A guide to mycetisms: A toxicological and preventive overview.

Fungi are often considered a delicacy and are primarily cultivated and harvested, although numerous species are responsible for intoxication due to toxin content. Foodborne diseases are a significant public health concern, causing approximately 420 000 deaths and 600 million morbidities yearly, of which mushroom poisoning is one of the leading causes. Epidemiological data on non-cultivated mushroom poisoning in individual countries are often unrepresentative, as intoxication rarely requires emergency intervention. On the other hand, the lack of specialist knowledge among medical personnel about the toxicological manifestations of mushroom consumption may result in ineffective therapeutic interventions. This work aims to provide an easy-to-consult and wide-ranging tool useful for better understanding the variability of mushroom intoxications, the associated symptoms, and the main treatments for the most severe cases, given the absence of a complete species mapping tool toxic. Moreover, we establish an effective collection network that describes the incidence of mushroom poisonings by reporting the species and associated toxicological manifestations for each case. In conclusion, we highlight the need to establish appropriate primary prevention interventions, such as training the affected population and increasing consultancy relationships between mycological experts and specialised healthcare personnel.

We propose a review of the literature that describes the main syndromes resulting from the consumption of toxic fungal species, reporting symptoms and clinical manifestations, latency times and, where possible, diagnostic tools for recognising the species involved and interventions to be carried out.

Cold Alkali-Extractable Antioxidative Polysaccharide from Russula pseudocyanoxantha (Agaricomycetes), a Novel Mushroom, Stimulates Immune Responses in RAW264.7 Cells by Regulating the TLR/NF-κB Pathway.

In our previous study, we have established Russula pseudocyanoxantha as a unique species, playing a crucial role in indigenous diets through ages. The research also brought attention to bioactive potential of polysaccharide fraction extracted from the unexplored food using hot water. However, residue of the conventional process still contains therapeutic biopolymers that could further be utilized for pharmacological purposes instead of being discarded. Therefore, the current study aims to valorize the solid remnants, contributing to a deeper understanding of the novel taxon. Subsequently, the leftover was treated with cold alkali, leading to the preparation of a high-yield fraction (RP-CAP). Chemical characterization through FT-IR, GC-MS, HPTLC, and spectroscopy demonstrated presence of several monomers in the carbohydrate backbone, predominantly composed of ß-glucan. Furthermore, GPC chromatogram indicated presence of a homogeneous polymer with molecular weight of ~ 129.28 kDa. Subsequently, potent antioxidant activity was noted in terms of radical scavenging (O2·-, OH·, DPPH· and ABTS·+), chelating ability, reducing power and total antioxidant activity where EC50 values ranged from 472-3600 µg/mL. Strong immune-boosting effect was also evident, as the biopolymers stimulated murine macrophage cell proliferation, phagocytic activity, pseudopod formation, and NO as well as ROS synthesis particularly at the concentration of 100 µg/mL. In-depth analysis through RT-PCR revealed that the fraction stimulated synthesis of several inflammatory mediators, elucidating the mode of action through TLR/ NF-κB pathway. Therefore, the findings collectively suggest that RP-CAP possesses great potential to serve as a healthimproving component in functional food and pharmaceutical sectors.

Comprehensive Review of Functional and Nutraceutical Properties of Craterellus cornucopioides (L.) Pers.

Nutraceuticals represent an emerging and dynamic scientific field due to their important potential in integrated healthcare through nutritional and medicinal approaches that interact and complement each other mutually. In an attempt to find new sources for such preparations, the present research focuses on the species Craterellus cornucopioides (L.) Pers. (Cantharellaceae), also known as the black trumpet. This wild mushroom species is renowned for its culinary excellence and unique taste and is used especially in a dehydrated state. However, beyond its gastronomic value, recent scientific investigations have revealed its potential as a source of bioactive compounds with pharmaceutical and therapeutic significance. Our study aimed, therefore, to review the current data regarding the morphology, chemical profile, and medicinal potential of the black trumpet mushroom, highlighting its unique attributes. By conducting a comprehensive literature analysis, this paper contributes to the broader understanding of this remarkable fungal species as a potential functional food and its promising applications in the field of therapeutics.

Research Progress of Bioactive Components in Sanghuangporus spp.

The species in Sanghuangporus are a group of edible mushrooms with a long history of oral use in East Asia as a health-improvement method. They should be classified under the genus Sanghuangporus rather than mistakenly in Phellinus or Inonotus. The major components in this genus consist of polysaccharides, polyphenols, triterpenoids, and flavonoids, all of which exist in the fruiting bodies and mycelia. For extraction, studies have shown methods using hot water, ethanol, DES solvent, and alkaline, followed by purification methods including traditional anion column, Sevag solution, macroporous resin, and magnetic polymers. Proven by modern medical technology, these components possess promising anti-inflammatory, antioxidative, antitumor, and immunoregulation effects; additionally, they have health-improving effects including pulmonary protection, hypoglycemic properties, sleep improvement, gout mitigation, antiaging, neuroprotection, and muscle-strengthening abilities. Several toxicity studies have revealed their safety and recommend a dose of 1 g/kg for mice. As a newly emerged concept, functional food can provide not only life-sustaining nutrients but also some health-improving effects. In conclusion, we substantiate Sanghuang as a functional food by comprehensively presenting information on extraction and purification methods, component medical and structural properties, and nontoxicity, hoping to benefit the development of Sanghuang species as a group of functional food.

Extraction, structure and antioxidant activity of the polysaccharides from morels (Morchella spp.): A review.

Morels (Morchella spp.), which are cultivated only in a few regions of the world, are edible mushrooms known for their various properties including antioxidation, immune regulation, antiinflammation, and antitumor effects. Polysaccharides from Morchella are principally responsible for its antioxidant activity. This paper reviews the extraction, purification, structural analysis and antioxidant activity of Morchella polysaccharides (MPs), providing updated research progress. Meanwhile, the structural-property relationships of MPs were further discussed. In addition, based on in vitro and in vivo studies, the major factors responsible for the antioxidant activity of MPs were summarized including scavenging free radicals, reduction capacity, inhibitory lipid peroxidation activity, regulating the signal transduction pathway, reducing the production of ROS and NO, etc. Finally, we hope that our research can provide a reference for further research and development of MPs.

Food and human health applications of edible mushroom by-products.

Mushroom waste can account for up to 50% of the total mushroom mass. Spent mushroom substrate, misshapen mushrooms, and mushroom stems are examples of mushroom byproducts. In ancient cultures, fungi were prized for their medicinal properties. Aqueous extracts containing high levels of ß-glucans as functional components capable of providing prebiotic polysaccharides and improved texture to foods have been widely used and new methods have been tested to improve extraction yields. Similarly, the addition of insoluble polysaccharides controls the glycemic index, counteracting the effects of increasingly high-calorie diets. Numerous studies support these benefits in vitro, but evidence in vivo is scarce. Nonetheless, many authors have created a variety of functional foods, ranging from yogurt to noodles. In this review, we focus on the pharmacological properties of edible mushroom by-products, and the possible risks derived from its consumption. By incorporating these by-products into human or animal feed formulations, mushroom producers will be able to fully optimize crop use and pave the way for the industry to move toward a zero-waste paradigm.

Colorimetric assay for α-amanitin based on inhibition of carbon dots/AuNPs nanoenzyme activity.

Accidental ingestion of poisonous mushrooms leading to poisoning is a global issue. The most important and lethal toxin causing mushroom poisoning is α-amanitin, with a lethal dose of about 0.1 mg kg-1. Rapid detection of wild mushrooms before consumption or rapid identification of toxins after poisoning can effectively reduce the occurrence of fatalities. This study established a method for detecting α-amanitin using carbon dots/AuNPs nanoenzymes (D-Glu-CDs/AuNPs) with robust peroxidase-like activity. This nanoenzyme was prepared employing glucose carbon dots and sodium citrate as reducing and stabilizing agents, respectively. It could oxidize the substrate TMB (tetramethylbenzidine) to produce blue o-TMB. When α-amanitin specifically bound to the active site of the nanoenzyme, a resultant decrease was observed in catalytic activity and the absorbance value at 652 nm. The regression equation Y = -0.06083x + 0.9643, with an R2 value of 0.996, was obtained. The limit of detection was determined to be 48.03 ng mL-1, and the recoveries in urine ranged from 91.2% to 97.6%. This method enabled the visualization of α-amanitin, and the whole detection process was completed within 20 min. The approach holds promise for the quantitative and qualitative determination of α-amanitin in urine samples.

New autophagy-modulating lanostane-type triterpenoids from a hallucinogenic poisonous mushroom Gymnopilus orientispectabilis.

Gymnopilus orientispectabilis, also known as "big laughter mushroom," is a hallucinogenic poisonous mushroom that causes excessive laughter upon ingestion. From the fruiting bodies of G. orientispectabilis, eight lanostane-type triterpenoids (1-8), including seven novel compounds: gymnojunols A-G (2-8), were isolated. The chemical structures of these new compounds (2-8) were determined by analyzing their 1D and 2D NMR spectra and HR-EISMS, and their absolute configurations were unambiguously assigned by quantum chemical ECD calculations and a computational method coupled with a statistical procedure (DP4+). Upon evaluating autophagic activity, compounds 2, 6, and 7 increased LC3B-II levels in HeLa cells to a similar extent as bafilomycin, an autophagy inhibitor. In contrast, compound 8 decreased the levels of both LC3B-I and LC3B-II, and a similar effect was observed following treatment with rapamycin, an autophagy inducer. Our findings provide experimental evidence for new potential autophagy modulators in the hallucinogenic poisonous mushroom G. orientispectabilis.

The effect of casing and gypsum on the yield and psychoactive tryptamine content of Psilocybe cubensis (Earle) Singer.

Psychedelic fungi have experienced a surge in interest in recent years. Most notably, the fungal secondary metabolite psilocybin has shown tremendous promise in the treatment of various psychiatric disorders. The mushroom species that produce this molecule are poorly understood. Here we sought to examine for the first time, the response of a psilocybin-producing species Psilocybe cubensis to casing (peat moss and vermiculite) and supplementation with gypsum (calcium sulfate dihydrate), two common practices in commercial mushroom cultivation. Mycelial samples of genetically authenticated P. cubensis were used to inoculate popcorn grain bags. The fully colonized bags of popcorn grain (0.15 kg) were transferred to bins of 0.85 kg pasteurized horse manure, with or without 1 cm thick layer of casing and/or 5 % gypsum. Our results indicate that the use of a casing layer significantly increases the biological efficiency (161.5 %), by approximately four fold, in comparison to control (40.5 %), albeit with a slight delay (∼2 days) for obtaining fruiting bodies and a somewhat reduced total tryptamine content (0.85 %) as gauged by High Performance Liquid Chromatography measurements. Supplementation with both casing and gypsum, however, appears to promote maximal yields (896.6 g/kg of dried substrate), with a biological efficiency of 89.6 %, while also maintaining high total tryptamine expressions (0.95 %). These findings, revealing methods for maximizing yield of harvest and expressions of psychoactive tryptamines, may prove useful for both home growers and commercial cultivators of this species, and ultimately support the growth of a robust industry with high quality natural products.

Exploring the Antioxidant Potential of Methanolic Extracts of Wild Medicinal and Edible Mushrooms from Darma Valley, Pithoragaph, Kumaun (Himalaya, India).

This study presents a comprehensive analysis of the methanolic extracts of nine species of wild edible mushrooms (WEM) native to the Darma Valley in the Kumaun Himalaya region. The investigation encompasses the assessment of various biochemical attributes, including total phenolics (TP), total flavonoids (TF), total tannins (TT) contents, the ABTS assay, and the DPPH radical scavenging assay. Among the nine WEM species examined, Clavatia craniiformis stands out for displaying the highest antioxidant capacities, indicated by exceptional TP (54.94 ± 0.54 mg gallic acid equivalenta/g dry weight) and TT (4.23 ± 0.17 mg tannic acid equivalents/g dry weight) contents, along with noteworthy ABTS (10.44 ± 0.34 mg abscorbic acid equivalents/g dw) and DPPH activity (0.335 ± 0.001 mg abscorbic acid equivalents/g dry weight). Subsequent antioxidant potential are mushrooms Ramaria fennica, Ramaria botrytis, Ramaria sanguinea, Ramaria flava, Gomphus. clavatus, Clavaria zollingeri, Pleurotus ostreatus, and Kuehneromyces mutabilis. Variations in antioxidant capacities align with distinct phenolic content. This study underscores as a remarkable source of antioxidants, suggesting its potential suitability for nutraceutical applications. The findings contribute to a deeper understanding of the antioxidant properties inherent in wild edible mushrooms, particularly emphasizing the prominence of C. craniiformis.

Antioxidative and Antibacterial Hydro-Ethanolic Fraction from an Asian Edible Mushroom Lentinus sajor-caju (Agaricomycetes) Suppresses Inflammatory Responses by Downregulating COX-2 and iNOS Expression.

Mushrooms are prevalently important sources of pharmaceutically active metabolites. Various mushroom species belonging to the Lentinus genus are recognized for their nutritional and therapeutic properties. One such species is L. sajor-caju, which is renowned in Southeast Asian nations for its culinary value. The primary goal of this study is to investigate the potential medicinal properties of L. sajor-caju, specifically its antioxidant, antibacterial, and anti-inflammatory effects. A hydroethanolic extract was formulated using dried basidiocarps, which exhibited a high phenolic content of approximately 14% and a flavonoid content of approximately 2.7%. The extract demonstrated significant antioxidant potential in in vitro reactions. The extract is sufficiently capable of scavenging free radicals (DPPH and ABTS) and chelate Fe2+ with EC50 values spanning from 186 to 390 µg/mL. In addition, considerable antimicrobial activity against tested pathogenic microorganisms was observed, as indicated by low MIC50 values (256-358 µg/mL). Moreover, the fraction was found to prevent heat-induced protein denaturation which signifies its anti-inflammatory potential. When tested on the RAW 264.7 cell line, reduction in the nitrite production, and downregulation of COX-2 and iNOS mRNA expression was observed which are the key regulator of inflammatory signalling systems. The study, therefore, recommends the use of L. sajor-caju in the medical and pharmaceutical industries for the benefit of humanity.

Multiple fingerprint and pattern recognition analysis on polysaccharides of four edible mushrooms.

Quality analysis of edible mushrooms based on polysaccharides is generally difficult due to their complicated structures and hard separation. Here, multiple fingerprint analysis of polysaccharides based on chromatographic and spectrometric techniques were developed, and then applied in comparative analysis of Auricularia heimuer (AH), Auricularia cornea (AC), Auricularia cornea 'Yu Muer' (ACY) and Tremella fuciformis (TF). Firstly, polysaccharides were obtained with the molecular weights between 1.783 × 106 and 6.774 × 106 Da. Then, complete hydrolysis by TFA and enzyme digestion by cellulase were employed and subsequently analyzed by HPLC-UV, GC-MS, HILIC-HPLC-ELSD and HILIC-HPLC-ESI--HCD-MS/MS, and ATR-FT-IR were used to characterize the functional groups of intact polysaccharides. By chemometric analysis, differential markers of d-xyl, l-fuc, l-arb, d-glc, disaccharide and hexasaccharide were selected, and AC and ACY were proved to be same species from the viewpoint of polysaccharides firstly. Furthermore, the structures of oligomers with DPs of 2-8 and →4)-ß-d-Glcp-(1→ unit with different contents were inferred by combinatory analysis of ESI--MS/MS, glycosidic linkage, monosaccharide compositions and functional groups. In conclusion, the combinatory method of multiple fingerprint and pattern recognition is powerful not only for structural elucidation of polysaccharides, but also for quality analysis and species differentiation of edible mushrooms from the perspective of biological polysaccharides.

Acute hepatic and kidney injury after ingestion of Lepiota brunneoincarnata: Report of 2 cases.

Lepiota brunneoincarnata is a highly toxic mushroom species known to cause acute liver failure. However, there are limited reports investigating L. brunneoincarnata causing acute hepatic and renal damage. The present article reports 2 cases of L. brunneoincarnata poisoning in a mother and son from Chuxiong City, Yunnan Province, China. Both patients presented with gastrointestinal symptoms approximately 8-9 h after ingesting the suspect mushrooms and sought medical attention 27-28 h post-ingestion, both exhibiting acute hepatic and kidney injuries. Morphological and molecular biology studies confirmed the species of the mushrooms as L. brunneoincarnata. Liquid chromatography-tandem mass spectrometry analysis revealed mean fresh-weight concentrations of 123.5 µg/g α-amanitin and 45.7 µg/g ß-amanitin in the mushrooms. The patients underwent standard treatments, including multiple-dose activated charcoal, oral silibinin capsules, N-acetylcysteine, penicillin G, hemoperfusion, and plasma exchange. One patient recovered completely and was discharged after 16 days of hospitalization. The other patient exhibited gradual improvement in liver and renal function; however, renal function deteriorated 9 days after ingestion, and the patient declined renal replacement therapy and returned home 14 days post-ingestion. The patient was then re-hospitalized due to oliguria and edema in both lower extremities. Renal biopsy revealed acute tubular necrosis, inflammatory cell infiltration, minor glomerular capsular fibrosis, loss of microvilli in the renal tubular epithelial cells, and interstitial edema. The patient underwent 2 rounds of continuous renal replacement therapy, which eventually resulted in improvement, and was discharged 31 days after mushroom consumption. It is noteworthy that this patient had already progressed to chronic kidney insufficiency 11 months after intoxication.

Determination of psilocybin and psilocin content in multiple Psilocybe cubensis mushroom strains using liquid chromatography - tandem mass spectrometry.

A method for clinical potency determination of psilocybin and psilocin in hallucinogenic mushroom species Psilocybe cubensis was developed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Five strains of dried, intact mushrooms were obtained and analyzed: Blue Meanie, Creeper, B-Plus, Texas Yellow, and Thai Cubensis. An extraction protocol was developed; this included an evaluation of sample milling technique, extraction solvents, and recovery/stability. Reversed phase chromatography on fused-core particle phases was developed for the determination of the two analytes using internal standard calibration with deuterated isotopologues of each analyte. The separation takes less than 5 min. Matrix effects were investigated by comparing signal response of calibration samples in neat solution and several mushroom matrices; no significant matrix effects were observed. The limit of detection for psilocybin was 1.5 ng/mL (1.5 pg on-column; 300 ng/g mushroom) and for psilocin was 0.15 ng/mL (0.15 pg on-column; 30 ng/g mushroom) using a Shimadzu LCMS-8050 triple quadrupole mass spectrometer. Assessment of the accuracy and precision of the method indicated percent error and RSD were <6 % at all concentration levels. Three whole, intact mushrooms from each strain were analyzed individually to obtain average content differences both between strains and between mushrooms of the same strain. From most to least potent, the study found that the average total psilocybin and psilocin concentrations for the Creeper, Blue Meanie, B+, Texas Yellow, and Thai Cubensis strains were 1.36, 1.221, 1.134, 1.103, and 0.879 % (w/w), respectively. A subset of these mushrooms was also tested in a separate non-affiliated laboratory, and the results were comparable between the two laboratories. Results from the secondary laboratory showed improved precision when multiple mushrooms were homogenized together, prior to extraction.

Evaluation of spontaneous exploratory and anxiety-related behaviors of mice treated with gymnopilins obtained from the mushroom Gymnopilus imperialis (Agaricomycetes, Basidiomycota).

Gymnopilins are long chain oligoisoprenoids produced through the condensation of isoprene units from MEV and MEP biosynthetic pathways. In Gymnopilus, these carotenoid-like molecules are recognized as major compounds in some species. In the present study, oligoisoprenoids derived from gymnopilins were dereplicated from Gymnopilus imperialis, a mushroom-forming basidiomycete, using liquid chromatographic coupled with high-resolution mass spectrometry (tandem LC-HRMS/MS) and GNPS. From the dichloromethane extract (Gym-DCM) of G. imperialis we annotated 3 oligoisoprenoids from the GNPS molecular library spectra and 15 analogs from the curation of the molecular networking. Data from NMR spectroscopic of the extract confirmed the annotation of the metabolites. Based on the literature data suggesting the neurotoxic effect of gymnopilins, we investigated the effects of the administering different doses of gymnopilin extracts (1, 4 or 10 mg/kg) and diazepam (4 mg/kg) on the acquisition of object recognition memory (ORM) in mice. By studying novel object recognition memory (ORM), a type of non-aversive memory. ORM was assessed based on the total time of spontaneous exploration of both objects, the discrimination index (DI), and the frequency of contact with both objects. Our present findings reveal, for the first time, that gymnopilins treatment before training modulates ORM in a dose-dependent manner. It is also suggested that differential effects on memory might be related to differential effects on GABAA receptors but do not exclude its effects in other neurotransmitter systems. Another class of secondary metabolites, alkaloids, might modulate AChR, which is essential for maintaining object recognition memory over time.

An insight into the nutritional and medicinal value of edible mushrooms: A natural treasury for human health.

Edible mushrooms have been cherished worldwide because of their nutraceutical and medicinal properties. They are recognized as the new superfood for the future due to their low-calorie content, high-protein content, low lipid levels, low cholesterol levels, and abundance of essential vitamins. The fruiting body of edible mushrooms contains a plethora of primary and secondary metabolites. However, submerged cultivation is a more reliable and controlled way of production of mycelium biomass and many bioactive compounds. Several bioactive metabolites present in mushrooms possess a range of beneficial properties, including antioxidant, antimicrobial, anticancer, antidiabetic, anti-inflammatory, antiviral and anti-COVID-19 activities. Consumers have turned more intrigued in mushroom-containing products as the world needs to diversify its protein sources to meet the growing demand for protein. In this context, mushrooms are viewed as a promising source of bioactive chemicals that can be employed as an alternative to meat products. This review aims to summarise the most recent data regarding the beneficial health effects and the development of mushroom-based food products.

Antiphytopathogenic activity of the bioluminescent mushroom Neonothopanus nambi against root-rot disease.

Aurisin A (1) and the culture medium of the luminescent mushroom Neonothopanus nambi showed antifungal activity against Phytophthora palmivora, which causes the root-rot disease of Monthong durian, in a preliminary greenhouse experiment. Moreover, a new natural product, neonambiquinone B (2), was isolated. Their structures were elucidated by mass, IR and extensive analysis of their 1D and 2D NMR spectroscopic data. The results demonstrated that the culture medium of N. nambi is a promising potential for their agricultural applications.

Potential role of ergothioneine rich mushroom as anti-aging candidate through elimination of neuronal senescent cells.

Oxidative stress can upset the antioxidant balance and cause accelerated aging including neurodegenerative diseases and decline in physiological function. Therefore, an antioxidant-rich diet plays a crucial role in healthy aging. This study aimed to identify and quantify mushrooms with the highest ergothioneine content through HPLC analysis and evaluate their anti-aging potential as a natural antioxidant and antisenescence in HT22 cells. Among the 14 evaluated mushroom species, Lentinula edodes (LE), shiitake mushroom contains the highest ergothioneine content and hence was used for the in-vitro studies. The cells were preincubated with ethanolic extract of ergothioneine-rich mushroom and the equimolar concentration of EGT on t-BHP-induced senescence HT22 cells. The extract was analyzed for its free radical scavenging properties using DPPH and ABTS methods. Then, the neuroprotective effect was conducted by measuring the cell viability using MTT. Senescence-associated markers and ROS staining were also analyzed. Our results revealed that a low dose of t-BHP reduces cell viability and induces senescence in HT22 cells as determined through ß-galactosidase staining and expressions of P16INK4a, P21CIPL which are the markers of cellular senescence. However, the pretreatment with ethanolic extract of LE for 8 h significantly improved the cell viability, reversed the t-BHP-induced cellular senescence in the neuronal cells, and reduced the reactive oxygen species visualized through DCFH-DA staining. These results suggest that ergothioneine-rich mushroom is a potential candidate for anti-aging exploration through the elimination of senescent cells.

Key components, formation pathways, affecting factors, and emerging analytical strategies for edible mushrooms aroma: A review.

Aroma is one of the decisive factors affecting the quality and consumer acceptance of edible mushrooms. This review summarized the key components and formation pathways of edible mushroom aroma. It also elaborated on the affecting factors and emerging analytical strategies of edible mushroom aroma. A total of 1308 volatile organic compounds identified in edible mushrooms, 61 were key components. The formation of these compounds is closely related to fatty acid metabolism, amino acid metabolism, lentinic acid metabolism, and terpenoid metabolism. The aroma profiles of edible mushrooms were affected by genetic background, preharvest factors, and preservation methods. Molecular sensory science and omics techniques are emerging analytical strategies to reveal aroma information of edible mushrooms. This review would provide valuable data and insights for future research on edible mushroom aroma.

Quantitation of total vitamin D2 and D4 in UV-exposed mushrooms using HPLC with UV detection after novel two-step solid phase extraction.

A robust method for quantitation of total vitamin D2 and D4 in mushrooms by high performance liquid chromatography with UV detection (HPLC-UV) was developed to analyze mushrooms exposed to UV light. A two-step solid phase extraction (SPE) (silica, carbon black) removed chromatographic interferences typically resolved only with mass spectrometric detection (LC-MS) and allowed quantitation of all vitamin D and pre-D analytes. The vitamin and pre-vitamin forms of D2, D4 and D3 (internal standard), as well as other photoisomers and sterols were resolved. Results for six types of UV-exposed mushrooms were comparable to LC-MS. Screening of ten additional types of UV-exposed mushrooms without the IS confirmed lack of interference with the IS. The limit of quantification (µg/100 g fresh weight) was 0.4 for vitamin D and 0.9 for pre-vitamin D. Mushrooms do not have to be dried, and separatory funnels and large solvent volumes were also eliminated from sample preparation.