Glycyrrhizic Acid Content Research Articles

Coupling effects of irrigation amount and fertilization rate on growth and bioactive components of four-year-old licorice (Glycyrrhiza uralensis Fisch) in arid regions of Xinjiang

Water scarcity, over-fertilization, and improper crop management practices severely limit the sustainable cultivation of licorice (Glycyrrhiza uralensis Fisch) in the arid regions of Xinjiang. To elucidate the impacts of integrated water and fertilizer management on the growth characteristics and bioactive components (glycyrrhizic acid and liquiritin) of four-year-old licorice plants, a comprehensive four-year field experiment was conducted from 2019 to 2022.The experiment included four irrigation levels (W1: 2500 m³/ha, W2: 4000 m³/ha, W3: 5500 m³/ha, W4: 7000 m³/ha) and four fertilization rates (F1: 305 kg/ha, F2: 610 kg/ha, F3: 915 kg/ha, F4: 1220 kg/ha), following a completely randomized design. Results indicated that both irrigation and fertilization significantly influenced plant height, root length, root weight, root diameter, leaf area index, and root-to-shoot ratio. The optimal growth characteristics were observed under the W2F2 treatment. The contents of glycyrrhizic acid and liquiritin varied significantly among different water and fertilizer treatments, with the highest levels observed under the W2F2 treatment. Excessive irrigation (W4) and over-fertilization (F4) led to a decrease in these bioactive components. A comprehensive evaluation of the growth characteristics and bioactive components revealed that the ideal irrigation and fertilization parameters were 4000 m³/ha and 610 kg/ha, respectively. These parameters optimized plant development and bioactive component accumulation while ensuring efficient resource use. This study provides scientific evidence for optimizing irrigation and fertilization strategies to enhance licorice yield in arid regions, thereby supporting sustainable agricultural practices and improving economic benefits.

The interaction effect of water deficit stress and seaweed extract on phytochemical characteristics and antioxidant activity of licorice (Glycyrrhiza glabra L.).

With increasing drought stress due to climate change and water scarcity, the agricultural sector has sought innovative strategies to mitigate the detrimental effects on crop productivity. One approach that has received significant attention is the use of fertilizers and biostimulants as potential means of alleviating drought stress. In this study, five different irrigation levels including 100% (control), 80% (slight stress), 60% (mild stress), 40% (moderate stress), and 20% (severe stress) of field capacity (FC) and seaweed extract (SWE) at three concentrations (0, 5, and 10 g/L) were applied to the pots containing one-year-old licorice (Glycyrrhiza glabra L.) plants in a factorial completely randomized design experiment with three replications for eight weeks. The glycyrrhizic acid content increased with water stress intensity without the application of SWE until severe (20% FC) water stress treatment. The application of 10 g/L SWE under 100% FC led to a significant increase in the glycyrrhizic acid value (32.5±0.889 mg/g DW) compared with non-SWE application (30.0±1.040 mg/g DW). The maximum glabridin content (0.270±0.010 mg/g DW) was obtained under irrigation of 20% field capacity with 10 g/L SWE application. In addition, the activity of the all studied enzymes such as APX (ascorbate peroxidase), CAT (catalase), POD (peroxidase), and SOD (superoxide dismutase) were boosted by increasing the water stress levels. The use of SWE further enhanced the increase of some of these metabolites and enzymes, which, in turn, helped the plant to tolerate stress conditions through the scavenging of more ROS (Reactive oxygen species), wherein for this purpose, the SWE 10 g/L was more effective than other concentration. The plants efficiently eliminated ROS driven from drought stress by both non-enzymatic and enzymatic systems.

La (NO3)3 substantially fortified Glycyrrhiza uralensis’s resilience against salt stress by interconnected pathways

The taproot of Glycyrrhiza uralensis is globally appreciated for its medicinal and commercial value and is one of the most popular medicinal plants. With the decline of wild G. uralensis resources, cultivated G. uralensis has become a key method to ensure supply. However, soil salinization poses challenges to G. uralensis cultivation and affects the yield and quality of it. In this study, the inhibitory effects of NaCl and Na2SO4 on yield and quality of G. uralensis were comprehensively evaluated in a three-year large-scale pot experiment, and the alleviating effects of supplementation with lanthanum nitrate (La (NO3)3) on G. uralensis were further evaluated under salt stress. The findings indicate that La (NO3)3 significantly strengthened the plant's salt tolerance by enhancing photosynthetic capacity, osmolyte accumulation, antioxidant defenses, and cellular balance of ions, which led to a substantial increase in root biomass and accumulation of major medicinal components. In comparison to the NaCl-stress treatment, the 0.75 M La (NO3)3 + NaCl treatment resulted in a 20% and 34% increase in taproot length and biomass, respectively, alongside a 52% and 43% rise in glycyrrhizic acid and glycyrrhizin content, respectively. Similar improvements were observed with 0.75 M La (NO3)3 + Na2SO4 treatment, which increased root length and biomass by 14% and 26%, respectively, and glycyrrhizic acid and glycyrrhizin content by 40% and 38%, respectively. The combined showed that application of La (NO3)3 not only significantly improved the salt resilience of G. uralensis, but also had a more pronounced alleviation of growth inhibition induced by NaCl compared to Na2SO4 stress except in the gas exchange parameters and root growth. This study provides a scientific basis for high-yield and high-quality cultivation of G. uralensis in saline soils and a new approach for other medicinal plants to improve their salt tolerance.Graphical

Quantification of Glycyrrhizic Acid in FLEGMEN-SIP Capsules Using High-Performance Liquid Chromatography for CNS Disorders

Background: Neurological diseases affect over one billion people globally, presenting a significant health burden, especially for vulnerable populations. The FLEGMEN-SIP capsule was developed as a treatment option for central nervous system disorders, containing glycyrrhizic acid as its primary active ingredient. This study aims to quantify the glycyrrhizic acid content in FLEGMEN-SIP capsules using high-performance liquid chromatography (HPLC) to ensure compliance with therapeutic standards. Methods: The study utilized HPLC to quantify glycyrrhizic acid in FLEGMEN-SIP capsules. The capsules contain a dry extract derived from Phlomis regelii, Leonurus turkestanicus, Mentha piperita, and Glycyrrhiza glabra, with glycyrrhizic acid comprising 1.39% of the dry extract. Samples were prepared by dissolving 0.050 g of the drug in the mobile phase and filtered using a 0.45 µm membrane filter. The HPLC system included a Zorbax column and UV detection at 254 nm. The analysis was repeated five times for accuracy. Results: The glycyrrhizic acid content in FLEGMEN-SIP capsules was determined to be 0.55%, surpassing the minimum regulatory requirement of 0.4%. The chromatographic method demonstrated high reproducibility and reliability, confirming the quality and therapeutic efficacy of the capsules. Conclusion: The study validated the use of HPLC as an effective method for quantifying glycyrrhizic acid in FLEGMEN-SIP capsules. The results confirm the capsules meet regulatory standards, ensuring their suitability for treating central nervous system disorders.

Influence of Habitat and Effects of Salt Stress on Biochemical and Physiological Parameters of Glycyrrhiza uralensis.

The seeds of Glycyrrhiza uralensis Fisch. used for cultivating are primarily sourced from wild populations. However, the types of habitats where wild G. uralensis grow are diverse. We studied the effects of salinity on the growth, antioxidant capacity, and photosynthetic physiology of two-month-old licorice seedlings from different habitats to evaluate their salt tolerance. With the increasing NaCl concentration, compared with non-salinized habitats, seedlings originating from seeds collected from salinized habitats showed milder inhibition in root biomass and root volume. Also, the crown diameter increased more significantly. Activities of superoxide dismutase, catalase, and peroxidase are higher. Correspondingly, the electrolyte leakage rate of the leaves is low. Their leaves had a higher photoprotection capacity and potential maximum photochemical efficiency of PSII. Net photosynthetic rate, transpiration rate, and stomatal conductance showed less inhibition under 4 and 6 g/kg NaCl treatment. The content of glycyrrhizic acid and glycyrrhetinic acid in their roots was significantly increased under 2 g/kg NaCl treatment and was significantly higher than that of seedlings from non-salinized habitats under the same NaCl treatment. In conclusion, seeds from salinized habitats show improved tolerance to salt stress at the seedling stage, which is attributed to their superior phenotypic adaptability, strong antioxidant, and especially high light protection ability.

Glycerin as an alternative solvent for the extraction of glycyrrhizic acid from the roots of Glycyrrhiza glabra

Licorice root extracts are one of the most common and widely used plant extracts. Due to the high content of biologically active substances and pronounced properties, they are used in various industries – in medicine, pharmacology, cosmetics and food industries. The underground part of Licorice contains more than 100 different chemical substances, the most biologically active of which are flavonoids and triterpene saponins. The most significant saponin is glycyrrhizic acid. Science knows many ways to obtain licorice extract, and the research is still being actively carried out to find even more effective, inexpensive and accessible methods.The purpose of the research was to investigate the feasibility and potential of glycerol as a solvent for the extraction of glycyrrhizic acid from the roots of Glycyrrhiza glabra.The methods. Isolation of glycyrrhizic acid from the roots of Glycyrrhiza glabra was carried out using the maceration method. Glycerrin and the classic extractant ethanol were tested as an extractant for comparative analysis. Determination of glycyrrhizic acid content in licorice glabra roots was carried out using differential spectrophotometry.The Results. Optimal conditions for the extraction of glycyrrhizic acid from the roots of Glycyrrhiza glabra were selected. The maximum content of glycyrrhizic acid was observed in the sample prepared at a water-glycerol ratio of 3:2.The scientific novelty of the work lies in the study of the possibility of using glycerol as a solvent for extraction and the selection of optimal conditions for the extraction of glycyrrhizic acid from the roots of Glycyrrhiza glabra in order to increase the level of extraction of the target substance.

Introducing the glycyrrhizic acid and glabridin rich genotypes from the cultivated Iranian licorice (Glycyrrhiza glabra L.) populations to exploit in production systems

Currently, the stable, uniform, and highly efficient production of raw materials for pharmaceutical companies has received special attention. To meet these criteria and reduce harvesting pressure on the natural habitats of licorice (Glycyrrhiza glabra L.), cultivation of this valuable plant is inevitable. In the present study, to introduce the glycyrrhizic acid (GA)- and glabridin-rich genotypes from cultivated Iranian licorice, forty genotypes from eight high-potential wild populations were cultivated and evaluated under the same environmental conditions. The GA content varied from 5.00 ± 0.04 mg/g DW (TF2 genotype) to 23.13 ± 0.02 mg/g DW (I5 genotype). The highest and lowest glabridin content were found in the K2 (0.72 ± 0.021 mg/g DW) and M5 (0.02 ± 0.002 mg/g DW) genotypes, respectively. The rutin content in the leaves of the studied genotypes varied from 1.27 ± 0.02 mg/g DW in E4 to 3.24 ± 0.02 mg/g DW in BO5 genotypes. The genotypes from the Ilam population were characterized by higher vegetative growth and yield traits in the aerial parts and roots. The average root dry yield was 2.44 tons per hectare (t/ha) among the studied genotypes and a genotype from Ilam (I5) yielded the maximum value (3.08 ± 0.034 t/ha). The highest coefficient of variation among the genotypes was observed for leaf width (CV = 34.9%). The GA and glabridin-rich genotypes introduced in this study can be used in the future breeding programs to release new bred licorice cultivars.

Effects of Crude Extract of Glycyrrhiza Radix and Atractylodes macrocephala on Immune and Antioxidant Capacity of SPF White Leghorn Chickens in an Oxidative Stress Model.

The natural edible characteristics of Chinese herbs have led more and more people to study them as an alternative product to antibiotics. In this study, crude extracts of Glycyrrhiza radix and Atractylodes macrocephala (abbreviated as GRAM) with glycyrrhizic acid content not less than 0.2 mg/g were selected to evaluate the effects of GRAM on the immune and antioxidant capacity of model animals. Thirty 21-day-old male Leghorn chickens were weighed and randomly assigned to one of three groups of ten animals each. The treatments comprised a control group (CON), in which saline was injected at day 31, day 33, and day 35, an LPS-treated group (LPS), in which LPS (0.5 mg/kg of BW) was injected at day 31, day 33, and day 35, and finally a GRAM and LPS-treated group, (G-L) in which a GRAM-treated diet (at GRAM 2 g/kg) was fed from day 21 to day 35 with LPS injection (0.5 mg/kg of BW) at day 31, day 33, and day 35. The results of diarrhea grade and serum antioxidant measurement showed that the LPS group had obvious diarrhea symptoms, serum ROS and MDA were significantly increased, and T-AOC was significantly decreased. The oxidative stress model of LPS was successfully established. The results of immune and antioxidant indexes showed that feeding GRAM significantly decreased levels of the pro-inflammatory factors TNF-α, IL-1β, and IL-6 (p < 0.05) and significantly increased levels of the anti-inflammatory factors IL-4 and IL-10 and levels of the antioxidant enzymes GSH-Px and CAT (p < 0.05). GRAM resisted the influence of LPS on ileum morphology, liver, and immune organs and maintained normal index values for ileum morphology, liver, and immune organs. In summary, this study confirmed the antidiarrheal effect of GRAM, which improved the immune and antioxidant capacity of model animals by regulating inflammatory cytokine levels and antioxidant enzyme activity in poultry.

Investigating the effect of processing conditions on the content of glycyrrhizic acid in licorice (Radix et Rhizoma Glycyrrhizae)

Licorice is the root of Glycyrrhiza uralensis Fisch or Glycyrrhizae inflata L. or Glycyrrhiziae glabra L. It was a familiar medicinal herb in traditional medicine, which was commonly used in two different forms raw licorice (Sinh Cam thao) and honey-fried licorice (Chich Cam thao), with completely different effects. The processing enhanced the impact on the Spleen and Stomach organs and tonified Qi but reduced the bioactivities of clearing heat, antitussive, and detoxifying. The purpose of this study was to propose a processing method that minimizes the degradation of the main bioactive compound, glycyrrhizic acid in licorice. Honey-fired licorice was processed according to the guidance of Circular 30/2017/TT-BYT of the Vietnamese Ministry of Health. The frying conditions such as frying temperature, honey soaking time, the ratio of honey to licorice, and the ratio of water to honey were investigated in order. The content of glycyrrhizic acid was determined according to the process of Vietnamese Pharmacopoeia V. Glycyrrhizic acid was least decomposed when processing honey-fired licorice under the following conditions: the frying temperature of 260°C, the honey soaking time of 1 hour, the ratio of honey to licorice was 1:10, and the ratio of water to honey was 1:1. The content of glycyrrhizic acid in licorice was influenced by the frying process, in which temperature was the most significant factor. Honey was an ingredient that altered the effects of licorice in traditional medicine, however, the presence of honey also contributed to the degradation of glycyrrhizic acid in this medicinal herb.

Investigating the effect of processing conditions on the content of glycyrrhizic acid in licorice (Radix et Rhizoma Glycyrrhizae)

Licorice is the root of Glycyrrhiza uralensis Fisch or Glycyrrhizae inflata L. or Glycyrrhiziae glabra L. It was a familiar medicinal herb in traditional medicine, which was commonly used in two different forms raw licorice (Sinh Cam thao) and honey-fried licorice (Chich Cam thao), with completely different effects. The processing enhanced the impact on the Spleen and Stomach organs and tonified Qi but reduced the bioactivities of clearing heat, antitussive, and detoxifying. The purpose of this study was to propose a processing method that minimizes the degradation of the main bioactive compound, glycyrrhizic acid in licorice. Honey-fired licorice was processed according to the guidance of Circular 30/2017/TT-BYT of the Vietnamese Ministry of Health. The frying conditions such as frying temperature, honey soaking time, the ratio of honey to licorice, and the ratio of water to honey were investigated in order. The content of glycyrrhizic acid was determined according to the process of Vietnamese Pharmacopoeia V. Glycyrrhizic acid was least decomposed when processing honey-fired licorice under the following conditions: the frying temperature of 260°C, the honey soaking time of 1 hour, the ratio of honey to licorice was 1:10, and the ratio of water to honey was 1:1. The content of glycyrrhizic acid in licorice was influenced by the frying process, in which temperature was the most significant factor. Honey was an ingredient that altered the effects of licorice in traditional medicine, however, the presence of honey also contributed to the degradation of glycyrrhizic acid in this medicinal herb.

Effects of ultrasonic power on drying kinetics and product quality of licorice extract during ultrasound-assisted vacuum drying.

Licorice extract is an important raw material for food additives and medicine. The quality of licorice extract is dictated by the drying process. The commonly used drying methods of licorice extract are not efficient in obtaining high-quality products so alternative techniques need to be developed and researched. In this study, ultrasound-assisted vacuum drying (UAVD) was first utilized to improve drying efficiency and produce a higher-quality product. The changes in water mobility of licorice extract during drying were characterized using low-field nuclear magnetic resonance. In addition, the effects of ultrasonic power on the drying dynamics, the contents of liquiritin and glycyrrhizic acid, the antioxidant capacity and the microstructure formation of licorice extract during the whole drying process were investigated. The drying times for licorice extract to reach equilibrium moisture content were reduced by 9.09-69.70% with UAVD at 40-200 W compared with that without ultrasonic treatment (0 W). Moreover, the proportions of bound water and semi-bound water in fresh concentrate were 3.75% and 96.25%. It was also found that high ultrasonic power promoted the flow of water and the formation of porous structure in licorice extract, which led to the improvement of drying efficiency. The contents of liquiritin (2.444%) and glycyrrhizic acid (6.514%) were retained to a large degree in the dried product at an ultrasonic power of 80 W. The DPPH inhibition rate of UAVD samples with different ultrasonic powers ranged from 84.07 ± 0.46% to 90.65 ± 0.22%. UAVD has the advantages of high efficiency and low energy consumption, which may be an alternative technology for vacuum drying widely used in industry. © 2024 Society of Chemical Industry.

A low dose of daily licorice intake affects renin, aldosterone, and home blood pressure in a randomized crossover trial

BackgroundLicorice, through the effects of glycyrrhizic acid (GA), raises blood pressure (BP). The World Health Organization has suggested that 100 mg GA/d would be unlikely to cause adverse effects, but of 13 previously published studies none have been randomized and controlled and independently quantified the GA content. ObjectiveOur aim was to analyze the effects on home BP of a daily licorice intake containing 100 mg GA. MethodsHealthy volunteers were randomly assigned to start with either licorice or a control product in a nonblinded, 2 × 2 crossover study. Home BP was measured daily, and blood samples were collected at the end of each 2-wk period. ResultsThere were 28 participants and no dropouts. The median age was 24.0 y (interquartile range 22.8–27.0 y). During the licorice compared with control intake period, the systolic home BP increased [mean difference: 3.1 mm Hg (95% confidence interval [CI]: 0.8, 5.4 mm Hg) compared with −0.3 mm Hg (95% CI: −1.8, 1.3 mm Hg); P = 0.018] and renin and aldosterone were suppressed [mean change: −30.0% (95% CI: −56.7%, −3.3%) compared with 15.8% (95% CI: −12.8%, 44.4%); P = 0.003; and −45.1% (95% CI: −61.5%, −28.7%) compared with 8.2% (95% CI: −14.7%, 31.1%); P <0.001, respectively]. In the quartile of participants with the most pronounced suppression of renin and aldosterone, N-terminal prohormone of brain natriuretic peptide concentration increased during the licorice compared with control period [mean change: 204.1% (95% CI: −11.6%, 419.7%) compared with 72.4% (95% CI: −52.2%, 197.1%); P = 0.016]. ConclusionsWe found licorice to be more potent than previously known, with significant increases in BP, after a daily intake of only 100 mg GA. Thus, the safe limit of intake of this substance might need to be reconsidered.This trial was registered at clinicaltrials.gov as NCT05661721 (https://clinicaltrials.gov/study/NCT05661721).

Nutrient distribution in various tissues of Licorice (Glycyrrhiza glabra L.) and the influence of soil fertility on the levels of its bioactive compounds

This study aimed to optimize the production of licorice (Glycyrrhiza glabra L.) in field conditions using two licorice ecotypes and various types of nutrient sources. Licorice yield, leaf and root nutrient content, and its bioactive compounds were assessed. Mycorrhiza inoculation resulted in the highest rhizome dry weight in the Lalehzar (101.3 g) and Kermanshah (395.6 g) ecotypes in the first and second years, respectively. Rhizome dry weight was higher than the roots in both ecotypes. Two years old Kermanshah ecotype exhibited the highest root dry weight by using urea. The harvest index increased in Lalehzar ecotype by application of vermicompost in the first year and cow manure coupled with plant growth-promoting bacteria (PGPB) in the second year. Nitrogen content in leaves and roots significantly increased in the Kermanshah ecotype using cow manure and PGPB. Application of mycorrhiza (in Lalehzar ecotype) and cow manure combined with urea (in Kermanshah ecotype) led to leaf phosphorus (P) content improvement. The highest root P content in Lalehzar and Kermanshah ecotypes achieved by using cow manure and urea, respectively. Fertilization by urea and mycorrhiza led to higher leaf potassium (K) content in Kermanshah and Lalehzar, respectively, while urea yielded the highest root K content in the Kermanshah ecotype. Root glycyrrhizic acid (GA) content peaked in the Kermanshah ecotype with cow manure and PGPB (138 and 372.8 mg g-1 DW in the first and second years, respectively). Rhizome GA content performed better in the Lalehzar ecotype using cow manure and PGPB (191.34 mg g-1 DW, first year) and in the Kermanshah ecotype with cow manure and urea (239.62 mg g-1 DW, second year). Fertilizers had not significant effect on glabridin (GL) content. Notably, GA and GL exhibited a negative correlation with potassium and nitrogen. This study highlights the importance of balanced nutrient management for licorice cultivation.

Elucidating the active ingredients and potential anti-inflammatory mechanism of Qingwei Huanglian Wan

Qingwei Huanglian Wan (QHW) is a traditional Chinese medicine used for treating inflammation. As the compositions of QHW preparations are complex and varied, it has proven difficult to identify the active ingredients in QHW and control the quality. In this study, high-performance liquid chromatography (HPLC) fingerprints, network pharmacology, molecular docking, and the Griess method were used to screen for and verify the potential active ingredients of QHW. First, sixteen batches of QHW were studied as two groups using multivariate statistical analysis. Thirteen peaks were detected in the HPLC results to establish a fingerprint similarity model, and six chemical constituents (phellodendrine hydrochloride, geniposide, berberine hydrochloride, baicalin, wogonoside, and glycyrrhizic acid) were identified. Among these six constituents, four components (glycyrrhizic acid, berberine hydrochloride, baicalin, and wogonoside) were considered as potential active ingredients. Second, network pharmacology, molecular docking, and the Griess method were used to further elucidate the potential active ingredients. There was no significant difference in the NO content of the active ingredients administration group (dose converted into the medium-dose group) and medium-dose QHW administration group. Then, the contents of berberine hydrochloride, baicalin, wogonoside, and glycyrrhizic acid were determined using the HPLC method. This study provides a comprehensive and reliable strategy for the quality control of QHW preparations and identifies potential active ingredients in QHW. Our methodology may also be useful for studying other traditional Chinese medicines.

A quality-comprehensive-evaluation-index-based model for evaluating traditional Chinese medicine quality

BackgroundEvaluating traditional Chinese medicine (TCM) quality is a powerful method to ensure TCM safety. TCM quality evaluation methods primarily include characterization evaluations and separate physical, chemical, and biological evaluations; however, these approaches have limitations. Nevertheless, researchers have recently integrated evaluation methods, advancing the emergence of frontier research tools, such as TCM quality markers (Q-markers). These studies are largely based on biological activity, with weak correlations between the quality indices and quality. However, these TCM quality indices focus on the individual efficacies of single bioactive components and, therefore, do not accurately represent the TCM quality. Conventionally, provenance, place of origin, preparation, and processing are the key attributes influencing TCM quality. In this study, we identified TCM-attribute-based quality indices and developed a comprehensive multiweighted multi-index-based TCM quality composite evaluation index (QCEI) for grading TCM quality.MethodsThe area of origin, number of growth years, and harvest season are considered key TCM quality attributes. In this study, licorice was the model TCM to investigate the quality indicators associated with key factors that are considered to influence TCM quality using multivariate statistical analysis, identify biological-evaluation-based pharmacological activity indicators by network pharmacology, establish real quality indicators, and develop a QCEI-based model for grading TCM quality using a machine learning model. Finally, to determine whether different licorice quality grades differently reduced the inflammatory response, TNF-α and IL-1β levels were measured in RAW 264.7 cells using ELISA analysis.ResultsThe 21 quality indices are suitable candidates for establishing a method for grading licorice quality. A computer model was established using SVM analysis to predict the TCM quality composite evaluation index (TCM QCEI). The tenfold cross validation accuracy was 90.26%. Licorice diameter; total flavonoid content; similarities of HPLC chromatogram fingerprints recorded at 250 and 330 nm; contents of liquiritin apioside, liquiritin, glycyrrhizic acid, and liquiritigenin; and pharmacological activity quality index were identified as the key indices for constructing the model for evaluating licorice quality and determining which model contribution rates were proportionally weighted in the model. The ELISA analysis results preliminarily suggest that the inflammatory responses were likely better reduced by premium-grade than by first-class licorice.ConclusionsIn the present study, traditional sensory characterization and modern standardized processes based on production process and pharmacological efficacy evaluation were integrated for use in the assessment of TCM quality. Multidimensional quality evaluation indices were integrated with a machine learning model to identify key quality indices and their corresponding weight coefficients, to establish a multiweighted multi-index and comprehensive quality index, and to construct a QCEI-based model for grading TCM quality. Our results could facilitate and guide the development of TCM quality control research.

COMT, CRTZ, and F3′H regulate glycyrrhizic acid biosynthesis in Glycyrrhiza uralensis hairy roots

Glycyrrhiza uralensis Fisch. is prescribed as one of the original plants of licorice in Chinese Pharmacopoeia. This herbal medicine possesses numerous pharmacological activities and has been used in clinic in China since ancient times. Glycyrrhizic acid (GA) is a triterpenoid compound isolated from G. uralensis. It is often used as one of the marker components for the medicinal quality of the herb. Previously, we showed that the expression levels of three genes in G. uralensis were inversely correlated with the content of GA, including the caffeic acid 3-O-methyltransferase gene (COMT), the β-carotene 3-hydroxylase gene (CRTZ), and the flavonoid 3′-monooxygenase gene (F3′H). In this study, the main aim is to determine the roles of the three genes on GA production through gene knockout and overexpression in G. uralensis hairy roots. We observed that neither knockout nor overexpression of any of the genes affected the viability of the transgenic hairy roots, indicating that these genes are not essential for survival of hairy roots. Compared with the wild type and negative control hairy roots, GA content was significantly lower in hairy roots overexpressing COMT, CRTZ, or F3′H, but higher in those with anyone of the genes knocked out. Our findings demonstrate that the three genes, COMT, CRTZ, and F3′H, all negatively regulate the GA biosynthesis.

Effects of Different Drying Methods on Drying Characteristics and Quality of Glycyrrhiza uralensis (Licorice).

Large amounts of waste result from licorice mold rot; moreover, prompt drying directly influences product quality and value. This study compared various glycyrrhiza drying methods (Hot air drying (HAD), infrared combined hot air drying (IR-HAD), vacuum freeze drying (VFD), microwave vacuum drying (MVD), and vacuum pulsation drying (VPD)) that are used in the processing of traditional Chinese medicine. To investigate the effects of various drying methods on the drying characteristics and internal quality of licorice slices, their color, browning, total phenol, total flavonoid, and active components (liquiritin and glycyrrhizic acid) were chosen as qualitative and quantitative evaluation indices. Our results revealed that VFD had the longest drying time, but it could effectively maintain the contents of total phenol, total flavonoid, and liquiritin and glycyrrhizic acid. The results also showed that VFD samples had the best color and the lowest degree of browning, followed by HAD, IR-HAD, and VPD. We think that VFD is the best approach to ensure that licorice is dry.

Separation, characterization and anti-psoriasis effect of self-assembled nanoparticles from Shaoyao Gancao Decoction

This study aims to separate and characterize self-assembled nanoparticles(SAN) from Shaoyao Gancao Decoction(SGD) and determine the content of active compounds. Further, we aimed to observe the therapeutic effect of SGD-SAN on imiquimod-induced psoriasis in mice. The separation of SGD was performed by dialysis, and the separation process was optimized by single factor experiment. The SGD-SAN isolated under the optimal process was characterized, and the content of gallic acid, albiflorin, paeoniflorin, liquiritin, isoliquiritin apioside, isoliquiritin, and glycyrrhizic acid in each part of SGD was determined by HPLC. In the animal experiment, mice were assigned into a normal group, a model group, a methotrexate group(0.001 g·kg~(-1)), and SGD, SGD sediment, SGD dialysate, and SGD-SAN groups of different doses(1, 2, and 4 g·kg~(-1)) respectively. The psoriasis grade of mice was evaluated based on the pathological changes of skin lesions, the content of inflammatory cytokines, organ index and other indicators. The results showed that SAN obtained by centrifugation at 13 000 r·min~(-1) for 30 min was stable after dialysis for 4 times, which were uniform spherical nanoparticles with the particle size of(164.43±1.34) nm, the polydispersity index of(0.28±0.05), and the Zeta potential of(-12.35±0.80) mV. The active compound content accounted for more than 70% of SGD. Compared with the model group, SAN and SGD decreased the skin lesion score, spleen index, and inflammatory cytokine levels(P&lt;0.05 or P&lt;0.01) and alleviated the skin thickening and infiltration of inflammatory cells. However, the sediment group and the dialysate group had no obvious effect. SGD showed a good therapeutic effect on imiquimod-induced psoriasis in mice, and SAN demonstrated the effect equivalent to SGD in a dose-dependent manner. Therefore, we conclude that the SAN formed during decocting is the main active form of SGD, which can lower the levels of inflammatory cytokines, promote the normal differentiation of keratinocytes, and reduce the infiltration of inflammatory cells in the treatment of psoriasis lesions in mice.

Improving drought tolerance and glycyrrhizin content of licorice plant by EPS-producing cyanobacteria

We evaluated the effects of two selected EPS-producing cyanobacterial strains for microbe-dependent drought tolerance in native licorice plant (Glycyrrhiza glabra) and addressed the mechanisms, which might be improved through these interactions. These bacteria were inoculated on the soils in a form of single strain (Anabaena sphaerica or Nostoc pruniforme) or two strains (Anabaena + Nostoc) in greenhouse conditions. The growth parameters, physiological and metabolical responses among treatments were investigated through assaying the antioxidants and glycyrrhizin contents in the plants grown in both normal condition and moderate water stress condition (FC 50%). The reduction of the ratio of root/shoot in response to drought (FC 50%) was correlated with amount of specific metabolites production, including glycyrrhizic acid (GA), glabridin (GB), liquiritin (LQ) and liquiritigenin (LG), especially in soil treated by Anabaena + Nostoc (P < 0.05). In water-deficient conditions, accumulation of GA in plants treared by Anabaena + Nostoc was boosted by 64% and 69% compared to Anabaena and the control, respectively. Besides, a significant increase in catalase activity (25%) was perceived in this treatment after exposure to drought stress. These results highlight the potential role of enzymatic antioxidants and the increase of GA content in responsing to the drought tolerance in licorice plant. Our findings suggest that using EPS-producing strains of Cyanobacteria is an efficient treatment to improve the drought tolerance in cultivation of medicinal plant in soil of arid and semi-arid regions.

Analysis on quality value transmitting of Qinggusan reference sample of classical prescription based on quantitative fingerprint technology

Qinggusan is the 69th prescription in the first batch of "Catalogue of Ancient Chinese Classic Formulas". In modern clinical practice， Qinggusan is mainly used to treat noninfectious fever. However， because few studies on Qinggusan reference samples and their quality value transfer are available， the development and promotion of its compound preparations are restricted. Therefore， establishing an accurate and comprehensive quality control method to clarify the critical quality attributes of Qinggusan reference samples is of great importance. In this study， 15 batches of Qinggusan reference samples were processed to determine the range of their dry extract ratios. Quantitative high-performance liquid chromatography （HPLC） fingerprint analysis was performed using a Waters Symmetry Shield RP18 column （250 mm×4.6 mm， 5 μm） with acetonitrile-0.1% （v/v） formic acid aqueous solution as the mobile phase in gradient elution mode. The flow rate was 1.0 mL/min， the column temperature was 30 ℃， and the detection wavelength was 254 nm. The HPLC fingerprints of the Qinggusan reference samples were established under these conditions to evaluate their similarity. The established method was systematically validated and found to demonstrate good precision， repeatability， and sample stability. Subsequently， characteristic peaks were identified and attributed by HPLC-quadrupole-time-of-flight-mass spectrometry （HPLC-Q-TOF-MS） analysis. MS was performed in electrospray ionization mode， the data were collected in both positive- and negative-ion modes， and the detection range was m/z 50-2000. The contents and transfer rate ranges of the index components， namely， gentiopicrin， mangiferin， picroside Ⅱ， picroside Ⅰ， and glycyrrhizic acid， were determined to analyze the quality value transfer of the samples. The results demonstrated that the dry extract rate of the 15 batches of Qinggusan reference samples ranged from 24.10% to 26.88% and that their fingerprint similarities were generally greater than 0.95. Twelve common peaks were identified by reference identification， literature comparison， and high-resolution MS analysis. Twelve compounds， including six iridoid glycosides， two flavonoids， one alkaloid， one triterpenoid saponin， and two others. Among them， L-picein， androsin， picroside Ⅳ， picroside Ⅱ and picroside Ⅰ were from Picrorhizae Rhizoma， loganin acid， swertiamarin and gentiopicrin were from Gentianae Macrophyllae Radix， neomangiferin and mangiferin were from Anemarrhenae Rhizoma， dichotomine B was from Stellariae Radix， and glycyrrhizic acid was from Glycyrrhizae Radix et Rhizoma. The five key components presented good linear relationships in their respective linear ranges， and all correlation coefficients were higher than 0.999. The relative standard deviations （RSDs） of precision， stability， and repeatability were less than 1.3%. The average recoveries varied between 95.92% and 102.5%， with RSDs less than 3.9%； these values meet the requirements defined in the 2020 edition of the Chinese Pharmacopoeia. The contents of gentiopicrin， mangiferin， picroside Ⅱ， picroside Ⅰ， and glycyrrhizic acid in the 15 batches of reference samples were in the range of 17.92-27.55， 1.83-4.42， 23.08-36.44， 8.43-15.04， and 0.94-2.39 mg/g， respectively， and their transfer rates from the decoction pieces to the reference samples were 47.91%-63.95%， 22.96%-59.39%， 60.82%-77.82%， 64.25%-99.53%， and 15.30%-39.30%， respectively. In this study， the chemical components of Qinggusan reference samples were comprehensively identified and their quality value transfer was studied through the combination of HPLC fingerprinting and MS. Clarification of the critical quality attributes of Qinggusan reference samples could provide a basis for the quality control of Qinggusan compound preparations.