Standardized allergen extracts for allergen immunotherapy.

Nineteen U.S. allergen extracts were standardized by the U.S. Food and Drug Administration (FDA) between 1987 and 1998, including of two house-dust mites, short ragweed, cat hair and cat pelt, seven temperate and one southern grass, and six Hymenoptera venom preparations. Relevant literature was reviewed. For each allergen, a "representative" extract was established; the potency of each representative extract was determined by measurement of the total protein content (Hymenoptera venom), radial diffusion measurement of the dominant allergen (short ragweed and cat), or, if there was no dominant allergen, then by quantitative skin testing by using the ID50EAL (intradermal dilution for 50 mm sum of erythema determines the bioequivalent allergy units) method. In vitro tests were developed to allow the manufacturer to demonstrate that each lot of its extract was statistically identical, within defined limits, to the FDA reference extract. These tests included radial immunodiffusion, competitive enzyme-linked immunosorbent assay, and isoelectric focusing. The standardized extracts offer the advantage of consistent potency from lot to lot for each manufacturer and also from manufacturer to manufacturer, and assure the presence of recognized significant allergens within the extract. Therefore, standardized extracts offer improved safety and efficacy over their nonstandardized predecessors.

The Optimization of Extraction Process, Antioxidant, Whitening and Antibacterial Effects of Fengdan Peony Flavonoids.

Flavonoids have important biological activities, such as anti-inflammatory, antibacterial, antioxidant and whitening, which is a potential functional food raw material. However, the biological activity of Fengdan peony flavonoid is not particularly clear. Therefore, in this study, the peony flavonoid was extracted from Fengdan peony seed meal, and the antioxidant, antibacterial and whitening activities of the peony flavonoid were explored. The optimal extraction conditions were methanol concentration of 90%, solid-to-liquid ratio of 1:35 g:mL, temperature of 55 °C and time of 80 min; under these conditions, the yield of Fengdan peony flavonoid could reach 1.205 ± 0.019% (the ratio of the dry mass of rutin to the dry mass of peony seed meal). The clearance of Fengdan peony total flavonoids to 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, hydroxyl radical and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical could reach 75%, 70% and 97%, respectively. Fengdan peony flavonoid could inhibit the growth of the Gram-positive bacteria. The minimal inhibitory concentrations (MICs) of Fengdan peony flavonoid on S. aureus, B. anthracis, B. subtilis and C. perfringens were 0.0293 mg/mL, 0.1172 mg/mL, 0.2344 mg/mL and 7.500 mg/mL, respectively. The inhibition rate of Fengdan peony flavonoid on tyrosinase was 8.53-81.08%. This study intensely illustrated that the antioxidant, whitening and antibacterial activity of Fengdan peony total flavonoids were significant. Fengdan peony total flavonoids have a great possibility of being used as functional food materials.

Traditional uses, phytochemistry, pharmacology, toxicity and quality control of medicinal genus Aralia: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Aralia, which belongs to Araliaceae family, is mainly distributed in Asia, such as China, Japan and South Korea. It has a long medicinal history and is widely used in the treatment of various diseases, such as hepatitis, rheumatoid arthritis, bruises, lumps and carbuncles. AIM OF THE STUDY: The purpose of this review is to systematically evaluate the traditional uses, phytochemistry, pharmacology, toxicity and quality control of main medicinal plants of Aralia, discusses the application of ethnic medicine, modern scientific research and the relationship between them, and put forward some suggestions to promote the further development and utilization of Aralia. MATERIALS AND METHODS: The relevant information on Aralia was collected through electronic databases (PubMed, Web of Science, Science Direct, Springer, CNKI and Wanfang), Chinese herbal classics, Ph.D. and M.Sc. dissertations, Chinese Pharmacopoeia. Plant names were verified by "The Plant List" (http://www.theplantlist.org). The literature cited in this review can be traced back to 1878 to 2021. RESULTS: More than 290 chemical constituents have been isolated from the genus Aralia, including triterpenoid saponins, terpenoids, organic acids, flavonoids, polyacetylenes, phenylpropanoids and other constituents. Pharmacological studies have shown that the extracts and compounds of Aralia have a wide range of pharmacological activities, including anti-inflammation, analgesic, anti-tumor, liver protection, protection of cardiovascular and nervous system, regulating substance metabolism, antibacterial, antiviral and antioxidation. CONCLUSIONS: The genus Aralia is not only an excellent traditional herbal medicine, but also a source of bioactive molecules with good application prospects. However, the structure-activity relationship, in vivo activity and action mechanism of its bioactive components need to be further studied. In addition, more toxicological and quality control studies are essential to evaluate the efficacy and safety of Aralia as medicine.

Discrepancies in the German Pharmacopoeia procedure for quality control of Quillaja saponin extracts.

This study focused on the evaluation of Quillaja saponin extracts with the additional quality designation DAB-which means the abbreviation of the German Pharmacopoeia (Deutsches Arzneibuch). This label suggests that Quillaja saponin extracts marked in this way are of pharmacopoeial quality and thus stand out from other Quillaja saponin extracts. The DAB ninth edition listed Quillaia saponin as a reagent. According to DAB, its quality must be checked by thin-layer chromatography (TLC), and three closely spaced zones in a defined retention factor (Rf) interval specify the saponin reagent. All the Quillaja saponin extracts obtained from different manufacturers and labeled as DAB quality complied with the TLC test. However, the analysis with high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (HPLC-Q-ToF-MS) clearly showed additionally an intense peak pattern of Madhuca saponins in all measured samples. The TLC test for Mahua seed cake, which is the press residue from Madhuca longifolia, surprisingly showed the same three closely spaced zones in the defined Rf interval. The three zones could be identified as Mi-saponins from Madhuca after scraping and extracting them from the stationary phase of the TLC plate and subsequent measurement by HPLC-Q-ToF-MS. Therefore, the specification of the saponin reagent in DAB characterizes erroneously Madhuca saponins that are not listed as a saponin plant source for the saponin reagent.

Discovery of quality control ingredients in burdock root by combining anti-tumor effects and UHPLC-QqQ-MS/MS.

Burdock root is the root of Arctium lappa L., a plant of the Compositae family, which has the effects of dispersing wind and heat, detoxifying and reducing swelling. In order to better control the quality of burdock root, a screening study of quality control indicators was carried out. The current research combines biological activity evaluation with chemical analysis to screen and identify the biologically active compounds of burdock root as chemical components for the quality control of herbal medicine. The efficacy of 10 batches of ethanol extracts of burdock roots was evaluated by a tumor inhibition experiment in S180 tumor-bearing mice. The five main chemical components of these extracts were simultaneously quantitatively measured by ultra-high performance liquid chromatography combined with triple quadrupole mass spectrometry. Pearson correlation analysis was used to establish the relationship between these extracts' biological activity and chemical properties. The results showed that chlorogenic acid, caffeic acid and cynarin were positively correlated with the effect of inhibiting tumor growth, and further bioassays confirmed this conclusion. In conclusion, chlorogenic acid, caffeic acid and cynarin can be used as quality control markers for burdock root's antitumor effect.

Pentas longiflora Oliv. (Rubiaceae), a plant used in the treatment of Pityriasis Versicolor in Rwanda: Chemical composition and standardization of leaves and roots.

In Rwanda, the roots of Pentas longiflora Oliv. (Rubiaceae) have been used for a long time to treat Pityriasis versicolor. However, many people reported the use of leaves instead of roots. This research was conducted to compare the phytochemical composition and establish chromatographic methods for the standardization of roots and leaves extracts of P. longiflora. During this process, three new pentalongin glycosides (pentalonginoside A, pentalonginoside B, and pentalonginoside C) and two known glycosides of the same type (harounoside and clarinoside), as well as rutin, luteolin-7-rutinoside were isolated from methanol extract of leaves. In addition, pentalongin and psychorubrin, previously isolated from ethylacetate roots extract, were also identified in Pentas longiflora ethylacetate leaves extract. The presence of the antifungal compound pentalongin in leaves may explain the traditional use of leaves in the treatment of Pytiriasis versicolor. Furthermore, harounoside, psychorubrin, and pentalongin were selected as markers for HPLC fingerprints of MeOH extract. The accuracy and risk profile demonstrated the reliability of the validated method. In general, considerable variations of concentration in plant metabolites, including pentalongin, were observed between samples from different sites. The content in pentalongin (expressed as juglone) in collected samples ranged between 1.7 and 70.0 mg/100 g. The highest concentration (70.0 ± 17 mg/100 g) was registered in the cultivated samples from Mukoni. This important variation of pentalongin concentrations according to sampling sites, shows that in order to guarantee equivalent efficacy, finished products with P. longiflora should be standardized based on their pentalongin content.

A simple and effective method for the preparation of high-purity shikimic acid from chromatography wash effluent of Ginkgo biloba leaf extract by macroporous resin considering the effect of varying feed solution compositions.

OBJECTIVES: The present study investigated the feasibility of preparing high-purity shikimic acid (SA) from the chromatography wash effluent of Ginkgo biloba leaf extract by macroporous resin. METHODS: First, static/dynamic adsorption and desorption were conducted to screen out the optimal resin. Second, the key parameters of the chromatographic process were optimised with face-centred central composite design (CCD). Third, wash effluent indices were measured, different batches of wash effluent were used to prepare SA under the optimised parameters, and the effect of varying feed solution compositions on final products was investigated. KEY FINDINGS: It was found that the final purity and recovery rate of SA prepared with ADS-21 resin were not lower than 70 and 60%, respectively, when the purity of SA in the wash effluent was higher than 21.4%. The quality of the final product can be predicted based on the properties of wash effluent. CONCLUSIONS: The proposed method could not only provide a simple, green and promising approach for the large-scale purification of SA from wash effluent but also be used to develop process intermediate quality standards for other natural products.

Pharmacokinetics and metabolomics investigation of an orally modified formula of standardized Centella asiatica extract in healthy volunteers.

The formula of a standardized extract of Centella asiatica (ECa 233) was modified to improve its dissolution, with implications for pharmacokinetics and metabolomic profile. This study aimed to understand the resultant changes in disposition kinetics of ECa 233 and alterations to human metabolome after oral administration. This study was a two-sequence of dosages (250 and 500 mg), with an open-label phase I clinical trial. The modified formula was administered in single and multiple doses to twelve healthy Thai volunteers. The major parent compounds, madecassoside and asiaticoside, were rarely absorbed, instead undergoing biotransformation into active metabolites, madecassic acid and asiatic acid with possibility to be eliminated via fecal route. Increasing the dose of ECa 233 resulted in significantly greater plasma levels of those active metabolites, with accumulation of asiatic acid after multiple oral administration for seven days. Examining the impacts of accumulation behavior on metabolomics, the study traced changes in levels pre- and post-dose of five relevant human metabolites. Administration of ECa 233 was found to be significantly associated with an increase of choline, an endogenous metabolite with documented benefits for learning and memory. Therefore, ECa 233 may be useful in mitigating cognitive impairment, through its role in modulating human metabolites.

1H nuclear magnetic resonance-based metabolite profiling of guava leaf extract: an attempt to develop a prototype for standardization of plant extracts.

BACKGROUND: Herbal medicines are fast gaining popularity. However, their acceptability by modern practitioners is low which is often due to lack of standardization. Several approaches towards standardization of herbals have been employed. The current study attempted to recognize key peaks from 1H NMR spectra which together would comprise of a spectral fingerprint relating to efficacy of Psidium guajava (guava) leaf extract as an antidiarrhoeal when a number of unidentified active principles are involved. METHODS: Ninety samples of guava leaves were collected from three locations over three seasons. Hydroalcoholic (water and ethanol, 50:50) extracts of these samples were prepared and their 1H NMR spectra were acquired. Spectra were also obtained for quercetin, ferulic acid and gallic acid as standards. Eight bioassays reflecting different stages of diarrhoeal pathogenesis were undertaken and based on pre-decided cut-offs, the extracts were classified as 'good' or 'poor' extracts. The bioactivity data was then correlated with the 1H NMR profiles using Regression or Orthogonal Partial Least Square-Discriminant Analysis (OPLS-DA). RESULTS: OPLS-DA showed seasonal and regional segregation of extracts. Significant models were established for seven bioassays, namely those for anti-bacterial activity against Shigella flexneri and Vibrio cholerae, adherence of E. coli, invasion of E. coli and S. flexneri and production and binding of toxin produced by V. cholerae. It was observed that none of the extracts were good or bad across all the bioassays. The spectral analysis showed multiple peaks correlating with a particular activity. Based on NMR and LC-MS/MS, it was noted that the extracts contained quercetin, ferulic acid and gallic acid. However, they did not correlate with the peaks that segregated extracts with good and poor activity. CONCLUSIONS: The current study identified key peaks in 1H NMR spectra contributing to the anti-diarrhoeal activity of guava leaf extracts. The approach of using spectral fingerprinting employed in the present study can thus be used as a prototype towards standardization of plant extracts with respect to efficacy.

2D LC as a tool for standardization of Foenugraeci semen extracts containing compounds with anti-Helicobacter pylori activity.

The on-line heart-cutting two-dimensional liquid chromatography method with the use of a diode array detector and a mass spectrometer (LC-LC-DAD-ESI-MS) was established and validated for quantitation of C-glycosylflavones in fenugreek seeds (Foenugraeci semen, Trigonella foenum-graecum L.). The first- (1D) and second- (2D) dimensional separations were performed on Kinetex C-18 columns with different diameters, respectively, and gradient (1D) and isocratic elution (2D). Finally, 17 compounds were separated, 13 of which were quantified by 1D separation and 4 compounds by 2D separation. As a result, it was pointed out that fenugreek seeds of Polish origin can be considered as a rich source of C-glycosylflavones. Antibacterial activity against Helicobacter pylori of standardized 70% methanol extract from fenugreek seeds has been demonstrated, in contrast to the inactive aqueous extract. Anti-H. pylori activity of the 70% methanol extract can be related to a higher concentration of C-glycosylflavones. This is the first report on the bactericidal activity of vitexin, diosgenin, tigogenin and sarsasapogenin against H. pylori and the bacteriostatic activity of orientin against this bacterium.

The Untargeted Capability of NMR Helps Recognizing Nefarious Adulteration in Natural Products.

Curcuma longa (turmeric) has an extensive history of ethnomedical use for common ailments, and "curcumin"-containing dietary supplements (CDS) are a highly visible portion of today's self-medication market. Owing to raw material cost pressure, CDS products are affected by economically motivated, nefarious adulteration with synthetic curcumin ("syncumin"), possibly leading to unexpected toxicological issues due to "residual" impurities. Using a combination of targeted and untargeted (phyto)chemical analysis, this study investigated the botanical integrity of two commercial "turmeric" CDS with vitamin and other additives that were associated with reported clinical cases of hepatotoxicity. Analyzing multisolvent extracts of the CDS by 100% quantitative 1H NMR (qHNMR), alone and in combination with countercurrent separation (CCS), provided chemical fingerprints that allowed both the targeted identification and quantification of declared components and the untargeted recognition of adulteration. While confirming the presence of curcumin as a major constituent, the universal detection capability of NMR spectroscopy identification of significant residual impurities, including potentially toxic components. While the loss-free nature of CCS captured a wide polarity range of declared and unwanted chemical components, and also increased the dynamic range of the analysis, (q)HNMR determined their mass proportions and chemical constitutions. The results demonstrate that NMR spectroscopy can recognize undeclared constituents even if they represent only a fraction of the mass balance of a dietary supplement product. The chemical information associated with the missing 4.8% and 7.4% (m/m) in the two commercial samples, exhibiting an otherwise adequate curcumin content of 95.2% and 92.6%, respectively, pointed to a product integrity issue and adulteration with undeclared synthetic curcumin. Impurities from synthesis are most plausibly the cause of the observed adverse clinical effects. The study exemplifies how the simultaneously targeted and untargeted analytical principle of the 100% qHNMR method, performed with entry-level high-field instrumentation (400 MHz), can enhance the safety of dietary supplements by identifying adulterated, non-natural "natural" products.

Determination of Mogroside V in Luohanguo Extract for Daily Quality Control Operation Using Relative Molar Sensitivity to Single-Reference Caffeine.

Mogroside V is one of the characteristic and effective components of luohanguo extract, a food additive used as a sweetener in Japan as per Japan's Standards and Specifications for Food Additives (JSFA; 9th ed.). JSFA stipulates that the quantitative determination for mogroside V content in luohanguo extract applies HPLC using analytical standard mogroside V. However, no mogroside V reagents with proven purities are commercially available. Therefore the current JSFA determination method is not particularly suited for daily quality control operations involving luohanguo extract. In this study, we applied an alternative quantitative method using a single reference with relative molar sensitivity (RMS). It was possible to calculate the accurate RMS by an offline combination of 1H-quantitative NMR spectroscopy (1H-qNMR) and an HPLC/variable-wavelength detector (VWD). Using the RMS of mogroside V to a commercial certified reference material grade caffeine, the mogroside V contents in luohanguo extracts could be determined using HPLC/VWD without analytical standard mogroside V. There was no significant difference between the mogroside V contents in luohanguo extracts determined using the method employing single-reference caffeine with the RMS and using the JSFA method. The absolute calibration curve for the latter was prepared using an analytical standard mogroside V whose purity was determined by 1H-qNMR. These results demonstrate that our proposed method using a single reference with RMS is suitable for quantitative determination of mogroside V in luohanguo extract and can be used as an alternative method to the current assay method in JSFA.

Metabolomics of Withania somnifera (L.) Dunal: Advances and applications.

ETHNOPHARMACOLOGICAL RELEVANCE: Withania somnifera L. (Solanaceae), commonly known as Ashwagandha or Indian ginseng, is used in Ayurveda (Indian system of traditional medicine) for vitality, cardio-protection and treating other ailments, such as neurological disorders, gout, and skin diseases. AIM OF THE REVIEW: We present a critical overview of the information on the metabolomics of W. somnifera and highlight the significance of the technique for use in quality control of medicinal products. We have also pointed out the use of metabolomics to distinguish varieties and to identify best methods of cultivation, collection, as well as extraction. MATERIAL AND METHODS: The relevant information on medicinal value, phytochemical studies, metabolomics of W. somnifera, and their applications were collected from a rigorous electronic search through scientific databases, including Scopus, PubMed, Web of Science and Google Scholar. Structures of selected metabolites were from the PubChem. RESULTS: The pharmacological activities of W. somnifera were well documented. Roots are the most important parts of the plant used in Ayurvedic preparations. Stem and leaves also have a rich content of bioactive phytochemicals like steroidal lactones, alkaloids, and phenolic acids. Metabolomic studies revealed that metabolite profiles of W. somnifera depended on plant parts collected and the developmental stage of the plant, besides the season of sample collection and geographical location. The levels of withanolides were variable, depending on the morpho/chemotypes within the species of W. somnifera. Although studies on W. somnifera were initiated several years ago, the complexity of secondary metabolites was not realized due to the lack of adequate and fool-proof technology for phytochemical fingerprinting. Sophistications in chromatography coupled to mass spectrometry facilitated the discovery of several new metabolites. Mutually complementary techniques like LC-MS, GC-MS, HPTLC, and NMR were employed to obtain a comprehensive metabolomic profile. Subsequent data analyses and searches against spectral databases enabled the annotation of signals and dereplication of metabolites in several numbers without isolating them individually. CONCLUSIONS: The present review provides a critical update of metabolomic data and the diverse application of the technique. The identification of parameters for standardization and quality control of herbal products is essential to facilitate mandatory checks for the purity of formulation. Such studies would enable us to identify the best geographical location of plants and the time of collection. We recommend the use of metabolomic analysis of herbal products based on W. somnifera for quality control as well as the discovery of novel bioactive compounds.

A review of the ethnobotanical value, phytochemistry, pharmacology, toxicity and quality control of Tussilago farfara L. (coltsfoot).

ETHNOPHARMACOLOGICAL RELEVANCE: Tussilago farfara L. (commonly called coltsfoot), known as a vital folk medicine, have long been used to treat various respiratory disorders and consumed as a vegetable in many parts of the world since ancient times. AIM OF THE REVIEW: This review aims to provide a critical evaluation of the current knowledge on the ethnobotanical value, phytochemistry, pharmacology, toxicity and quality control of coltsfoot, thus provide a basis for further investigations. MATERIALS AND METHODS: A detailed literature search was obtained using various online search engines (e.g. Google Scholar, Web of Science, Science Direct, Baidu Scholar, PubMed and CNKI). Additional information was sourced from ethnobotanical literature focusing on Chinese and European flora. The plant synonyms were validated by the database 'The Plant List' (www.theplantlist.org). RESULTS: Coltsfoot has diverse uses in local and traditional medicine, but similarities have been noticed, specifically for relieving inflammatory conditions, respiratory and infectious diseases in humans. Regarding its pharmacological activities, many traditional uses of coltsfoot are supported by modern in vitro or in vivo pharmacological studies such as anti-inflammatory activities, neuro-protective activity, anti-diabetic, anti-oxidant activity. Quantitative analysis (e.g. GC-MS, UHPLC-MRMHR) indicated the presence of a rich (>150) pool of chemicals, including sesquiterpenes, phenolic acids, flavonoids, chromones, pyrrolizidine alkaloids (PAs) and others from its leaves and buds. In addition, adverse events have resulted from a collection of the wrong plant which contains PAs that became the subject of public concern attributed to their highly toxic. CONCLUSIONS: So far, remarkable progress has been witnessed in phytochemistry and pharmacology of coltsfoot. Thus, some traditional uses have been well supported and clarified by modern pharmacological studies. Discovery of therapeutic natural products and novel structures in plants for future clinical and experimental studies are still a growing interest. Furthermore, well-designed studies in vitro particularly in vivo are required to establish links between the traditional uses and bioactivities, as well as ensure safety before clinical use. In addition, the good botanical identification of coltsfoot and content of morphologically close species is a precondition for quality supervision and control. Moreover, strict quality control measures are required in the studies investigating any aspect of the pharmacology and chemistry of coltsfoot.

Effect of Standardized Hydroalcoholic Extract of Saffron Stamen on High Blood Pressure and Baroreflex Sensitivity in Anesthetized Rats.

OBJECTIVE: The stamen is a byproduct of saffron (Crocus sativus) flowers. Herein, its cardiovascular effects were evaluated on hypertension induced by angiotensin II (AngII) and NG-nitro-Larginine methyl ester (L-NAME), as well as baroreflex sensitivity (BRS). METHODS: Rats were randomly divided into 10 groups: 1) control, 2) AngII (50 ng/kg, i.v.), 3) losartan (10 mg/kg, i.p.) + AngII, 4) L-NAME (10 mg/kg, i.v.), 5) sodium nitroprusside (SNP) (50 mg/kg, i.p.) + L-NAME, 6, 7) saffron stamen extract (SS) (100 and 200 mg/kg, i.p.) + AngII and 8, 9) SS (100 and 200 mg/kg) + L-NAME, and 10) SS (200 mg/kg) + phenylephrine (Phen, i.v.). The treated rats first received two doses of SS, 30 min after the injection of L-NAME, AngII, and Phen in separate groups. The cardiovascular parameters were recorded by the PowerLab apparatus via an angiocatheter inserted into the femoral artery. The maximal changes (Δ) of mean arterial pressure (MAP), systolic blood pressure (SBP), and heart rate (HR) in the treated groups were compared with those of the hypertensive and control groups. The changes in MAP and HR induced by Phen were used for BRS evaluation. RESULTS: The SS extract did not significantly affect the basal cardiovascular parameters. The injection of AngII significantly increased the MAP and SBP (P<0.01-P<0.001) with no significant effect on the HR. The SS extract significantly attenuated the pressor effect induced by AngII (P<0.001). Increased MAP and SBP induced by L-NAME (P<0.001) were also significantly attenuated by the SS extract (P<0.01). The effect of SS extract on L-NAME was significantly higher than that of AngII (P<0.05). Moreover, BRS was significantly improved by the SS extract. CONCLUSION: Our findings provide evidence that the SS extract has anti-hypertensive effects that are probably mediated by an inhibitory effect on AngII, increasing nitric oxide production, or improving baroreflex sensitivity.

A comprehensive quality evaluation of Fuzi and its processed product through integration of UPLC-QTOF/MS combined MS/MS-based mass spectral molecular networking with multivariate statistical analysis and HPLC-MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Aconiti Lateralis Radix Praeparata (the Chinese name is Fuzi, FZ), the lateral or daughter root of Aconitum carmichaelii Debx. (Ranunculaceae), is a controversial traditional Chinese medicine (TCM) that is universally distributed and applied in many countries, such as China, Japan, Korea, and India. FZ can be used to treat various diseases, including rheumatic fever, rheumatism, painful joints, syncope, collapse, bronchial asthma, some endocrinal disorders, etc. However, quality control and assessment of FZ are challenging due to its obvious and high toxicological risks, and only its processed products are allowed to be used clinically according to the relative safety regulations. Consequently, it is necessary to analyze the whole chemical composition and the dynamic changes of FZ before and after processing. Addressing the changes in the chemical substance of raw and processed products is a way to reduce toxicity. AIM OF THE STUDY: In this article, the whole chemical composition of FZ is analyzed, the differences between raw and processed FZ are evaluated, and possible factors that influence the reduced toxicity of processed FZ are explained from the perspective of its chemical composition using qualitative and quantitative analysis methods. MATERIALS AND METHODS: A novel strategy of multiple data collection and processing based on ultra-performance liquid chromatography coupled with a quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) method in the positive ion mode, together with Global Natural Product Social Molecular Networking (GNPS) and multivariate statistical analysis, was established to systematically identify the chemical constituents of FZ and comprehensively investigate the chemical markers that can be used to differentiate FZ processed with vinegar and honey from its raw product. Combined with the qualitative analysis results, 12 components, including 8 chemical marker compounds and 4 toxicity components, were quantitatively analyzed by using high-performance liquid chromatography equipped with triple-quadrupole mass spectrometry (HPLC-MS/MS). RESULTS: Using the molecular networking (MN) analysis method, a total of 145 compounds were identified, of which 13 were identified using reference compounds. Seventy seven chemical markers were also detected between raw and processed FZ. The identification results of the chemical markers were also verified by orthogonal partial least squares discriminant analysis (OPLS-DA). The quantitative results indicated that the contents of 12 important components all decreased, especially diester-diterpenoid alkaloids (DDAs), after processing. CONCLUSION: The decrease of toxicity of FZ after processing is closely related to the changes in its chemical composition. The method developed in this study is a comprehensive analysis technique for quality assessment of FZ, and this study provides a useful and quick strategy to characterize chemical compounds of TCM and explore the different chemical markers between raw and processed Chinese herbal medicine.

Chemistry, safety, and regulatory considerations in the use of nitrite and nitrate from natural origin in meat products - Invited review.

Nitrite and nitrate have been traditionally used for the preservation of meat products because of the effective antimicrobial action of nitrite against Clostridium botulinum, the outgrowth of its spores as well as other bacteria. However, the use of nitrite and nitrate has been questioned in last half century due to the possible generation of N-nitrosamines through reaction of nitrite with secondary amines. Nitrite replacement strategies began in the 70s addressing these issues and instigated searches for natural alternatives to nitrate and nitrite, or for natural sources of nitrite and nitrate such as vegetable extracts. These alternatives have been considered by producers and consumers as an attractive practice even though they may also have some risks. This manuscript reviews and discusses the chemistry, safety, and regulatory considerations in the use of nitrite and nitrate from natural origin for the preservation of meat products.

A Two-Step Orthogonal Chromatographic Process for Purifying the Molecular Adjuvant QS-21 with High Purity and Yield.

QS-21 is a triterpene glycoside saponin found in the bark of the Chilean soap bark tree Quillaja saponaria. It is a highly potent vaccine adjuvant that is included in two approved vaccines and has shown promise in numerous other vaccine candidates in the research and clinical pipelines. One major hurdle to the widespread use of this adjuvant is the difficulty of obtaining it in high yield and purity. Previously reported purification approaches either showed suboptimal purity and/or yield, lacked efficiency, or had strict requirement on the composition of the starting material. Here, we report the development of a new two-step orthogonal chromatographic process, consisting of a polar reversed-phase (RP) chromatography step followed by a hydrophilic interaction chromatography (HILIC) step, for purifying QS-21 from a commercially available Quillaja saponaria bark extract with high yield and > 97% purity. This process makes available a simple and efficient method for obtaining highly pure QS-21 from saponin-enriched bark extract.

Metabolomics characterizes the metabolic changes of Lonicerae Japonicae Flos under different salt stresses.

Salt stress affects the metabolic homeostasis of medicinal plants. However, medicinal plants are sessile organisms that cannot escape from salt stress. They acclimatize themselves to the stress by reprogramming their metabolic pathways. Lonicerae Japonicae Flos (LJF) with strong antioxidant activity is commonly used in traditional Chinese medicine, tea, and beverage. Nevertheless, the variation of integrated metabolites in LJF under different salt stresses remains unclear. In this study, High Performance Liquid Chromatography tandem triple time-of-flight mass spectrometry (HPLC- triple TOF-MS/MS) coupled with multivariate statistical analysis was applied to comparatively investigate the metabolites changes in LJF under different salt stress (0, 100, 200, 300 mM NaCl). Total 47 differential metabolites were screened from 79 metabolites identified in LJF under different salt stress. Low salt-treated group (100 mM NaCl) appeared to be the best group in terms of relative contents (peak areas) of the wide variety in bioactive components. Additionally, the phenylpropanoid pathway, monoterpenoid biosynthesis, glycolysis, TCA cycle, and alkaloid biosynthesis were disturbed in all salt-stress LJF. The results showed that LJF metabolisms were dramatically induced under salt stress and the quality of LJF was better under low salt stress. The study provides novel insights into the quality assessment of LJF under salt stress and a beneficial framework of knowledge applied to improvement the medicinal value of LJF.

Endotoxin Contamination and Reaction Interfering Substances in the Plant Extract Library.

Endotoxin is an unintentional contaminant that has numerous activities and can affect various biological experiments using cells. In this study, we measured the endotoxin activity of samples from a plant extract library (PEL) and determined their degrees of contamination. Endotoxin was detected in approx. 48% (n = 139) and approx. 4% (n = 5) of field-collected and crude drug samples, respectively, and in concentrations >5.0 EU/mL in some samples. The concentrations of endotoxin that affect cells in vitro vary depending on the target cell type. Although the degree of contamination varied in the present study, it was considered to have little effect on the cell experiments. More than 150 PEL samples had problems with reaction courses or recovery rates of Limulus amoebocyte lysate (LAL) tests. In the LAL tests, using three plant extracts [Sanguisorba officinalis L. (Rosaceae), Oenothera biennis L. (Onagraceae), and Lythrum salicaria L. (Lythraceae)], the polyphenolic compounds in the plant extracts affected LAL test and their effects differed depending on the plant species. When the 16 single polyphenol compounds were added to the LAL tests, the compounds with caffeoyl and pyrogallol moieties were found to affect the LAL reaction course and recovery rate. Furthermore, none of the compounds had any effects at concentrations of 1 µM. Because the plant extracts contained analogs of various polyphenolic compounds, they were presumed to actually act synergistically. Our findings demonstrated that attention must be paid to the recovery rate and reaction process of LAL tests with samples containing polyphenolic compounds.