Lotion Research Articles

Next-generation risk assessment read-across case study: application of a 10-step framework to derive a safe concentration of daidzein in a body lotion.

Introduction: We performed an exposure-based Next Generation Risk Assessment case read-across study using New Approach Methodologies (NAMs) to determine the highest safe concentration of daidzein in a body lotion, based on its similarities with its structural analogue, genistein. Two assumptions were: (1) daidzein is a new chemical and its dietary intake omitted; (2) only in vitro data were used for daidzein, while in vitro and legacy in vivo data for genistein were considered. Methods: The 10-step tiered approach evaluating systemic toxicity included toxicokinetics NAMs: PBPK models and in vitro biokinetics measurements in cells used for toxicogenomics and toxicodynamic NAMs: pharmacology profiling (i.e., interaction with molecular targets), toxicogenomics and EATS assays (endocrine disruption endpoints). Whole body rat and human PBPK models were used to convert external doses of genistein to plasma concentrations and in vitro Points of Departure (PoD) to external doses. The PBPK human dermal module was refined using in vitro human skin metabolism and penetration data. Results: The most relevant endpoint for daidzein was from the ERα assay (Lowest Observed Effective Concentration was 100 ± 0.0nM), which was converted to an in vitro PoD of 33nM. After application of a safety factor of 3.3 for intra-individual variability, the safe concentration of daidzein was estimated to be 10nM. This was extrapolated to an external dose of 0.5μg/cm2 for a body lotion and face cream, equating to a concentration of 0.1%. Discussion: When in vitro PoD of 33nM for daidzein was converted to an external oral dose in rats, the value correlated with the in vivo NOAEL. This increased confidence that the rat oral PBPK model provided accurate estimates of internal and external exposure and that the in vitro PoD was relevant in the safety assessment of both chemicals. When plasma concentrations estimated from applications of 0.1% and 0.02% daidzein were used to calculate bioactivity exposure ratios, values were >1, indicating a good margin between exposure and concentrations causing adverse effects. In conclusion, this case study highlights the use of NAMs in a 10-step tiered workflow to conclude that the highest safe concentration of daidzein in a body lotion is 0.1%.

Development of a versatile optical pH sensor array for discrimination of anti-aging face creams

The certification of cosmetic products has always been a prominent concern. Here, we have developed a pH sensor and applied it in the field of cosmetic safety. Initially, we designed two probes, CH with aggregation-induced emission (AIE) effect and the near-infrared fluorophore derivative CYTYR. By encapsulating them with DSPE-PEG2000-NH2, we obtained the CHCY-lipo nano-micelles with fluorescence resonance energy transfer (FRET) response. By combining them into a sensor array called pC, we achieved sensitive detection of a wide pH range, ranging from 4.69 to 9.25. To validate the performance of the pC sensor array, we employed a multi-channel mode and applied it to differentiate commercial anti-aging creams. Through linear discriminant analysis and 3D fingerprint analysis, the pC sensor array successfully distinguished anti-aging creams from different countries, providing a rapid and accurate method for cosmetic safety identification. The results of this study demonstrate the potential of the pC sensor array for quick authentication of cosmetic products, offering significant support and application prospects in safeguarding consumer health.

Preparation and Evaluation of Aloe-Vera and Vitamin E Anti-aging cream

Aging is a natural progressive process that leads to aesthetic and functional changes in the skin. Premature aging is also observed at young age around 20 due to pollution, cigarette smoke, sunlight, effects of an unhealthy lifestyle, insufficient nutritional intake, and prolonged illness. The aim of this research work is to formulate and evaluate an herbal antiaging face cream using Aloe-Vera and Vitamin E. Radical scavenging activity of prepared cream was studied using DPPH method and standard ascorbic acid was used. The evaluation test like, Organoleptic Test, pH, viscosity, spreadability, phase separation, homogeneity test, cream type, Draize test and ease to removal test were carried out. The stability study over three week was also executed.

The Effect of Promotion Through the Shopee Marketplace on Purchasing Decisions of Scarlett Body Lotion among Students

The purpose of this study was to determine the effect of promotion on purchasing decisions. This type of research is descriptive quantitative, using a correlation approach and simple linear regression. The population in this study were students from the Faculty of Social and Political Sciences, Sam Ratulangi University Manado with sample criteria, namely students who have used or bought Scarlett Body Lotion products at Shopee, have bought similar products from the Scarlett Whitening brand, and have seen or visited advertisements about these products at Shopee. The research sample consisted of 97 respondents using a Likert scale questionnaire instrument. The results of the research in the t test show that there is a significant and positive effect of Promotion on Purchasing Decisions.

Application of Sacha Inchi Oil and Deterpenated Nutmeg Oil as Natural Antiaging in Facial Cream Formulation

Abstract Sacha inchi oil (Plukenetia volubilis L.) is rich in essential nutrients such as omega-3 fatty acids (α-linolenic acid), omega-6 (linoleic acid), omega-9 (oleic acid) and vitamin E (α-tocopherol and δ -tocopherol) which acts as a good skin barrier and reduces skin inflammation due to UV exposure. On the other hand, deterpenated nutmeg oil was reported to have antioxidant activity (IC50) stronger than native nutmeg oil. This study aims to utilize sacha inchi oil and deterpenated nutmeg oil as natural antiaging in facial cream formulations. Before used, sacha inchi oil is subjected to chemical glycerolysis to improve consumer acceptance. The physic-chemical properties between sacha inchi native oil and after pre-treatment were not significantly different. However, the hedonic test showed that the glycerolysis pre-treatment at 80°C had a better acceptance than native sacha inchi oil. The facial cream formulation used 5% sacha inchi oil and 1% deterpenated nutmeg oil with 1 – 5% niacinamide. All formulations showed oil in water (o/w) emulsion type. Facial cream with 5% niacinamide was the best formulation based on the hedonic test. It has a pH of 7.58, spreadability of 5.51 cm, and globule size of 7.11 μm

Whole-cell Lipase Catalytic Synthesis of Short-chain Fragrance Esters using Aspergillus flavus

Background: Fragrances are the collection of unlike functional assemblies, most likely alcohols, esters, aldehydes, ketones, and acids in organic products/hydrocarbons. Short-chain aliphatic fragrance esters have immense applications as flavors in the food, pharmaceutical and cosmetic industries and also have remarkable commercial significance in cosmetics and personal care products like perfumes, face creams, shampoos, soaps, lotions, jams, jellies, etc. Objective: This study aimed to synthesize short-chain fragrance esters using a whole-cell lipase catalyst from Aspergillus flavus (RBD-01). Method: The present study emphasizes the synthesis of artificial flavoring compounds by using a wholecell biocatalytic process, which can have wide significance. Herein, the preparation of ethyl alkanoates (ethyl propanoate to ethyl decanoate) was performed to investigate the flavors and fragrance excellence. The biomass from Aspergillus flavus (RBD-01) was used as a catalyst to facilitate the remarkable esterification activities towards the synthesis of important aroma esters with the help of a series of short-chain acids and alcohols. Result: The ethyl hexanoate (4) among all synthesized alkanoates was found to have a fruity fragrance with a good conversion rate. Further synthesized alkyl hexanoates (4A-4I) were found to have good fruity/pineapple/berry flavors and significant aroma quality. Conclusion: These results implied that whole-cell lipase of Aspergillus flavus (RBD-01) is a promising biocatalyst in the production of flavor aroma esters and can boost production in the food/cosmetic manufacturing industries.

Formulation of Body Lotion with Sucrose Ester Emulsifier

Abstract Sucrose ester is synthesized from palm fatty acid methyl ester and sucrose using a base catalyst. Emulsifiers that replace palm oil with sucrose esters are biodegradable and potentially non-toxic to the skin. The efficiency of sucrose ester production was obtained by using water as a solvent for sucrose, with a mass ratio of 3:5 for water and sucrose. Dilution of sucrose with water reduces caramelization during reaction. Based on laboratory research, sucrose ester has good emulsion stability from sucrose myristate ester, sucrose palmitate ester, and sucrose laurate ester. The design used a random block design with three levels of variation in sucrose ester concentration, namely 2%, 6%, and 10%. Based on ANOVA, there is a very significant effect between variations in concentration on specific gravity, pH, viscosity and surface tension. Verification tests were carried out on consumer references in the form of organoleptic tests on 35 respondents. Then, microbial tests are carried out based on Indonesian National Standards, namely the maximum number of total plates (10−2 colonies), negative tests for fungi, Staphylococcus aureus, Pseudomonas aeruginosa, and Coliform (maximum 3). Determining the best combination uses the Composite Performance Index (CPI) and Analytical Hierarchy Process (AHP). Based on organoleptic tests, the best sample is 10% myristate. The analysis of organoleptic results is consistent with the analysis of laboratory results which states that the lotion sample made from 10% sucrose myristate ester is the best sample. The best lotion samples have good emulsion stability and are not contaminated by microbes.

Formulation and Physical Quality Test of Body Lotion of Averrhoa bilimbi L. Extract as Anti-Mosquito Agent for Aedes aegypti

Formulation and Physical Quality Test of Body Lotion of Averrhoa bilimbi L. Extract as Anti-Mosquito Agent for Aedes aegypti

How Does Personal Hygiene Influence Indoor Air Quality?

Humans are known to be a continuous and potent indoor source of volatile organic compounds (VOCs). However, little is known about how personal hygiene, in terms of showering frequency, can influence these emissions and their impact on indoor air chemistry involving ozone. In this study, we characterized the VOC composition of the air in a controlled climate chamber (22.5 m3 with an air change rate at 3.2 h-1) occupied by four male volunteers on successive days under ozone-free (∼0 ppb) and ozone-present (37-40 ppb) conditions. The volunteers either showered the evening prior to the experiments or skipped showering for 24 and 48 h. Reduced shower frequency increased human emissions of gas-phase carboxylic acids, possibly originating from skin bacteria. With ozone present, increasing the number of no-shower days enhanced ozone-skin surface reactions, yielding higher levels of oxidation products. Wearing the same clothing over several days reduced the level of compounds generated from clothing-ozone reactions. When skin lotion was applied, the yield of the skin ozonolysis products decreased, while other compounds increased due to ozone reactions with lotion ingredients. These findings help determine the degree to which personal hygiene choices affect the indoor air composition and indoor air exposures.

Pharmaceutical Assessment of Body Lotion: A Herbal Formulation and its Potential Benefits

Background: Protective layers of skin cover the body. Plant-based herbal body lotion soothes and moisturises. Treatments commonly include succulent aloe vera, which heals, reduces pain, and moisturises. For hundreds of years, it has healed skin burns and injuries. Aim: This study aims on the pharmaceutical assessment of Aloe-vera by formulating an herbal Body lotion. Material and Method: Aloe-vera, Honey, Glycerin, Rose Water and Triethanolamine were taken for the formulation of herbal body lotion. Evaluation parameters were also performed to evaluate the formulation and to make sure that the subjected formulation is not harmful for the human mankind. Result: The aloe vera body lotion was formulated by using various type of ingredients such as Aloe- vera, glycerin, rose water, honey and Triethanolamine. Aloe-vera contain antimicrobial and hydrating properties protect skin against microbial degradation and moisture to skin. Conclusion: herbal body lotion is prepared for tropical administration. Aloe vera is used in lotion to provide synergistic effect as well as moisturizing effect on skin. Herbal remedies are experiencing a surge in popularity worldwide. The utilization of aloe vera, honey, Coconut oil, Lemon Oil and glycerin in the formulation of an herbal lotion is an exemplary notion. Keywords: Herbal body lotion, aloe-vera, honey, skin, glycerin, pharmaceutical assessment etc.

A Randomized Controlled Prospective Cohort Study on the Efficacy of a Witch Hazel Extract Cream for the Eyelids and Eye Contour Area and a Cleansing Face Cream in Dermatitis of the Eyelids

A Randomized Controlled Prospective Cohort Study on the Efficacy of a Witch Hazel Extract Cream for the Eyelids and Eye Contour Area and a Cleansing Face Cream in Dermatitis of the Eyelids

Cosmetic degradation: A study on body lotions with botanical extracts

Lipid oxidation is a spontaneous process in fats and, consequently, in products with significant fat content. Many cosmetics fall into this category, presenting an intrinsic relationship between the productʼs quality, oxidation capacity, and market value. This study comprehensively investigated the aging of body lotions, accentuating the role of their chemical formulations. Sensory analysis clarified that controlled temperature-induced aging significantly altered the lotions' appearance, texture, color, and, to a lesser extent, odor, hinting at the onset of rancidity. As lipid peroxidation unfolds, the emergence of secondary oxidation products like aldehydes, ketones, and alcohols becomes prominent. These secondary products indicate a potential degradation in the lotion's quality and efficacy. Quantifying the primary oxidation products was possible using methods like the quantification of Conjugated Dienes (CD) expressed in percentage and the Peroxide Index (PI) method expressed in meqO2/kg of a dry sample. On the other hand, methods such as the p-Anidisine Index (pAi) and the thiobarbituric acid reactive substances (TBARS) with malonaldehyde (MDA) expressed as malonaldehyde concentration (mg/g of dry sample) provided the quantification of secondary oxidation products. Regarding lipid oxidation, lotions containing sunflower and soybean oils emerged as notably susceptible. Conversely, samples that demonstrated minimal oxidation, were attributed to protective elements such as Aloe barbadensis and Calendula officinalis extracts. Examining into secondary oxidation, four samples displayed the highest TOTOX values, indicating extensive oxidation. A discernible correlation was observed between TOTOX values and natural extracts' presence (or lack), underscoring their protective or exacerbating roles. Certain natural ingredients, including A. barbadensis leaf juice powder and C. officinalis flower extract, were recognized as supporters against oxidation. Yet, components like PEG-40 hydrogenated castor oil can, paradoxically, seem to intensify the oxidative cascade. This research provides crucial insights into the interplay between natural extracts and lipid oxidation in body lotions. The elucidated results accentuate the necessity for judicious ingredient selection, balancing the inclusion of natural extracts to ensure product quality, longevity, and safety.

Crystallization and Solvent Evaporation Ionization Mass Spectrometry (CSEI-MS) for Rapid Detection of Drugs in Complex Matrices.

Illegal addition of drugs is common but seriously threatens public health safety. Conventional mass spectrometry methods are difficult to realize direct analysis of drugs existing in some complex matrices such as seawater or soil due to the ion suppression effect and contamination to MS parts caused by nonvolatile salts. In this work, a novel crystallization and solvent evaporation ionization mass spectrometry (CSEI-MS) method was constructed and developed to achieve rapid desalting detection. CSEI only consists of a heated plate and a nebulizer and exhibits excellent desalting performance, enabling direct analysis of six drugs dissolved in eight kinds of salt solutions (up to 200 mmol/L) and three complex salty matrices. Under optimized conditions, CSEI-MS presents high sensitivity, accuracy, linearity, and intraday and interday precision. Finally, this method is applied to the quantitative analysis of drugs in seawater, hand cream, and soil. Furthermore, the highly sensitive detection of CSEI-MS is demonstrated to remain even if the detection processes are conducted within 5 s via common commercial tools.

Herbal Skin Magic: Crafting Vanishing Cream with Nature's Best

In recent years, there has been a paradigm shift in the skincare industry towards the use of natural ingredients, driven by consumer demand for safer, eco-friendly alternatives. This research paper explores the formulation and evaluation of a novel vanishing cream enriched with botanical extracts renowned for their skincare benefits. Moringa oleifera, known for its antioxidant properties and rich nutrient profile, serves as a foundational ingredient, complemented by turmeric, celebrated for its anti-inflammatory and brightening effects. Aloe vera, prized for its soothing and hydrating properties, joins the blend alongside ginger, valued for its toning and revitalizing qualities. Lemon oil, renowned for its astringent and clarifying properties, completes the botanical ensemble. The vanishing cream formulation process involves meticulous optimization to achieve the desired texture, stability, and efficacy. Each ingredient is carefully selected and balanced to synergistically enhance the overall performance of the cream. Physicochemical analysis, encompassing parameters such as pH, viscosity, and spreadability, provides insights into the cream's quality attributes and consumer acceptability. Furthermore, the formulated vanishing cream undergoes comprehensive evaluation through in vitro and in vivo studies to assess its skincare efficacy. In vitro assays investigate the cream's ability to hydrate the skin, control sebum production, and neutralize free radicals. In vivo studies involve human volunteers to assess the cream's performance in real-world conditions, including its moisturizing effect, sebum absorption, and overall skin appearance. The results of the research demonstrate the potential of the formulated vanishing cream to address key skincare concerns, including hydration, oiliness, and overall skin health. By harnessing the power of natural ingredients, the cream offers consumers a safe, effective, and sustainable solution for their skincare needs. The findings of this research contribute to the growing body of scientific literature on natural skincare formulations and highlight the importance of botanicals in cosmetic innovation

A FORMULATION OF HAND BODY LOTIO FORMULATION OF HAND BODY LOTION EXTRACT ethanol from lily leaves (Crinum asiaticum L.) AS A SKIN MOISTURIZER

Lily leaves (Crinum asiaticum L.) have bioactivity as antioxidants and contain chemical compounds such as glycosides, flavonoids, saponins, tannins and triterpenoids. This study aims to formulate daffodil leaf extract into a hand body lotion dosage form and observe the effectiveness on volunteer skin for 4 weeks. This research was conducted experimentally. Samples were taken from daffodil leaves (Crinum asiaticum L.), then extracted by maceration with 96% ethanol and concentrated using a rotary evaporator. The ethanol extract of daffodil leaves was formulated with concentrations of 5%, 8% and 11%. The preparation evaluation test includes homogeneity, organoleptic, pH, emulsion type, irritation test, and effectiveness test of the preparation to moisturize the skin. The results showed that the ethanol extract of daffodil leaves could be used to make homogeneous grated emulsion preparations which were stable during 4 weeks of storage, pH 7.52-7.25, oil-in-water emulsion type and did not disappear. Any hand cream product that contains narcissus leaf ethanol can increase skin moisture. The composition of the most effective hand and body cream in the treatment of dry skin is a preparation with a concentration of 11% ethanol extract of lily leaves, which increases skin healing by 40.35% and is applied to the skin within 1 month.

Analysis of Hydroquinone Compounds in Whitening Body Lotion Preparations Circulating in The Community

Skin care has become an important part for today's women. Whitening body lotion is very necessary in maintaining skin when doing activities outside and indoors. Many Indonesian women desire to have white, clean and bright skin so they can appear attractive. An attractive appearance is very necessary in society. To achieve this, many women choose skin care products at affordable prices without paying attention to the dangers of the chemical content in them. One of the skin whitening ingredients that is often used in beauty products is the compound hydroquinone. The hydroquinone compound plays a role in preventing skin pigmentation. However, if the levels exceed 0.02% in the preparation, it will cause skin irritation, kidney function disorders and cancer. The type of research is a mixed type, namely qualitative and quantitative. Qualitatively, it will obtain data on the presence of hydroquinone and quantitatively in the form of hydroquinone levels. The samples were 5 whitening body lotion preparations circulating in Lubuk Pakam which were determined randomly and were coded A, B, C, D, and E. Qualitative analysis was carried out using the TLC method and quantitative analysis was carried out using the UV-spectrophotometric method. Visible. The results of qualitative analysis show that samples A, C, and D contain hydroquinone compounds which are characterized by the appearance of blackish brown spots with respective Rf values of 0.89; 0.96; and 0.89. Meanwhile, when qualitative analysis was carried out, the levels obtained for the three samples were 44.56% each; 5,604%; and 5,612%. Based on regulations issued by BPOM RI in 2022, it is stated that hydroquinone levels in beauty products must not exceed 0.02%. So it can be concluded that whitening body lotion preparations with sample codes A, C, and D are not recommended for use because the use limit for hydroquinone is classified as unsafe. It is recommended that people, especially women, be more selective in choosing and using beauty products.

Valorisation of Raspberry Seeds in Cosmetic Industry-Green Solutions.

The fruit processing industry generates large quantities of by-products well known to be rich in bioactive compounds with numerous nutritional properties and beneficial effects for human health. We developed a strategy to valorise raspberry seeds and obtain valuable ingredients with potential application in cosmetic skincare formulas. Cold press extraction technology was applied to extract oil, and the remaining defatted raspberry seed cake was treated with three proline based deep eutectic solvents (DES) to extract polyphenols. The most potent was proline/citric acid extract, with free and total ellagic acid content (52.4 mg/L and 86.4 mg/L), total phenolic content (TPC, 550.1 mg GAE/L) and radical scavenging activity (RSA, 4742.7 mmol TE/L). After the direct mixing of the extract and after encapsulation with starch as a carrier, the skincare emulsion and microemulsion were characterised by irritation potential (Zein test), transepidermal water loss (TEWL), red blood cell (RBC), and DPPH antioxidant test. The resulting preparations were of improved quality in comparison to the control hand cream, with a low skin irritation effect, lower TEWL, and higher antioxidant potential. This work complies with circular economy principles and green technology standards, and represents the efficient model on how to reuse natural resources through waste minimization.

The Influence of Viral Marketing, Brand Image and Lifestyle on Purchasing Decisions for Nivea Brand Hand and Body Lotion Products (Case Study of Students at the Faculty of Economics, University of 17 August 1945 Banyuwangi)

This research aims to determine and analyze the influence of Viral Marketing, Brand Image and Lifestyle on Purchase Decisions for Nivea brand hand and body lotion products. The research population was students from the Faculty of Economics, University of 17 August 1945 Banyuwangi, with a research sample of 65 respondents. With the data obtained using the Probability Sampling technique with a simple random sampling technique and carrying out analysis which includes instrument testing, classical assumption testing, multiple linear regression testing, hypothesis testing and coefficient of determination testing. Based on the results of the analysis, conclusions can be drawn from the results of the hypothesis test F test, the Viral Marketing, Brand Image and Lifestyle variables are stated to be good and have a significant effect on the Purchase Decision variable for Nivea brand hand and body lotion products. The results obtained include validity and reliability, all of which are suitable for use and can be used in further tests. The regression results obtained Y = 4.004 + 0.364X1 + 0.366X2 + 0.334X3, the simultaneous test obtained calculated F> F table, t test for all variables calculated t value> t table, the value of the Adjusted R Square coefficient of determination was 40.8%, Assumption test The classic includes the normality test for all normal variables, the multicollinearity test without all multicollinearity, the total heteroscedasticity test without heteroscedasticity.

Detection of Mercury in Skin Lightening Cosmetics Marketed in Katsina Metropolis, Katsina State, Nigeria

Deliberate use of metals as active cosmetic ingredients has been banned in many countries due to their persistent nature, toxicity and transdermal exposure over a certain period of time. Fifteen (15) samples were purposively sampled to cover the most frequently used brands as the samples for the study. The samples cover five (5) of each of the skin lightening creams, lotions and soaps from different cosmetics shops in Katsina central market and other outlets within the metropolis of Katsina. Qualitative inorganic analysis was conducted for the detection of the presence of mercury in the samples. Also quantitative analysis of the mercury was carried out using Atomic Absorption Spectrometry (AAS). The results for the qualitative inorganic analysis showed that all the samples were mercury (II) positive with 0.5N KI reagent through the formation of reddish orange precipitate, and no positive reaction of mercury was detected using 2.0N NaOH reagent. Quantitative analysis of the mercury revealed the presence of mercury in all the analyzed samples. The concentration of mercury ranged from 27.7 ± 0.0006 to 61.5 ±0. 0007 ppm. From the results obtained, it is thus evident that the analyzed samples were mercury (II) positive and above the recommended WHO permissible limit. Therefore, there may be a serious health hazard to the users of such products in Katsina metropolis. It is recommended that periodic analysis of such skin lightening products should be conducted by the regulatory bodies to ascertain their safety to the users.

Development of a method to evaluate the effects of external environments on drug stability in nails using micro-segmental analysis.

Nails can be used as an alternative to hair for examining past drug use. However, daily hand-and-nail care can eliminate the internal drugs. Therefore, we developed an evaluation method to examine the effects of the external environment on drug stability in nails using micro-segmental analysis. First, reference nails containing drugs were prepared by collecting fingernails from participants who had consumed hay-fever medicines continuously for 4months. Next, the entire free edge of a reference nail was cut into halves at the centerline; one side was stored as an untreated block, and the other was treated with various hand/nail care products. Both nail blocks were washed and segmented at 0.5-mm intervals in the width direction. Each segment in the extraction solution was crushed with stainless-steel beads, sonicated, and soaked in the solution for 24 h. The analytes in extracts were quantified by LC-MS/MS, and the drug concentrations between the treated and untreated blocks were compared. The drug concentrations decreased slightly in nails treated with manicure and gel-nail products. The analytes in nails tended to be lower in water-rich products such as hand soap and hand cream than in oil-rich products such as nailcare oil and acetone-free remover. The developed method using micro-segmental analysis enabled the evaluation of the effects of various hand/nail care products on drug stability in a limited number of nails. This would also be useful for examining the effects of severe environments on drugs in nails collected from cases of unnatural death.