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ObjectiveTo compare the effects of different drying methods on volatile components of Pseudostellariae Radix. MethodThe samples were dried by different methods， including air drying， sun drying， hot air drying （40， 60， 80 ℃） and vacuum freeze drying. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to compare the changes of volatile components in the samples after different treatments. The samples were incubated at 80 ℃ and 500 r·min-1 for 15 min， the injection temperature was 85 ℃， the injection volume was 200 μL， the flow rate of carrier gas was from 2 mL to 150 mL during 20 min， and the temperature of IMS detector was 60 ℃. SE-54 capillary column （0.32 mm×30 m， 0.25 μm） was used， the column temperature was 60 ℃， and the analysis time was 35 min. The differential spectra of volatile components were constructed and analyzed by principal component analysis （PCA）. ResultA total of 37 volatile components were identified from dried Pseudostellariae Radix. The number of compounds in descending order was ketones， aldehydes and alcohols. There were some differences in the volatile components in samples dried by different methods. And the volatile components in samples with sun drying， air drying and hot air drying at 40 ℃ were similar， compared with other drying methods， vacuum freeze drying and hot air drying at 80 ℃ had great effects on the volatile components of Pseudostellariae Radix， and the compounds in the samples with vacuum freeze drying were the least. ConclusionIn this study， GC-IMS for the detection and analysis of volatile components in Pseudostellariae Radix is established， which has the characteristics of high efficiency， nondestructive inspection and simple sample processing. This method can be used for the distinction of Pseudostellariae Radix dried by different methods. And hot air drying at 40 ℃ can effectively retain the volatile components of Pseudostellariae Radix， and achieve similar flavor to samples with sun drying and air drying.
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This study aimed to establish a method for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigate its contamination with mycotoxins, so as to provide technical guidance for monitoring the quality of Chinese medicinal materials and medication safety. The sample was extracted with 80% acetonitrile in an oscillator for 1 h, purified using the modified QuEChERS purifying agent(0.1 g PSA + 0.3 g C_(18) + 0.3 g MgSO_4), and separated on a Waters HSS T3 chromatographic column(2.1 mm×100 mm, 1.8 μm). The gradient elution was carried out with 0.1% formic acid in water and acetonitrile, followed by the scanning in the multi-reaction monitoring(MRM) mode and the analysis of mycotoxin contamination in 26 Pseudostellariae Radix samples. The recovery rates of the established method were within the range of 82.17%-113.6%, with the RSD values less than 7% and the limits of quantification(LOQ) being 0.019-0.976 μg·kg~(-1). The detection rate of 14 mycotoxins in 26 batches of medicinal materials was 53.85%. The detection rate of sterigmatocystin(ST) was the highest, followed by those of zearalenone(ZEN), aflatoxin G_2(AFG_2), fumonisin B_1(FB_1), HT-2 toxin, and nivalenol(NIV). Their respective detection rates were 38.46%, 26.92%, 23.08%, 11.54%, 11.54%, and 7.69%, with the pollution ranges being 1.48-69.65, 0.11-31.05, 0.11-0.66, 0.28-0.83, 20.86-42.56, and 0.46-1.84 μg·kg~(-1), respectively. The established method for the detection of 14 mycotoxins is accurate, fast and reliable. The research results have very important practical significance for guiding the monitoring and prevention and control of exogenous fungal contamination of Chinese medicinal materials.
Subject(s)
Aflatoxins/analysis , Chromatography, High Pressure Liquid/methods , Drug Contamination , Food Contamination/analysis , Mycotoxins/analysis , Plant Roots/chemistry , Tandem Mass Spectrometry/methodsABSTRACT
Trading volume in Chinese medicinal materials market has gradually declined, and origin market trade has become a trend. Studying the degree of market integration is helpful to understand the operation of origin market of Chinese medicinal materials, which is of great significance for rational planning of the production of Chinese medicinal materials and improving market efficiency. In this study, four varieties (Lonicerae Japonicae Flos, Lycii Fructus, Isatidis Radix and Pseudostellariae Radix) were selected to study the market integration degree of Chinese medicinal materials. Based on the price index data from 2013 to 2019, this paper makes an empirical analysis by using co-integration test, error correction model and Granger causality test. The results showed that the long-term integration of Chinese medicinal materials market was relatively high, but the short-term integration was far lower than the long-term integration. Finally, the author puts forward four policy suggestions: improving the price information platform conduction of Chinese medicinal materials, building high-quality brand of Chinese medicinal materials in non-authentic producing areas, speeding up the establishment quality traceability system and building modern logistics system of Chinese medicinal materials.
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OBJECTIVE: To investigate the differences in the accumulation and biosynthesis of metabolites in different lactic acid of Pseudostellariae Radix. METHODS: 1H-NMR based metabolomic approach combined with multivariate statistical analysis was used to investigate the chemical compositions and find the different metabolites in cultivated Pseudostellariae Radix and its wild-type. RESULTS: Thirty-five metabolites were identified in the 1H-NMR spectra, and 15 of which showed remarkable differences. The multivariate statistical analysis showed that the cultivated Pseudostellariae Radix and its wild-type could be distinguished obviously. The contents of isoleucine, linolenic acid, lactic acid, alanine, lysine, glutamic acid, glutamine, acetoacetic acid, succinic acid, γ-aminobutyric acid, phenylalanine, and sucrose in cultivated Pseudostellaria heterophylla were lower than its wild-type, while the cultivated Pseudostellariae Radix contained more xylose, raffinose, and fumaric acid. CONCLUSION: This study will provide basic information for exploring the quality formation mechanism and revealing the accumulation and biosynthesis of metabolites in different ecotypes of Pseudostellariae Radix.
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Objective: To clone the gibberellin 2-oxidase (GA2ox) genes from Pseudostellaria heterophylla and perform the bioinformatic and expression mode analysis. Methods: According to P. heterophylla transcriptome annotation, two transcript codings of GA2ox were identified. The full-length cDNA PhGA2ox1 and PhGA2ox8 were determined using RT-PCR. Then the bioinformatic analysis of these genes and encoded proteins were performed. The coding sequences of PhGA2ox1 and PhGA2ox8 were amplified by PCR. And then analyzed the bioinformation of these sequences. The expression levels of PhGA2ox1 and PhGA2ox8 were analyzed using qPCR. Results: Bioinformatic analysis showed that PhGA2ox1 contained 981 bp and encoded a predicted protein of 326 amino acids with molecular weight of 36 871.3 and isoelectric point (PI) of 8.66; PhGA2ox8 contained 1 056 bp and encoded a predicted protein of 351 amino acids with molecular weight of 40 169.9 and PI of 6.91. Both PhGA2ox1 and PhGA2ox8 contained GA2ox conserved domains DIOX_N and 20G-Fell_Oxy, without obvious hydrophobic region, transmembrane domain and signal peptide. Phylogenetic analysis showed that the GA2ox of plant could be divided into three classes, PhGA2ox1 bolongs to Class I and PhGA2ox8 belongs to Class III. The qPCR. showed that the expression of PhGA2ox1 in different tissues and organs of P. heterophylla was constant, but the expression of PhGA2ox8 in root xylem was significantly higher than that in other tissues and organs (P < 0.05). Conclusion: The PhGA2ox1 and PhGA2ox8 genes are cloned for the first time, and provide a foundation for they may play an important role in the growth and development process of P. heterophylla.
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To provide guidance for certification, popularization and application of Pseudostellariae Radix new variety, the regional adaptation and stabilities of "Shitai No.1" were evaluated. The "Qian taizishen No.1" and "SB-C" varieties (strains) were used as the control varieties. The agronomic, medicinal material traits and medicine quality were used as evaluation index to compare the phenotypic difference of the three varieties (strains) in four planting areas. Compared to the control varieties, 10 agronomic traits of "Shitai No.1" had the smallest coefficient of variation among the 18 agronomic traits, and other 8 agronomic traits placed the middle level. Among 8 medicinal material traits and medicine quality indicators, the coefficient of variation of different regions of the extract content, pseudostellarin B content, the number of 50 g root tuber, the plant medicinal materials weight and weight of single root of "Shitai No.1" were the smallest compare to other varieties (strains). It could be divided into three groups based on the phenotypic difference of the three varieties (strains) in four planting areas. The "Shitai No.1" was classified as one group, while the "Qian taizishen No.1" and "SB-C" had cross clustering. The regional stability of several index about agronomic traits, medicinal material traits and medicine quality of "Shitai No.1" were better than that of the control varieties (strains). "Shitai No.1" was suitable for planting, popularization and application in the appropriate ecological areas of Guizhou province.
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As an important part of the market commodity circulation, the standard grade of Chinese traditional medicine commodity is very important to restrict the market order and guarantee the quality of the medicinal material. The State Council issuing the "protection and development of Chinese herbal medicine (2015-2020)" also make clear that the important task of improving the circulation of Chinese herbal medicine industry norms and the commodity specification standard of common traditional Chinese medicinal materials. However, as a large class of Chinese herbal medicines, the standard grade of the radix is more confused in the market circulation, and lack of a more reasonable study model in the development of the standard. Thus, this paper summarizes the research background, present situation and problems, and several key points of the commodity specification and grade standard in radix herbs. Then, the research model is introduced as an example of Pseudostellariae Radix, so as to provide technical support and reference for formulating commodity specifications and grades standard in other radix traditional Chinese medicinal materials.
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Abscisic acid 8'-hydroxylase was one of key enzymes genes in the metabolism of abscisic acid (ABA). Seven menbers of abscisic acid 8'-hydroxylase were identified from Pseudostellaria heterophylla transcriptome sequencing results by using sequence homology. The expression profiles of these genes were analyzed by transcriptome data. The coding sequence of ABA8ox1 was cloned and analyzed by informational technology. The full-length cDNA of ABA8ox1 was 1 401 bp,with 480 encoded amino acids. The predicated isoelectric point (pI) and relative molecular mass (MW) were 8.55 and 53 kDa,respectively. Transmembrane structure analysis showed that there were 21 amino acids in-side and 445 amino acids out-side. High level of transcripts can detect in bark of root and fibrous root. Multi-alignment and phylogenetic analysis both show that ABA8ox1 had a high similarity with the CYP707As from other plants,especially with AtCYP707A1 and AtCYP707A3 in Arabidopsis thaliana. These results lay a foundation for molecular mechanism of tuberous root expanding and response to adversity stress.
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OBJECTIVE: To establish a QTRAP UPLC-MS/MS method for simultaneous determination of thirteen nucleosides and nucleobases in Pseudostellariae Radix, and analyze their dynamic changes at different periods. METHODS: The separation was carried out on a Waters Atlantis T3 column (2.1 mm × 150 mm, 3 μm)eluted by a mobile phase of 5 mmol · mL-1 of ammonium acetate (B)-methanol(A) at a flow rate of 0.4 mL · min-1. The elution program is as follows. 0 -4.5 min, 3% -4% A; 4.5-8 min, 4% -18% A; 8 -10 min, 18%A; 10 -10.1 min, 18% -3% A; 10.1 -13 min, 3%A. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. RESULTS: The calibration curves of the 13 nucleosides and nucleobases showed good linearity (r >0.991 0) in the ranges of the tested concentrations, and the average recoveries were between 96.46% - 105.07%. The contents of cytidine, uridine, inosine, guanosine, and thymidine in Pseudostellariae Radix harvested at different periods showed some differences, and the contents were higher in mid-June. CONCLUSION: The method is simple, sensitive, accurate and credible, and provides the basis for exploring the quality forming mechanism of Pseudostellariae Radix medicinal materials and the suitable harvest time.
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Objective: To investigate the influence of different processing methods on the contents of nucleosides in Pseudostellariae Radix (PR). Methods: Q-TRAP-LC-MS/MS method was applied for the analysis on 13 kinds of nucleosides in PR with different processing methods. Results: The contents of cytidine, uridine, inosine, guanosine, and thymidine were higher in PR. The different processing methods of PR had certain influence on the contents of nucleosides. The amount of nucleosides in PR by oven drying was higher than that by drying in the sun. Conclusion: The paper investigates the influence of processing methods on chemical composition in PR, and provides the basis for establishing reasonable processing technology.