Development and Validation of an HPLC Method for Simultaneous Determination of Capsaicinoids and Camphor in Over-the-Counter Medication for Topical Use.

A reverse-phase high-performance liquid chromatography method was developed to determine and quantify capsaicin (trans-8-methyl-N-vanillyl-6- nonenamid), dihydrocapsaicin (8-methyl-N-vanillylnonanamide), and camphor (trimethylbicyclo[2.2.1]heptan-2-one). It is applicable in analyses of over-the-counter (OTC) medications for topical use and raw materials such as chili pepper oleoresin. Chromatographic separation was carried out on a C18 column using an isocratic flow of the mobile phase containing acetonitrile and ultrapure water in a ratio of 2:3, with pH adjusted to 3.2 using glacial acetic acid, and a flow rate of 1.5 mL/min. The concentration of the eluting compounds was monitored by a diode-array detector at a wavelength of 281 nm. The method was evaluated for several validation parameters, including selectivity, accuracy (confidence intervals < 0.05%), repeatability, and intermediate precision. The limit of detection (LOD) was determined to be 0.070 µg/mL for capsaicin, 0.211 µg/mL for dihydrocapsaicin, and 0.060 µg/mL for camphor. The limit of quantification (LOQ) was determined to be 0.212 µg/mL for capsaicin, 0.640 µg/mL for dihydrocapsaicin, and 0.320 µg/mL for camphor. Linearity was set in the range of 2.5-200 µg/mL for capsaicin and dihydrocapsaicin and 25-2000 µg/mL for camphor. The suggested analytical method can be used for quality control of formulated pharmaceutical products containing capsaicinoids, camphor, and propolis.

Phototoxic risk assessment of dermally-applied chemicals with structural variety based on photoreactivity and skin deposition.

Combined use of photochemical and pharmacokinetic (PK) data for phototoxic risk assessment was previously proposed, and the system provided reliable phototoxic risk predictions of chemicals in same chemical series. This study aimed to verify the feasibility of the screening system for phototoxic risk assessment on dermally-applied chemicals with wide structural diversity, as a first attempt. Photochemical properties of test chemicals, 2-acetonaphthalene, 4'-methylbenzylidene camphor, 6-methylcoumarin, methyl N-methylanthranilate, and sulisobenzone, were evaluated in terms of UV absorption and reactive oxygen species (ROS) generation, and PK profiles of the test chemicals in rat skin were characterized after dermal co-application. All test chemicals showed strong UVA/B absorption with molar extinction coefficients of over 3000 M-1⋅cm-1, and irradiated 2-acetonaphthalene, 6-methylcoumarin, and methyl N-methylanthranilate exhibited significant ROS generation. Dermally-applied 2-acetonaphthalene and 4'-methylbenzylidene camphor indicated high and long-lasting skin deposition compared with the other test chemicals. Based on the photochemical and PK data, 2-acetonaphthalene was predicted to have potent phototoxic risk. The predicted phototoxic risk of the test chemicals by integration of obtained data was mostly consistent with their in vivo phototoxicity observed in rat skin. The screening strategy employing photochemical and PK data would have high prediction capacity and wide applicability for photosafety evaluation of chemicals.

Camphor white oil induces tumor regression through cytotoxic T cell-dependent mechanisms.

Bioactive derivatives from the camphor laurel tree, Cinnamomum camphora, are posited to exhibit chemopreventive properties but the efficacy and mechanism of these natural products are not fully understood. We tested an essential-oil derivative, camphor white oil (CWO), for anti-tumor activity in a mouse model of keratinocyte-derived skin cancer. Daily topical treatment with CWO induced dramatic regression of pre-malignant skin tumors and a two-fold reduction in cutaneous squamous cell carcinomas. We next investigated underlying cellular and molecular mechanisms. In cultured keratinocytes, CWO stimulated calcium signaling, resulting in calcineurin-dependent activation of nuclear factor of activated T cells (NFAT). In vivo, CWO induced transcriptional changes in immune-related genes identified by RNA-sequencing, resulting in cytotoxic T cell-dependent tumor regression. Finally, we identified chemical constituents of CWO that recapitulated effects of the admixture. Together, these studies identify T cell-mediated tumor regression as a mechanism through which a plant-derived essential oil diminishes established tumor burden.

Blood Pressure and Cognitive Performance After a Single Administration of a Camphor-Crataegus Combination in Adolescents with Low Blood Pressure.

Low blood pressure is rather widespread among adolescents and frequently accompanied by complaints. Two single-center, double-blinded, randomized, and placebo-controlled trials were performed with adolescent participants in the age range of 14 - 17 y having a systolic blood pressure below 118 mmHg (boys) or 110 mmHg (girls). They received a single dose of 20 drops of a fixed combination of natural D-camphor and an ethanolic extract from fresh Crataegus berries (CCC) or of an ethanolic placebo. The efficacy of CCC (commercial name Korodin) has proven its efficacy in the treatment of low blood pressure in adults of all ages, although related placebo-controlled, double-blind studies for adolescents following European (EU) pediatric regulations are lacking. Blood pressure and heart rate were assessed during a rest period, prior to substance administration, 1 min thereafter, and after about 5 min. Additionally, performance was assessed by two cognitive tests. After administration of CCC, a significantly greater rise in blood pressure occurred compared to placebo. In the cognitive tasks, no significant differences were observed. No adverse events or subjective complaints were reported at the final examination; thus, the present study provides evidence for the safety and tolerability of CCC after a single administration.

[Raspail, Don Quixote of camphor!]. / Raspail, Don Quichotte du camphre!

François Vincent Raspail (1794-1878) was a chemist and a politician who practised medicine illegally : for him, all pathologies were almost exclusively caused by "parasites" and he treated them all thanks to the beneficial effects of camphor. Raspail did not invent nor discover that substance, which was broadly used in the 18th century, but he codified the use of its various pharmaceutical forms thanks to what would be later called "Raspail's method": camphor to eat (lumps), to snuff (powder), to smoke (cigarettes), brandy and 32° camphored alcohol, camphored oil and sedative water... Since Raspail lavished his patients with many hygiene pieces of advice and moral principles, this method--that was actually a prolongation of his political action in favour of the very poor--was a complete success among the population. But this success was also due to the simplicity of the remedy, its relative harmlessness and, above all, to an incredibly efficient advertising since he enjoyed the support of many important people and also published a very successful book, his Manuel annuaire de Santé, that was re-edited 77 times.

Camphor induces cold and warm sensations with increases in skin and muscle blood flow in human.

Application of camphor to the skin has been empirically thought to improve blood circulation. However, camphor's effects on blood circulation to the skin and on thermal sensation have not been well elucidated. In this study, we examined its effects on the quality of sensation as well as on skin and muscle blood flow in human. Nine adults (average age 37±9.4 years) participated in the study. Petroleum jelly containing 5%, 10%, 20% camphor, or 2% menthol was separately applied to the skin on the medial side of one forearm of each subject. Just after the application, camphor at each concentration induced a cold sensation in a dose-dependent manner. Within 10 min, each subject reported that the cold sensation had faded, after which it was replaced by a warm sensation. As reported previously, a cold sensation was induced by application of 2% menthol, but the subjects did not adapt to that sensation. In addition, menthol did not induce a warm sensation at all. Application of menthol has been shown to increase blood flow in the skin. Finally, we measured blood flow in skin and muscle after the application of camphor or menthol. Application of camphor or menthol separately induced increases in local blood flow in the skin and muscle. The present results indicate that camphor induces both cold and warm sensations and improves blood circulation.

Randomized placebo controlled trial of phytoterpenes in DMSO for the treatment of plantar fasciitis.

Plantar fasciitis is the most common cause of heel pain in adults with an overall prevalence of 0.85% in the adult population of the US, affecting over 2 million adults annually. Most current treatment modalities are not supported by sufficient evidence to recommend one particular strategy over another. Topical application of analgesics for soft tissue pain is well established, however the plantar fascia presents challenges in this regard due to thick skin, fibrotic tissue, and an often thickened fat pad. Sixty-two patients with plantar fasciitis were randomized to a placebo controlled trial testing the efficacy of a topical solution of plant terpenes containing camphor, menthol, eugenol, eucalyptol, and vanillin. Skin permeation of the mixture was enhanced with 15% dimethylsulfoxide (DMSO), 1% limonene, and rosemary oil. One ml of solution was applied topically twice daily, and pain scores evaluated on Day 0, Day 1, Day 3, and Day 10. Using the validated foot function index 78.1% of patients reported an 85% or greater decrease in their total pain score by day 10 while placebo treatment was without effect (One Way ANOVA, P < 0.01). This study adapts the treatment modality of topical analgesia for soft tissue pain to a problematic area of the body and shows therapeutic promise.ClinicalTrials.gov Identifier: NCT05467631.

Estrogenic activity of ternary UV filter mixtures in fish (Pimephales promelas) - an analysis with nonlinear isobolograms.

Numerous estrogenic compounds are present in aquatic environments, but currently it is not well understood how compounds that differ in maxima and slope of their individual dose-response curves contribute to the overall mixture effect. In order to better understand such interactions we investigated 3 commonly used UV filters, for their estrogenic mixture activity and analysed their joint effects by using the concentration addition (CA) concept. Thereby, we extended the method of isoboles for analysis of 3 compounds that differ in maxima and slopes of their dose-response curves. 3-Benzylidene camphor (3BC), benzophenone-1 (BP1) and benzophenone-2 (BP2) are estrogenic in fish and act as pure- or partial estrogen receptor alpha agonists. First we exposed juvenile fathead minnows for 14 days to six concentrations of each UV filter alone to determine vitellogenin (VTG) induction curves, calculate equi-effective mixture concentrations and predict mixture effects. For 3BC, BP1 and BP2 significant VTG-induction occurred at 420, 2668, and 4715 microg/L, respectively. BP2 displayed a full dose-response curve, whereas 3BC and BP1 showed submaximal activity of 70 and 78%, respectively. Second, we exposed fish to 6 equi-effective mixtures (EC-NOEC, EC1, EC5, EC10, EC20, EC30) of these UV filters. Significant VTG-induction occurred at EC5 and higher. Submaximal activity of 67% as compared to the control EE2 (100 ng/L) was reached. The curves for the observed and predicted mixture activity agreed for mixture levels (EC10 to EC30), however, at EC-NOEC, EC1 and EC5, lower activity was observed than predicted by CA. Detailed isobolographic analysis indicate additivity at EC10 to EC30, and antagonism at low levels (EC-NOEC to EC5). Our data show for the first time, that for compounds with differences in maxima and slope, considerably more mixture combinations are additive than previously thought. This should be taken into account for hazard and risk assessment of UV filters and xenoestrogens.

Effects of transient receptor potential (TRP) channel agonists and antagonists on slowly adapting type II mechanoreceptors in the rat sinus hair follicle.

The possible functional role of transient receptor potential (TRP) channels was investigated by testing various TRP agonists and antagonists in an isolated rat sinus hair follicle preparation. Extracellular recordings from slowly adapting type II mechanoreceptor units were made. The antagonist capsazepine depressed spontaneous and mechanically evoked activity, with an IC(50) of 82 microM. In one-third of units, capsazepine caused a selective depression of mechanically evoked firing, such that the existing spontaneous firing was interrupted by an absence of activity during the mechanical stimulus. The broad spectrum TRP blocker ruthenium red (30 microM) had inconsistent effects, although in some units a delayed onset (following wash) bursting and paroxysmal firing ensued. The agonist icilin (50-100 microM) had an excitatory effect on spontaneous firing, and (-)-menthol (200 microM) had inconsistent effects. Cinnamaldehyde (1-2 mM) depressed all types of activity equally, mechanically evoked and spontaneous. Camphor (0.5-2 mM) also depressed all types of activity, although it had a preferential effect on spontaneous activity. Capsaicin (1-10 microM) and allyl isothiocyanate (50-100 microM) had no clear effects. These results rule out any role for TRPA1 and TRPV1 channels in mechanotransduction processes of slowly adapting type II mechanoreceptors.

Skin disposition of d-camphor and l-menthol alone and together.

The terpenes camphor and menthol are often used in topical preparations, although some data indicate concern about their skin penetration after application in most commonly used vehicles. The cutaneous disposition of these substances applied alone and together in either an oily solution or a hydrogel was evaluated ex vivo using full human skin mounted in flow-through diffusion cells. After 0.5, 1 and 2 h of application, the skin was progressively tape-stripped into three fractions of stratum corneum (SC) and the remaining epidermis with the dermis. The content of terpenes in the skin layers was determined using gas chromatography. Different penetration into the skin layers was observed depending on the type of vehicle. The highest SC absorption was noted when terpenes were applied in hydrogel, where the total content in the SC was 200 microg/cm2 for camphor and 400 microg/cm2 for menthol, and the total skin absorption was 310 and 460 microg/cm2, respectively. The SC penetration of both terpenes from the oily solution was the same (approximately equal to 35 microg/cm2). When both terpenes were present in the hydrogel the SC absorption decreased, the amounts of camphor and menthol in the SC being 50 and 190 microg/cm2, respectively (total skin accumulation was 120 and 220 microg/cm2, respectively). Such an effect was not observed for the oily solution.

László Meduna's pilot studies with camphor inductions of seizures: the first 11 patients.

In his autobiography, László Meduna described the first session of convulsive therapy using intramuscular camphor as occurring on January 23, 1934 at Royal National Hungarian Institute of Psychiatric and Neurology at Budapest-Lipótmezo in Hungary. Unearthed records of the patients treated at this institution reveal that Meduna's dose-finding experiments began on January 2, 1934. The symptomatology and history of illness, diagnosis, socio-demographic data, the seizure characteristics, and immediate and long term outcomes of the first 11 patients are described. These first trials elicited seizures in less than half the injections. Seizures of various durations (including missed seizures) and double (tardive) seizures were recorded. Mutism, refusal to eat requiring tube feeding, and other signs of catatonia dominated the psychopathology of 7 of the first 11 patients. Two improved sufficiently to be discharged from the hospital and third patient became fit for occupational therapy. These records exhibit the meticulous systematic nature of the first human trials with induced seizures and the fortuitous nature of the first human trials with induced seizures and the fortuitous nature in patient selection of catatonic patients--an illness that is most responsive to induced seizures.

Sakel versus Meduna: different strokes, different styles of scientific discovery.

It is ironic that Laszlo Meduna and Manfred Sakel made epochal discoveries in psychiatry only two years apart because in most other ways, the two men could hardly be more different. The story of their differences and similarities invites us to consider the personal preconditions for scientific discovery. What common denominators led them to develop treatments, which represent the introduction of convulsive therapy, which remains today the most powerful treatment in psychiatry? Despite the marked differences in their personalities, Sakel and Meduna shared intellectual quickness, drive, and a willingness to take risks, three qualities that, in the case of these individuals at least, came together to revolutionize the treatment of serious psychiatric illness.

Fabrication and design of transdermal fluconazole spray.

The objective of the present study was to formulate fluconazole transdermal spray for obtaining modified drug transport using eutectic mixture, ethyl cellulose, polyethylene oxide and alcohol. The formulated products were characterized. The selection of the optimized batch was done considering the results of drug transport in the first hour, the time required for 90% drug transport, viscosity and spray angle of the formulations. The inclusion of eutectic mixture, consisting of equal parts of camphor and menthol, showed improved drug transport through rat skin. The optimized batch exhibited larger mean zone of inhibition (antifungal activity), efficient in vivo activity and short term stability.

[Traditional camphor misuse: a forgotten danger to children (about 2 cases)]. / Mésusage traditionnel du camphre: un danger oublié pour les enfants (à propos de 2 cas).

In our country, the use of traditional medicinal recipes and artisan cosmetic products is very frequent due to the high rate of illiteracy, low purchasing power and of the large number of herbalists. Camphor is a low-cost product, easily accessible and omnipresent in almost every home, making it potentially toxic in case of misuse, in particular among children. We here report two cases of intoxication induced by beauty recipe made with powdered camphor. Patients' medical history gave informations about poisoning in 2 children caused by synthetic powder made with camphor imported from China. Patient 1: little girl aged 2 months, with no previous medical history, admitted to the Emergency Department due to constant crying and refusal to eat. Clinical examination showed no abnormalities. Standard laboratory tests were normal. Neurological, digestive and cutaneous monitoring were performed. Patient 2: girl aged 6 years admitted with atonic seizure associated with syncope and foaming followed by abdominal pain and vomiting after ingesting milk. The evolution was favorable 48 hours after symptom management. Mothers reported two neighbors had received a traditional hair care recipe by a third neighbor. After that they had mixed powdered camphor with olive oil, then they had applied it to the hair of their children for one hour, thus causing the occurrence of these symptoms.

Safety Threshold Considerations for Sunscreen Systemic Exposure: A Simulation Study.

Sunscreens are regulated as over-the-counter drugs in the United States. Some sunscreen ingredients are absorbed into the systemic circulation, which raises concerns about the safety of these drugs. There is limited information on the systemic exposure for most sunscreen ingredients. This report estimates the systemic absorption of two sunscreen active ingredients, oxybenzone and enzacamene, by developing a pharmacokinetic model from published sunscreen absorption data and compares the results with safety thresholds proposed by the US Food and Drug Administration and in the literature. Our analysis indicates that systemic absorption can be substantial, and evaluation of the systemic exposure of sunscreen ingredients is warranted to better assess any long-term risks of use.

Hydroxyethyl starch-induced pruritus relieved by a combination of menthol and camphor.

Hydroxyethyl starch is a key component of many colloid volume expanders used in hypovolemic shock and otologic disease. Pruritus is a common side effect. Histopathology reveals multiple cytoplasmic vacuoles in dermal macrophages, endothelial cells, and perineural cells with electron-dense foreign material within the said vacuoles. Although classically refractory to treatment with corticosteroids and antihistamines, some benefit has been achieved with capsaicin, ultraviolet light therapy, and oral naltrexone. We present a case responsive to menthol and camphor and discuss the possible therapeutic mechanism.

Camphor ingestion: an unusual cause of seizure.

Camphor is a pleasant smelling cyclic ketone with propensity of causing neurologic side-effects especially seizures. We report two patients who after inadvertent consumption of camphor experienced an episode of generalized tonic clonic seizure. These cases highlight the importance of enquiring any intake of material (medicinal or otherwise) in every patient presenting with seizure.

[The birth of convulsive therapy]. / A görcskezelés születése.

The history of the first convulsive treatment is summarized here in commemoration of its 75th anniversary. The neuropathological and clinical findings underlying the theoretical basis of the method are reviewed, together with the case histories of the first batch of patients who underwent convulsive therapy. The early indications and effectiveness of convulsive therapy are also discussed. Finally, in a broader context, the role of convulsive treatment in the development of modern biological psychiatry and Laszlo Meduna's contribution to this development touched upon.

[The birth of biological therapy in Hungary: the story of László Meduna's first patient receiving convulsive therapy]. / Biológiai terápia születése Magyarországon: Meduna László "elsô" görcskezelt betegének története.

Convulsive treatment was introduced to psychiatry by László Meduna, a Hungarian neuropsychiatrist. In his autobiography, Meduna gave a detailed description of his first patient who underwent convulsive therapy. According to Meduna's recollections, this patient was L. Zoltán, who after 4 years of fluctuating catatonic stupor received several sessions of camphor-induced convulsive therapy resulting in full remission and discharge from the institution. In this communication, the authors reconstruct L Zoltán's case history from the original case notes, which were recovered from the Archives of the National Institute of Psychiatry and Neurology, Budapest. The case notes show that L. Zoltán received courses of convulsive treatment between 1934 and 1937, first induced with camphor and then with cardiazol. After the first course of treatment the catatonic stupor was resolved and the psychotic symptoms subsided. However, this incomplete remission lasted only for a few months and was followed by a relapse. Despite further courses of convulsive therapy, L. Zoltán never became symptom free again, could never be discharged and died in the institute in 1945. The authors attempt to explain possible reasons for the discrepancies found between Meduna's account and L. Zoltán's case notes.

Untargeted search and identification of metabolites of antiviral agent camphecene in rat urine by liquid chromatography and mass spectrometry and studying their distribution in organs following peroral administration of the compound.

Major metabolites of camphecene, a new effective antiviral agent, formed after its oral administration to rats and excreted in the urine, were found and identified using liquid chromatography coupled to mass spectrometry as well as multivariate analysis of HPLC-MS data. The metabolites were found to be camphecene glucuronide, camphecene sulfate and the corresponding iminoacid. A study of the dynamics of accumulation of camphecene and its metabolites in the liver, kidneys, lungs and brain of animals was performed. Maximum concentration of camphecene in blood and organs was reached after 1.5-2 h of its administration, and the maximal content of the agent in the organs investigated was observed in the kidneys. The content of the substance in the lungs was comparable to that in the liver. Also, camphecene was found in brain in high concentration, thus allowing assumption of its ability to penetrate the blood-brain barrier and to exert its antiviral properties in the organ. Camphecene glucuronide and iminoacid had concentration-time profiles similar to that of their precursor, their content being maximal in kidney and liver and 2-3 orders of magnitude higher than in lungs and brain. The content of camphecene sulfate was of similar level in all organs studied. The results obtained made it possible to develop recommendations for therapy with the use of camphecene.