Common Methods of Extraction and Determination of Phytomelatonin in Plants.

The presence of melatonin in plants, called phytomelatonin, has gained great interest in recent years. The determination of phytomelatonin levels in plant extracts for both physiological and plant foodstuff studies requires sophisticated techniques due to the low endogenous levels of this indolic compound with hormonal nature. This chapter presents the most common and advanced techniques in the determination of phytomelatonin, with special emphasis on the techniques of extraction, cleaning, separation, detection, identification, and quantification. Multiple examples and recommendations are presented for a clear overview of the pros and cons of phytomelatonin determinations in plant tissues, seeds, and fruits, mainly.

An insight into recent updates on analytical techniques for bioactive alkaloids.

INTRODUCTION: Alkaloids represent a wide class of naturally existing nitrogen-containing organic compounds having diverse biological activities. They are primary bioactive substances extracted from diverse plant parts. Due to their diverse biological activities, they are frequently used as medicines. The alkaloids have diverse pharmacological impacts on the human body; alkaloids are used for prevention, treatment, and reduction of discomfort associated with chronic illnesses. As most alkaloids exist in plants in complex form, combined with numerous other natural plant components, it is essential to recognize and characterize these molecules using different analytical techniques. OBJECTIVES: We aimed to review the literature on the methods and protocols for the analysis of naturally occurring alkaloids. METHODS: We carried out a literature survey using the PubMed, Scopus, and Google Scholar databases and other relevant published materials. The keywords used in the searches were "alkaloids," "analytical methods," "HPLC method," "GC method," "electrochemical methods," and "bioanalytical methods," in various combinations. RESULTS: In this article, several classes of alkaloids are presented, along with their biological activities. Moreover, it includes a thorough explanation of chromatographic techniques, hyphenated techniques, electrochemical techniques, and current trending analytical methods utilized for the isolation, identification, and comprehensive characterization of alkaloids. CONCLUSIONS: The various analytical techniques play an important role in the identification as well as the characterization of various alkaloids from plants, plasma samples, and urine samples. The hyphenation of various chromatographic techniques with mass spectrometry and NMR spectroscopy plays a crucial role in the characterization of unknown compounds.

ß-Escin: An Updated Review of Its Analysis, Pharmacology, Pharmacokinetics, and Toxicity.

[Formula: see text]-Escin is an oleanane-type pentacyclic triterpenoid saponin extracted from the seeds of Aesculus hippocastanum (AH), which is more widely distributed. [Formula: see text]-Escin sodium has been approved by the American FDA for clinical usage. This paper is intended to summarize an updated and comprehensive review of the pharmacological activities, pharmacokinetic properties, toxicity, and analytical methods of [Formula: see text]-escin. Studies have shown that [Formula: see text]-escin has significant antitumor, antiviral, anti-inflammatory, and other activities alongside less adverse effects and higher safety than other compounds. The review shows that the pharmacological effects of [Formula: see text]-escin involve mechanisms such as ATM/[Formula: see text]H2AX, RhoA/Rock, GSK-3[Formula: see text]/[Formula: see text]-Catenin, HER2/HER3/Akt, and PI3K/Akt signaling pathways, and Cyclin A, p21[Formula: see text], survivin, Bcl-2, Mcl-1, Caspases, TGF-[Formula: see text], MMPs, and TNF-[Formula: see text] among other inflammatory factors. [Formula: see text]-Escin has significant cytotoxicity; the use of the chitosan/xanthan gum-based polyelectrolyte complexes PA1 and PC-11 to modify it not only to reduces its toxicity, but also improves its drug efficacy. Because of this, these compounds may become a new research hotspot. [Formula: see text]-Escin in vivo metabolism can be converted by the CYP1A2 enzyme in the intestinal flora to produce [Formula: see text]-escin, deacylated, deglycosylated, and 21[Formula: see text]-[Formula: see text]-crotonoyl-protoescin, and the binding rate of the plasma proteins is higher than 90%. These are mainly metabolized by the liver, kidneys, and other organs, and excreted in the form of urine and feces. The number of reports on the specific mediators of the metabolism of [Formula: see text]-escin and their mechanisms and metabolites is relatively small; furthermore, the results are vague. Therefore, a complete and in-depth exploration of the pharmacokinetic characteristics of [Formula: see text]-escin is needed to provide a more complete and effective theoretical reference for the study of its pharmacodynamic activity.

A systematic review and meta-analysis of validated analytical techniques for the determination of total selenium in foods and beverages.

The intricate balance between the beneficial and harmful effects of selenium (Se) intake means that its quantification in food needs to be done correctly. Therefore, in this review, we systematized 105 articles to identify the most studied methodologies, analytical techniques, and food matrices. Among the analytical techniques employed, inductively coupled plasma mass spectrometry (ICP-MS) (n = 29) emerged as the most commonly used method. The most prevalent hydrolysis methodology to digest Se in food matrices involved the use of nitric acid combined with ultrasound, which improved both the yield and digestion time. Optimal recovery values were achieved when total Se quantification accounted for the sum of Se(IV) and Se(VI) (94.4-99.4%) and for SeCys (88-96.5%). These findings are relevant for advancing methodological approaches, and their results emphasize the importance of developing alternative, faster, and lower-cost protocols for Se quantification in foods and beverages.

Sinapis Semen: A review on phytochemistry, pharmacology, toxicity, analytical methods and pharmacokinetics.

Sinapis Semen (SS), the dried mature seed of Sinapis alba L. and Brassica juncea (L.) Czern. et Coss., is one of the traditional Chinese medicinal materials with a wide range of pharmacological effects being used for asthma, cough and many other ailments. SS is also widely used in food agriculture, medicine and other industries in North America and South Asia. More recently, the research on SS has gradually intensified and increased. However, there is no systematic review of SS. In this review, through literature exploration and analysis, the research advance on phytochemistry, pharmacology, toxicity, analytical methods and pharmacokinetics of SS was aggregated initially. Total 144 compounds have been isolated and identified from SS. Among them, glucosinolates and their hydrolysates and volatile oils are the main active ingredients and important chemical classification markers. SS has a wide range of pharmacological effects, especially in cough suppressing, asthma calming, anti-inflammatory, neuroprotective, cardiovascular protective, inhibiting androgenic effects, anti-tumor, and skin permeation promoting effects. Sinapine and sinapic acid are the main active ingredients of SS for its medicinal effects. However, SS has a strong skin irritation, presumably related to the time of application, the method of processing, and original medicinal plants. This review will provide useful data for the follow-up research and safe and reasonable clinical application of SS.

Technical note: colorimetric methods for accurate determination of nutrient composition in beef cow colostrum and milk.

Our objectives were to develop colorimetric methods to accurately measure nutrient concentrations of beef cow colostrum and milk, to determine if the yield of colostrum from a single rear quarter is representative of complete collection of colostrum in beef cows, and to compare data from our developed colorimetric methods with Fourier transform infrared spectroscopy (FTIR) analysis to determine the accuracy of FTIR for beef cow colostrum and milk. In Exp. 1, colostral weight and volume of the most full rear quarter were compared with complete collection of colostrum from post-calving, unsuckled beef heifers. Both volume and weight had r2 = 0.85 (P < 0.001) between single-quarter and 4 quarter yields. In Exp. 2, colostrum (n = 35) and milk at d 35 (n = 42) and d 60 (n = 38) of lactation were collected from multiparous, fall-calving, crossbred beef cows. Subsamples were submitted for FTIR analysis and frozen for colorimetric analysis. Colorimetric analyses were developed for lactose, triglycerides (measure of fat), protein, and urea N. To validate method accuracy, spike recoveries were determined for lactose (96.8% average) and milk protein (100.1% average), triglyceride concentration was compared with fat concentration determined by the Mojonnier method (r2 ≥ 0.91, P < 0.001), and colostral or milk urea N was compared with serum urea N from the same sampling day (r2 ≥ 0.72, P < 0.001). Coefficients of determination between colorimetric methods and FTIR were determined for colostrum, d 35 milk, and d 60 milk. Colostral lactose concentration from FTIR was positively associated (r2 = 0.24, P = 0.01) with colorimetric analysis, but there was no relationship (r2 ≤ 0.09, P ≥ 0.14) between methods for colostral fat, protein, or urea N. Milk nutrient composition was positively associated for all nutrients measured at d 35 (r2 = 0.28 to 0.58, P < 0.001), and coefficients of determination strengthened for all nutrients measured at d 60 (r2 = 0.38 to 0.82, P < 0.001). In conclusion, colostrum yield of a single rear quarter can be used to indicate complete collection of colostrum for beef cows, and colorimetric methods developed have adequate accuracy for beef cow colostral and milk nutrient analysis. Based on our analyses, nutrient composition of beef cow colostrum was not accurately analyzed by FTIR. Accuracy of FTIR for beef cow milk varies with component and may be affected by the day of lactation.

The purpose was to develop laboratory methods to measure lactose, fat, protein, and urea nitrogen in beef cow colostrum (first milk) and milk and to validate single-quarter colostrum yield as a predictor of total colostrum yield. Additionally, new methods were compared with Fourier transform infrared spectroscopy (FTIR), which is the primary method of dairy cow milk nutrient analysis. New laboratory methods were determined to be accurate for beef cow colostrum and milk analysis, and single-quarter colostrum yield was successful in predicting total colostrum yield. Overall, our data suggest that beef cow colostrum cannot be accurately analyzed by FTIR instruments, and accuracy of FTIR for beef cow milk analysis varies with nutrients and may be affected by the day of lactation.

Stepwise Frontal Analysis Coupled with Affinity Chromatography: A Fast and Reliable Method for Potential Ligand Isolation and Evaluation from Mahuang-Fuzi-Xixin Decoction.

Mahuang-Fuzi-Xixin Decoction (MFXD) is widely used in the treatment of asthma, however, the functional components in the decoction targeting beta2-adrenoceptor (ß2 -AR) remain unclear. Herein, we immobilized the haloalkane dehalogenase (Halo)-tagged ß2 -AR on the 6-chlorocaproic acid-modified microspheres. Using the affinity stationary phase, the interactions of four ligands with the receptor were analyzed by stepwise frontal analysis. The association constants were (4.75±0.28)×104  M-1 for salbutamol, (2.93±0.15)×104  M-1 for terbutaline, (1.23±0.03)×104  M-1 for methoxyphenamine, (5.67±0.38)×104  M-1 for clorprenaline at high-affinity binding site, and (2.73±0.05)×103  M-1 at low-affinity binding site. These association constants showed the same rank order as the radioligand binding assay, demonstrating that immobilized ß2 -AR had capacity to screen bioactive compounds binding to the receptor while stepwise frontal analysis could predict their binding affinities. Application of the immobilized receptor in analysis of MFXD by chromatographic method revealed that ephedrine, aconifine, karakoline, and chasmanine were the bioactive compounds targeting ß2 -AR. Among them, ephedrine and chasmanine exhibited association constants of (2.94±0.02)×104 M-1 and (4.60±0.15)×104  M-1 to the receptor by stepwise frontal analysis. Molecular docking analysis demonstrated that ephedrine, chasmanine, and the other two compounds interact with ß2 -AR through the same pocket involving the key amino acids such as Asn312, Asp113, Phe289, Trp286, Tyr316, and Val114. As such, we reasoned that the four compounds dominate the therapeutic effect of MFXD against asthma through ß2 -AR mediating pathway. This work shed light on the potential of immobilized ß2 -AR for drug discovery and provided a valuable methodology for rapid screening.

Assessment of Undeclared Synthetic Drugs in Dietary Supplements in an Analytical View: A Comprehensive Review.

Dietary supplements have gained widespread attention globally as they are supposed to be healthier than synthetic pharmaceutical compounds with fewer side effects. Unlike common prescription drugs, dietary supplements are readily available to the general public. However, over the past few years, all kinds of legal and illegal drugs, have been detected in dietary supplements without labeling to defraud consumers, resulting in serious public health consequences. Due to the increasing number of drug frauds in dietary supplements, their consumption will undoubtedly risk consumers. Therefore; the importance of high-tech analytical methods in their assessment for controlling food quality seems undeniable. The current review examines the analytical techniques that focus on adulterating health products with undeclared pharmaceuticals, including registered and banned drugs. The present study referred to a survey from 2004 to 2021 based on articles in the Scopus database.

Illuminating Sub-Cellular Organelles by Small Molecule AIEgens.

Sub-cellular organelles play a critical role in a myriad biological phenomena. Consequently, organelle structures and functions are invariably highjacked in diverse diseases including metabolic disorders, aging, and cancer. Hence, illuminating organelle dynamics is crucial in understanding the diseased states as well as developing organelle-targeted next generation therapeutics. In this review, we outline the novel small molecules which show remarkable aggregation-induced emission (AIE) properties due to restriction in intramolecular motion (RIM). We outline the examples of small molecules developed to image organelles like mitochondria, endoplasmic reticulum (ER), Golgi, lysosomes, nucleus, cell membrane and lipid droplets. These AIEgens have tremendous potential for next-generation phototherapy.

Mechanism, indexes, methods, challenges, and perspectives of edible oil oxidation analysis.

Edible oils are indispensable food components, because they are used for cooking or frying. However, during processing, transport, storage, and consumption, edible oils are susceptible to oxidation, during which various primary and secondary oxidative products are generated. These products may reduce the nutritional value and safety of edible oils and even harm human health. Therefore, analyzing the oxidation of edible oil is essential to ensure the quality and safety of oil. Oxidation is a complex process with various oxidative products, and the content of these products can be evaluated by corresponding indexes. According to the structure and properties of the oxidative products, analytical methods have been employed to quantify these products to analyze the oxidation of oil. Combined with proper chemometric analytical methods, qualitative identification has been performed to discriminate oxidized and nonoxidized oils. Oxidative products are complex and diverse. Thus, proper indexes and analytical methods should be selected depending on specific research objectives. Expanding the mechanism of the correspondence between oxidative products and analytical methods is crucial. The underlying mechanism, conventional indexes, and applications of analytical methods are summarized in this review. The challenges and perspectives for future applications of several methods in determining oxidation are also discussed. This review may serve as a reference in the selection, establishment, and improvement of methods for analyzing the oxidation of edible oil.HighlightsThe mechanism of edible oil oxidation analysis was elaborated.Conventional oxidation indexes and their limited values were discussed.Analytical methods for the determination of oxidative products and qualitative identification of oxidized and non-oxidized oils were reviewed.

Applied Analytical Methods for Detecting Heavy Metals in Medicinal Plants.

For thousands of years, medicinal plants (MPs) have been one of the main sources of drugs worldwide. However, recently, heavy metal pollution has seriously affected the quality and safety of MPs. Consuming MPs polluted by heavy metals such as Pb, Hg, and Cu significantly threaten the health of consumers. To manage this situation, the levels of heavy metals in MPs must be controlled. In recent years, this field has attracted significant attention, but few researchers have systematically summarized various analytical methods. Therefore, it is necessary to investigate methods that can accurately and effectively detect the amount of heavy metals in MPs. Herein, some important analytical methods used to detect heavy metals in MPs and their applications have been introduced and summarized in detail. These include atomic absorption spectrometry, atomic fluorescence spectrometry, inductively coupled plasma mass spectrometry, inductively coupled plasma atomic emission spectrometry, X-ray fluorescence spectrometry, neutron activation analysis, and anodic stripping voltammetry. The characteristics of these methods were subsequently compared and analyzed. In addition, high-performance liquid chromatography, ultraviolet spectrophotometry, and disposable electrochemical sensors have also been used for heavy metal detection in MPs. To elucidate the systematic and comprehensive information, these methods have also been briefly introduced in this review.

Best Practice in the chemical characterisation of extracts used in pharmacological and toxicological research-The ConPhyMP-Guidelines.

Background: Research on medicinal plants and extracts derived from them differs from studies performed with single compounds. Extracts obtained from plants, algae, fungi, lichens or animals pose some unique challenges: they are multicomponent mixtures of active, partially active and inactive substances, and the activity is often not exerted on a single target. Their composition varies depending on the method of preparation and the plant materials used. This complexity and variability impact the reproducibility and interpretation of pharmacological, toxicological and clinical research. Objectives: This project develops best practice guidelines to ensure reproducibility and accurate interpretations of studies using medicinal plant extracts. The focus is on herbal extracts used in pharmacological, toxicological, and clinical/intervention research. Specifically, the consensus-based statement focuses on defining requirements for: 1) Describing the plant material/herbal substances, herbal extracts and herbal medicinal products used in these studies, and 2) Conducting and reporting the phytochemical analysis of the plant extracts used in these studies in a reproducible and transparent way. The process and methods: We developed the guidelines through the following process: 1) The distinction between the three main types of extracts (extract types A, B, and C), initially conceptualised by the lead author (MH), led the development of the project as such; 2) A survey among researchers of medicinal plants to gather global perspectives, opportunities, and overarching challenges faced in characterising medicinal plant extracts under different laboratory infrastructures. The survey responses were central to developing the guidelines and were reviewed by the core group; 3) A core group of 9 experts met monthly to develop the guidelines through a Delphi process; and. 4) The final draft guidelines, endorsed by the core group, were also distributed for feedback and approval to an extended advisory group of 20 experts, including many journal editors. Outcome: The primary outcome is the "Consensus statement on the Phytochemical Characterisation of Medicinal Plant extracts" (ConPhyMP) which defines the best practice for reporting the starting plant materials and the chemical methods recommended for defining the chemical compositions of the plant extracts used in such studies. The checklist is intended to be an orientation for authors in medicinal plant research as well as peer reviewers and editors assessing such research for publication.

Analytical methods for 25-hydroxyvitamin D: advantages and limitations of the existing assays.

Vitamin D plays an important role in the absorption of calcium and phosphorus, bone metabolism. Various analytical methods are being used to measure the circulating vitamin D levels in the blood. Among all vitamin D metabolites, 25 hydroxyvitamin (25(OH) D) is the most well-known member of the vitamin D family. It is available in two forms, viz. 25 hydroxyvitamin D3 (25(OH)D3) and 25 hydroxyvitamin D2(25(OH)D2). These are stable and available in ample amounts in serum and hence are the most commonly measured during estimation of vitamin D. Although, immunoassay was the most common analytical technique, it could not differentiate between 25(OH) D2 and 25(OH) D3. HPLC is another low-cost technique but lacks sufficient sensitivity for determining trace amounts of 25(OH) D2 and D3. Further, more advanced techniques like liquid chromatography-mass spectrometry have been developed to provide better specificity, higher degree of separation and identification of different forms of vitamin D. However, this technique is not very user friendly. Thus, standardization of these analytical methods in clinical and research laboratories is essential for developing and implementing evidence based clinical guidelines. This will also enable achievement of comparable results via different methods, based on diverse principles. In this manuscript, we have presented a comprehensive overview of the efforts for standardization in vitamin D assessment, along with latest advances in methods used for 25(OH) D2 and 25(OH) D3 analysis. Finally, a summary of clinical trials associated with vitamin D has been presented to discuss all the analytical methods employed for measuring 25(OH) D2 and D3.

Quality assessment of African herbal medicine: A systematic review and the way forward.

BACKGROUND: In Africa, herbalism supplements allopathic medicine's efforts to ensure Universal Health Coverage attainment. This review was conducted to identify and to summarise current literature on methodological approaches used for quality control of herbal medicines in Africa, to evaluate the gaps associated with existing strategies within context of best practices, and make recommendations for future improvements. METHODS: A systematic search was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Articles were screened and assessed for eligibility. RESULTS: 118 articles were included into the study. There was a high preference for impurity profiling tests (77%) indicating the prioritization for tests that guarantee safety despite the limited analytical resources available. Other classes of tests reported included identification tests (29%), physicochemical tests (18%), and content assays (12%). Although standard methods exist in preparing samples for impurity tests, different techniques were observed in different studies, and this could lead to differences in analytical outcomes. Content assays focused on single marker assessments, which may be inadequate to comprehensively assess the quality of products. CONCLUSION: This review provides knowledge of existing strengths and challenges for herbal medicine quality assessments in Africa. For future it is recommended to implement more studies on contaminants (e.g. mycotoxins) and pharmaceutical adulterants. The use of chemometrics to develop analytical methods should be promoted. Also, stakeholders in the medicine quality industry in Africa need to effectively collaborate to establish a well co-ordinated and harmonized system to provide a sustainable framework for the GACP and GMP guided production and quality assurance of herbal medicines.

On the Oxidative Valorization of Lignin to High-Value Chemicals: A Critical Review of Opportunities and Challenges.

The efficient valorization of lignin is crucial if we are to replace current petroleum-based feedstock and establish more sustainable and competitive lignocellulosic biorefineries. Pulp and paper mills and second-generation biorefineries produce large quantities of low-value technical lignin as a by-product, which is often combusted on-site for energy recovery. This Review focuses on the conversion of technical lignins by oxidative depolymerization employing heterogeneous catalysts. It scrutinizes the current literature describing the use of various heterogeneous catalysts in the oxidative depolymerization of lignin and includes a comparison of the methods, catalyst loadings, reaction media, and types of catalyst applied, as well as the reaction products and yields. Furthermore, current techniques for the determination of product yields and product recovery are discussed. Finally, challenges and suggestions for future approaches are outlined.

Prunella vulgaris L. - A Review of its Ethnopharmacology, Phytochemistry, Quality Control and Pharmacological Effects.

Prunella vulgaris L. (PVL) is dried fruit spike of Lamiacea plant Prunella vulgaris L., which is a perennial herb with medicinal and edible homology used for thousands of years. PVL is bitter, acrid, cold, and belongs to the liver and gallbladder meridians. It clears the liver and dissipate fire, improve vision, disperse swelling, and has satisfactory clinical therapeutic effects on many diseases such as photophobia, dizziness, scrofula, goiter, breast cancer. The collection of information and data related to PVL comes from literatures retrieved and collated from various online scientific databases (such as CNKI, VIP, PubMed, Web of Science, Research Gate, Science Database), ancient books of traditional chinese medicine (Encyclopedia of Traditional Chinese Medicine, Classics of Traditional Chinese Medicine, Dictionary of Traditional Chinese Medicine), and Doctoral and Master's Dissertations. Currently, the major chemical constituents isolated and identified from PVL are triterpenoids, steroids, flavonoids, phenylpropanoids, organic acids, volatile oils and polysaccharides. Modern pharmacological studies have shown that PVL has a wide range of pharmacological activities, including anti-inflammatory, anti-tumor, antibacterial and antiviral effects, as well as immune regulation, antihypertensive, hypoglycemic, lipid-lowering, antioxidant, free radical scavenging, liver protection, sedative and hypnotic effects. This paper reviewes the botany, ethnopharmacology, traditional application, phytochemistry, analytical methods, quality control, pharmacological effects of PVL. It can be used not only as medicine, but also gradually integrated into the "medicine and food homology" and "Chinese medicine health" boom. More importantly, it has great potential for drug resources development. This paper deeply discusses the shortcomings of current PVL research, and proposes corresponding solutions, in order to find a breakthrough point for PVL research in the future. At the same time, it is necessary to further strengthen the research on its medicinal chemistry, mechanism of action and clinical application efficacy in the future, and strive to extract, purify and synthesize effective components with high efficiency and low toxicity, so as to improve the safety and rationality of clinical medication.

Significant Discrepancies in "Functional Characterization of Undenatured Type II Collagen Supplements: Are They Interchangeable?": A Rejoinder.

In 2021, an article was published comparing two commercial products containing undenatured type II collagen: b-2Cool® (Bioiberica, S.A.U., Barcelona, Spain; designated CII-X, X1, and X2) and UC-II (Lonza Consumer Health Inc, USA; designated CII-Y, Y1, and Y2). However, we feel certain concerns remain where clarification may be warranted.

Phytochemistry, synthesis, analytical methods, pharmacological activity, and pharmacokinetics of loganin: A comprehensive review.

Iridoid glycosides (IGs) are found in many medicinal and edible plants, such as Gardenia jasminoides, Cistanche tubulosa, Eucommia ulmoides, Rehmanniae Radix, Lonicera japonica, and Cornus officinalis. Loganin, an IG, is one of the main active ingredient of Cornus officinalis Sieb. et Zucc., which approved as a medicinal and edible plant in China. Loganin has been widely concerned due to its extensive pharmacological effects, including anti-diabetic, antiinflammatory, neuroprotective, and anti-tumor activities, etc. Studies have shown that these underlying mechanisms include anti-oxidation, antiinflammation and anti-apoptosis by regulating a variety of signaling pathways, such as STAT3/NF-κB, JAK/STAT3, TLR4/NF-κB, PI3K/Akt, MCP-1/CCR2, and RAGE/Nox4/p65 NF-κB signaling pathways. In order to better understand the research status of loganin and promote its application in human health, this paper systematically summarized the phytochemistry, analysis methods, synthesis, pharmacological properties and related mechanisms, and pharmacokinetics based on the research in the past decades.

Analysis of pectolytic enzymes and Alternaria spp. in fresh dill, mustard seeds, onions, and vinegar, and their influence on the softening of pickled cucumbers.

Proteolytic enzymes or fungi have long been identified as causing softening of pickled cucumbers. As the softening of cucumbers occurs mainly in the pasteurized state, this study considers the hypothesis that vinegar and the added spices could be responsible for this softening by studying polygalacturonase (endo-/exo-PG), pectinesterase (PE), and pectin lyase, as well as Alternaria spp. found in the spices. Because of the high endo-PG activity found in dill, this spice emerged as a possible factor causing spoilage. Compared to dill, the enzyme activity in mustard seeds is eight times lower, and only low levels of enzymes or Alternaria spp. are present in onions or vinegar. Different harvest times and the associated degree of freshness of dill also played a crucial role regarding the endo-PG activity of up to 25.11 U/g (30°C, mature and very woody dill in late July) but of less than 9.00 U/g in fresh and soft green dill harvested in late June. A temperature of 80°C, held for 3 min, reduced the enzyme activity to less than 1.0 U/g. A final examination of cucumbers with a fixed quantity of mustard seeds, vinegar, and onions but with different variants of dill showed that the quantity of dill and the other ingredients added to the jars is not a potential factor leading to cucumber softening, which conflicts with the hypothesis of cucumber spoilage by vinegar and spices. PRACTICAL APPLICATION: This work provides insights into the activity of various pectolytic enzymes and the load of Alternaria spp. in different ingredients used for pickle production. Based on these data and additional pasteurization experiments, this paper evaluates the influence of dill, onions, mustard seeds, and vinegar on cucumber softening.

Recent Progress on Mass Spectrum Based Approaches for Absorption, Distribution, Metabolism, and Excretion Characterization of Traditional Chinese Medicine.

BACKGROUND: The absorption, distribution, metabolism, and excretion (ADME) of traditional Chinese medicine (TCM) components are closely related to their therapeutic efficacy, toxic effects, and drug interactions. Based on the study of the whole process of ADME in TCM, it is important to screen out the key pharmacokinetic index components (pharmacokinetics PK/toxicokinetics TK makers), which can be beneficial for their clinical application or drug development. Although the detection of traditional small molecular drug's in vivo ADME process can be achieved by radioisotope methods, this approach might not be useful for the case of TCM. In detail, it is very difficult to label and trace each component in complex Chinese medicine, while it is also difficult to accurately follow the position of tracer in the whole in vivo process. In short, it is a tough task to obtain the ADME information of Chinese medicine, especially in the case of a clinical study. METHODS: We searched several scientific databases, including Pubmed, ACS, ScienceDirect, Springer, Wiley, etc., by using "Chinese medicine" and "in vivo metabolism" as keywords. By summarizing the current reports as well as our recent progress in this field, this review aims to summarize current research methods and strategies for ADME study of TCM based on high-resolution mass spectrometry-based data acquisition and data mining technology which is an important approach but has not been systematically reviewed. RESULTS: With the development of various hybrid tandem high-resolution mass spectrometry (Q-TOF, LTQ FT, Q-Exactive), liquid chromatography-high resolution mass spectrometry (LC-HRMS) has become the mainstream method in studying ADME process of TCM. This review aims to comprehensively summarize current research technologies and strategies based on high-resolution mass spectrometry, with emphasis on the following three aspects: (1) comprehensive and automatic acquisition technologies for the analysis of in vivo TCM components (i.e., BEDDA); (2) quick and comprehensive identification techniques for analyzing in vivo chemical substances and metabolites of TCM (i.e., PATBS or metabolomic analysis); (3) efficient correlation determination between in vivo or in vitro compounds and their metabolic transformation (i.e., MTSF). CONCLUSION: To the best of our knowledge, this is a pioneering review for systematically summarizing the analysis methods and strategies of ADME in TCM, which can help understand the whole ADME process, therapeutic molecular basis, or toxic substances of TCM. Furthermore, this review can also provide a feasible strategy to screen out PK/TK markers of TCM, while this information can be helpful to elucidate the pharmacodynamics or toxicity mechanisms of Chinese medicines and be useful in their future drug development. At the same time, we also hope that this review can provide ideas for further improvement of TCM analysis methods and help rational clinical use of TCM and the development of new drugs.