, Astragali Radix, Atractylodis Rhizoma, Juiubae Fructus, Fici Simplicissimae Radix and Dioscoreae Oppositae Radix) act by different mechanisms to stimulate mitochondrial ATP-GC. While Yang-invigorating herbs fluidize mitochondrial membranes and thus stimulate ATP-GC, Qi-invigorating herbs can enhance cellular glutathione status and increase ATP-GC. The different mechanisms by which Yang-invigorating herbs and Qi-invigorating herbs stimulate mitochondrial ATP-GC may serve as the basis for establishing biomarkers for Yang/Qi-invigorating herbs and herbal health products in general.

Features of Professor Ma Kun's medication in treating ovulatory infertility

In order to analyze Professor Ma Kun's medication in treating anovulatory infertility, her prescriptions for treating anovulatory infertility in 2012-2015 were collected. The medication features and the regularity of prescriptions were mined by using traditional Chinese medicine inheritance support system, association rules, complex system entropy clustering and other mining methods. Finally, a total of 684 prescriptions and 300 kinds of herbs were screened out, with a total frequency of 11 156 times; And 68 core combinations and 8 new prescriptions were mined. The top three frequently used herbs by effect were respectively tonic herb, blood circulation promoting herb, and Qi-circulation promoting herb. The top three tastes were sweetness, bitterness and pungent flavor. The results showed 28 herbs with a high frequency of ≥100.The top 10 frequently used herbs were respectively Angelica Sinensis Radix, Cyperi Rhizoma, Chuanxiong Rhizome, Paeoniae Radix Rubra, Cyathulae Radix, Taxilli Herba, Cuscutae Semen, Codonopsis Radix, Ligustri Lucidi Fructus, Paeoniae Albaand Paeoniae Radix Alba. The association rules analysis showed commonly used herbal pairs, including Rehmanniae Radix Preparata-Chuanxiong Rhizome, Rehmanniae Radix Preparata-Angelica Sinensis Radix, Cuscutae Semen-Dipsaci Radix. In conclusion, Professor Ma has treated anovulatory infertility by nourishing the kidney and activating blood throughout the treatment course, and attached the importance to the relationship between Qi and blood and there gulation of liver, spleen and kidney in treating anovulatory infertility.

Varieties textual research, resource investigation and identification of commercial drugs on Cuscutae Semen

Through the textual research, resource investigation, literature reviews (including Flora of China, municipal Flora, pharmacopoeia of China and municipal drug standards) and identification of commercial drugs on Cuscutae Semen, it was found the species described in the herbal textual was Cuscuta chinensis, with good quality from Shandong and Henan Province. The identification of commodities showed the majority drugs were from C. australis, varied from the ancient herbal textuals .Mordern literature reviews indicate that it was necessary to strengthen the research on Cuscutae Semen from C. australis, C. chinensis and C. japonica because of their differences in resources, macroscopical and microscopical characters, while wrong descriptions in some literatures. It was suggested that the two species (C. australis and C. chinensis) should be separated in pharmacopoeia of China. The study provides scientific basis for the development and utilization of Cuscutae Semen.

Purification, characterization and biological effect of reversing the kidney-yang deficiency of polysaccharides from semen cuscutae

Semen cuscutae is a well-known Chinese medicine which has been used to nourish kidney. It is the first study to demonstrate that the polysaccharides from semen cuscutae showed significant activity of nourishing kidney-yang by increasing the levels of testosterone and estradiol, decreasing the level of blood urea nitrogen, improving immune function, possessing antioxidant effect. Three homogeneous polysaccharides were obtained by DEAE-cellulose and Sephacryl S-400 which were named as C-7WR1, C-7WR2 and C-7WR3 with average molecular weight of 7.59×104, 3.23×104 and 2.25×104 respectively. C-7WR1 was composed of fructose: mannose=0.02:1. C-7WR2 was composed of fructose: mannose: xylose: arabinose=0.01:1:0.14:0.33. C-7WR3 was composed of fructose: mannose: xylose: arabinose=0.01:1:0.10:0.47. They mainly contained mannose. Their fourier transform infrared features were similar. They all had no nucleic acid and protein.

Authenticity Survey of Cuscutae Semen on Markets Using DNA Barcoding

ObjectiveUsing authentic raw herbal materials is fundamental to herbal medicine quality. Cuscuta chinensis and C. australis are two important species of Cuscutae Semen recorded in Chinese Pharmacopoeia. Due to having tiny bodies of seeds, it is extremely difficult to differentiate them from adulterants and closely related species by morphologic characteristics, leading to serious safety problems. MethodsIn this study, we developed a fast and efficient method to identify Cuscutae Semen on the market. First, a total of 207 ITS2 sequences representing 45 related species of Cuscutae Semen were collected to construct a standard DNA barcode database, then 33 commercial samples purchased from markets were analyzed by BLAST, and Neighbor-joining tree was used to verify the efficacy of the database. ResultsThe percentage of counterfeits and adulterants in the 33 commercial samples were up to 69.7%, and only 10 commercial products were found to be genuine. The adulterated species included 11 species (Amaranthus hybridus, Brassica carinata, Brassica juncea var. megarrhiza, Chenopodium album, Corispermum heptapotamicum, Cuscuta alata, Cuscuta japonica, Cuscuta monogyna, Foeniculum vulgare, Glycine max, and Medicago sativa). ConclusionDNA barcoding is a fast and efficient method to identify Cuscutae Semen on the market.