Browsing by Author "Khumalo, Nonhlanhla"

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    Open Access
    An investigation of the association between structural proteins and lipids with scalp hair curvature
    (2025) Mukonyora, Michelle; Adeola, Henry; Khumalo, Nonhlanhla
    Scalp hair is increasingly being used as a non-invasive analytical sample in diagnostic and forensic medicine. However, variable sensitivities for detection of hair biomarkers were observed, whereby African hair generally incorporated higher drug levels than Asian and Caucasian hair types. It was hypothesized that higher absolute lipid amounts in African curly hair may affect the rate at which lipid-soluble metabolites are passively incorporated into the growing hair shaft. Furthermore, the interactions between structural proteins and lipids in scalp hair may be associated with hair curvature, though the exact mechanism is unknown. This study is the first to investigate the hair proteome and lipidome in the same group of hair samples using a multiomics approach to determine how they both relate to each other, and to hair curvature. After ethical approval, hair of varying curvature was collected from 90 participants and classified from straight to curly objectively (type I – VI) using a validated tool. Hair proteins were extracted and analysed using an ultra-high performance liquid chromatography quadrupole time of flight (UPLC-QToF) mass spectrometer and the data analysed using various bioinformatics tools and gene ontology analyses. Hair lipids were extracted and analysed using a gas chromatography-triple quadrupole (GC-QqQ) mass spectrometer, various bioinformatics tools, and lipid pathway enrichment analyses. The combined proteomics and lipidomics quantification data were analysed using both supervised and unsupervised classification methods to build hair proteolipidomic models. We identified a total of 420 hair proteins, which is comparable to the highest numbers in published hair studies. We also identified the largest number of chaperones compared to any published hair study, where nine out 15 are described for the first time in hair. Importantly, seven proteins, belonging to four protein groups (transketolase, iron-sulfur cluster scaffold homolog, proline-rich protein 29, heat shock protein family A) were differentially expressed (p-value < 0.05) and described in hair for the first time. Proline-rich protein 29 was the only differentially expressed protein with a linear increase from hair type I to VI. This is also the first study to report differentially expressed proteins between all six geometric hair types. The differentially expressed hair proteins regulate various aspects of energy metabolism, and they are involved in processes known to drive keratinocyte differentiation and maintain homeostasis in the hair follicle and skin. A total of 77,060 lipid features (RT and m/z) were found, and 3,084 were identified at the lipid species level (RT, m/z, and lipid subclass), which is the largest number of hair lipids identified in any published hair study to date. For the first time, 129 differentially expressed hair lipids (p-value < 0.01) in hair of different curvature were identified. A subset of 19 differentially expressed hair lipids belonging to nine lipid subclasses (sterol lipids, steroids, hydrocarbons, fatty acids, fatty acyl carnitines, ceramides, endocannabinoids, wax esters, and ethyl esters) were characterised and pre-validated using multiple reaction monitoring (MRM) in separate hold-out hair samples. These lipids regulate necroptosis, as well as sphingolipid signalling and metabolism (FDR < 0.05). We built hair proteolipidomic models for the first time, which revealed that the hair proteolipidome comprises several heterogeneous subclusters. Also, that proteins showed a poor correlation with hair curvature as confirmed by poor/moderate ROC scores (0.6 to 0.8), whereas lipids correlated well with hair curvature as confirmed by excellent ROC scores (0.96 to 1). The hair proteolipidome further revealed protein folding, formation of the extracellular matrix, and the Wnt/β-catenin signalling pathway as being associated with hair curvature (FDR < 0.05). In conclusion, our investigation of baseline lipid and protein profiles across various geometric hair types confirms previous data of low correlation between proteins and hair curvature. However, this study identified, for the first time, 9 novel chaperones in hair. Proline-rich protein 29, also novel, was the only protein which increased from hair type I to VI. This study also reports the largest number of lipids in any hair study, is the first to report lipid sub-classes and multi-omics which confirmed the high correlation between lipids and hair curvature. A limitation of this study is that a more robust computational quantification method for hair curvature variations was not used because it was published after this study commenced. Future studies are needed to elucidate the potential contribution of identified molecules to hair curvature variation.
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    Open Access
    Bioenergetic Evaluation of Site-specific Keloid and FKN Fibroblasts
    (2019) Chalwa, Temwani; Khumalo, Nonhlanhla; Bayat, Ardeshir; Lebeko, Maribanyana
    Excessive scarring due to fibrosis from aberrant wound healing can lead to conditions such as Keloids or Folliculitis Keloidalis Nuchae (FKN). These fibroproliferative growths pose therapeutic challenges due to their complex aetiology that has been linked to multiple genetic and environmental factors, with frequent reoccurrence following therapy. Owing to reports on an increase in ATP and Fibroblast Activation Protein-1α production in keloids, the aim of this study was to investigate whether the disease phenotypes were linked to bioenergetic changes at a cellular level in these two conditions. Patient-derived tissue biopsies were used for fibroblast cell culture models, in which cell analysis was carried out to assess phenotype and different parameters of bioenergetic cellular behaviour. In addition to FKN and the intra- and peri-lesional Keloid patient fibroblasts, normal skin and non-hypertrophic (normo-trophic) scar fibroblasts were used as negative controls. The results show statistically significant and variable growth dynamics with increased proliferation and migration in keloid fibroblasts, while FKN fibroblasts showed a statistically significant increase in proliferation but had a similar migration profile to controls. The results further show that there is a statistically significant metabolic switch towards aerobic glycolysis in the fibroblasts from the disease conditions. During functional measurement of mitochondrial parameters, an increase in oxidative phosphorylation was exhibited in the disease conditions indicating their mitochondria were still functional. An increase in basal glycolysis with a concomitant increase in the cellular maximum glycolytic capacity was also demonstrated. Furthermore, protein analysis showed an upregulation in the expression of Fibroblast Activation Protein-1α in fibroblasts from both disease conditions This study begins to give novel insight into the bioenergetics of normal scars and scarring conditions such as FKN and, adds to the knowledge on the heterogeneity of fibroblasts derived from specific lesional sites within Keloids. These findings suggest that Keloids and FKN have a switch to a metabolic phenotype of aerobic glycolysis. This increase in glycolytic flux potentially proposes glycolytic inhibitors as a mechanistic basis for the treatment of these conditions.
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    Open Access
    Blood and virus detection on barber hair clippers
    (2019) Spengane, Zandile Namhla Elizabeth; Khumalo, Nonhlanhla; Ngwanya, Mzudumile R
    Background: Bleeding from the popular clean-shave ‘chiskop’ haircut was recently reported as prevalent in South Africa (SA), a country with 6.9 million HIV-infected people. Objectives. To investigate the prevalence of barber hair clipper contamination with blood and HIV and hepatitis B viruses. Methods: Fifty barbers from three townships in Cape Town, SA, were invited to participate. One clipper from each barber was collected immediately after it had been used for a cleanshave haircut. Each clipper was rinsed with phosphate-buffered saline and then submerged in viral medium. The polymerase chain reaction (PCR) was used to identify the bloodspecific RNA marker haemoglobin beta (HBB), hepatitis B virus (HBV) and HIV. Results: The clean-shave haircut was the most common haircut requested by clients (78%). Of the clippers collected, 42% were positive for HBB, confirming detection of blood, none were positive for HIV, and 4 (8%) were positive for HBV. Two clippers (clippers 16 and 20) were positive on qualitative HBV PCR. HBV DNA from clipper 16 clustered with genotype A sequences from SA, India, Brazil and Martinique, while clipper 20 clustered with SA genotype D sequences. The clipper 20 sequence was identical to a subtype D sequence (GenBank accession AY233291) from Gauteng, SA. Conclusion: This study confirms that there is significant contamination of barber hair clippers with blood and blood-borne viruses. Hepatitis B was detected with enough DNA copies to pose a risk of transmitting infection. Although HIV was not detected in this small study, the risk of transmission should be quantified. Further studies to investigate barber clipper sterilization practices and whether the clean-shave hairstyle is an independent risk factor for HIV, HBV and hepatitis C virus infections are warranted. Public education on individual clipper ownership (as is the case with a toothbrush) should be advocated for clean-shave and blade-fade haircuts.
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    Open Access
    Determination of illegal and toxic ingredients in face creams
    (2025) Mehlala, Nandipha; Oputu Utieyin, Ogheneochuko; Khumalo, Nonhlanhla
    The increasing prevalence of counterfeit and substandard face creams on the market poses significant risks to consumer health, with some products containing illegal and toxic ingredients that can lead to serious dermatological and systemic effects. This study focuses on the identification and quantification of illegal and toxic ingredients in fifty-eight skin cosmetic creams marketed to reduce pigmentation. Samples that were selected were those labelled as ‘'Skin lightening'', Skin brightening'', Skin whitening,” or “even tone” creams, purchased from local vendors within the Cape Town Metropolis, and from an online store. This study employed advanced analytical techniques to assess the potential health risks of face creams. The High-Pressure Liquid Chromatography (HPLC) method was used to determine thirteen compounds of interest such as hydroquinone, kojic acid (KA), and steroids. Secondly, Inductively Coupled plasma mass spectrometry (ICP-MS) assisted with MARS 6 digester was employed to determine metals (mercury, lead, chromium, nickel, cadmium, arsenic, etc.). Lastly, the study assessed potential health risk of the actives and metals through predictive risk assessment models developed by United States Environmental Protection Agency (USEPA). The health risk models consider active concentration (at 95% confidence level), and the unique properties of active or metal. The United States Environmental Protection Agency calculates carcinogenic cancer risk of metals using the Cancer Slope Factor (CSF) and the Average Daily Dose (ADD). The ADD is determined based on the concentration of the metal, intake rate, exposure frequency and duration, body weight, and averaging time. The cancer risk is then calculated by multiplying the ADD by the CSF, which represents the probability of developing cancer per unit of exposure. Risks are typically expressed as a probability (e.g., 1 in 10,000) and are compared to acceptable thresholds, usually ranging from 10−6 to 10−4. This assessment accounts for the metal's carcinogenic properties, exposure pathways and population sensitivities to evaluate potential health impacts. The analysis of face cream samples revealed the presence of eleven active compounds, including several illegal and toxic substances. Steroids were the most prevalent, detected in approximately 45% of the samples, with Clobetasol propionate (CP) being the predominant steroid, accounting for 68% of the steroid detections. Other steroids identified included Cortisone, Betamethasone Dipropionate (BD), Betamethasone (BM), and Fluocinolone, found in 12%, 12%, 4%, and 4% of the samples, respectively. Hydroquinone (HQ) was present in 16% of the samples, with concentrations ranging from 0.096% to 4.301%, particularly high in imported products. Arbutin, a derivative of HQ, was detected in 7% of the samples, while 4- phenoxy phenol appeared in 5% of the samples. Kojic Acid (KA) was found in 19% of the samples. Additionally, 4-butyl resorcinol, a potent tyrosinase inhibitor, was present in 59% of the samples, and 4-hydroxybenzoic acid, a common preservative, was detected in 43%. Heavy metals results provided a comprehensive insight into the presence of thirteen heavy metals (As, Cd, Cu, Cr, Co, Fe, Mn, Ni, Pb, V, Zn, Se, and Sb). Most samples contained multiple heavy metals, indicating a wide spread of occurrence across the tested products. Lead (Pb) was detected in forty-six samples (79.3%) at concentrations ranging from 0.001-15.41 ppm. Notably two samples, Chadni whitening cream and Rico had high concentrations at 3.71 ppm and 15.42 ppm respectively, above the threshold limit of 0.5 ppm. Mercury results were in the majority (85.75%) of samples exceeded the 1ppm safety threshold set by regulatory bodies. Products from Pakistan, China, India, and other countries were found to contain high mercury levels, with two samples, Omni Gold and Sandal, exhibiting exceptionally high concentrations. Notably, 55.4% of samples had mercury below 10 ppm, while 41.1% had levels in the tens to hundreds of ppm. Health risk assessment results indicated that hazard quotient levels were low, and the carcinogenic risk (CR) values for all detected metals were less than 1. Chromium (Cr) and Nickel (Ni) were the primary contributors to higher CR values. These findings highlight significant concerns regarding the presence of harmful substances in face creams, emphasizing the need for rigorous regulatory oversight. The health risk results for hydroquinone showed that seven samples, including Natural Glow Carotone (4.68 x 10⁻⁵), Coco Pulp (4.30 x 10⁻⁵), and Light & Bright (3.42 x 10⁻⁵), exceeded the CR threshold of 1 x 10⁻⁶ - that is, more than 1 in 1,000,000 persons was likely to develop cancer from continuous use of hydroquinone over the calculated period (70 years). These elevated CR values suggest a carcinogenic risk, with Natural Glow Carotone posing the highest risk among the tested samples. The hazard quotient values for creams containing mercury (ranging from 3.8 x 10−4 to 1.17 x 10−1 (0.117), were all below 1, indicating that the mercury content in these samples is unlikely to pose significant non-cancer health risks. Carcinogenic risk (CR) was below 1 x 10⁻⁶ for twenty -five samples (44%). However, thirty-three samples (56%) exceeded the cancer risk threshold of 1 x 10⁻⁶. Products with CR values above this threshold are considered unsafe for regular use, as they may expose consumers to potentially harmful levels of carcinogenic substances over time. Nine of the creams that contained hydroquinone also contained mercury. The calculated carcinogenic risk (CR) for hydroquinone was comparable higher than those of mercury, implying a greater contribution from hydroquinone to the overall risk from using the cream.
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    Open Access
    Folliculitis keloidallis nuchae severity score: development and reliability assessment
    (2020) Nyika, Dennias Toderai; Khumalo, Nonhlanhla; Isaacs, Thuraya
    Background: Folliculitis keloidalis nuchae (FKN) is a chronic inflammatory condition that targets the hair follicle, leading to keloidal scarring and alopecia. The absence of a severity scoring tool for FKN limits objective assessment of disease progression and response to treatment. Objectives: To develop and test the reliability of a severity scoring tool for FKN. Methods: The tool was developed based on lesion type, number, size and distribution on the scalp. An initial pilot period with 2 assessors was followed by the main study that used 78 anonymised and standardised clinical photographs of the back of the scalp. The participants were selected from an ongoing case control study of FKN. The assessors could allocate disease severity in one of 14 categories (with/without inflammation). However, inflammation (especially erythema) can be missed in photographs of pigmented skin. Thus, two groups of analysis were conducted first with all 14 and again with 8 categories (i.e. excluding inflammation). Assessors were 4 dermatology consultants and 7 registrars, who all independently scored the same anonymised and standardised photographs on two separate occasions, 2 weeks apart. Results: Inter-observer standard errors were higher with the 14-category compared to the 8- category analysis for both consultants and registrars. The intraclass correlation coefficient for registrars improved from poor [0.46 (0.36 -0.56)] to good [0.74 (0.68- 0.80)] with 14 compared to 8-categories, but stayed the same for consultants [0.82 (0.76 – 0.88) versus 0.81 (0.75 – 0.87)]. Limitations of the study were the use of clinical photographs instead of live participants and the problem that the signs of inflammation may be particularly difficult to judge in pigmented skin. Conclusion: We developed a severity scoring tool with poor to good reliability which also highlighted the difficulty of perceiving inflammation from clinical photographs. This improved with the seniority of the observer. The 8-category analysis has good reliability for clinical photographs for both junior and senior staff. For live patient care and clinical trials the 14-category version is likely to be more useful, but requires validation.
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    Open Access
    Human T cell lymphotropic virus type 1- associated infective dermatitis in KwaZulu Natal, South Africa
    (BioMed Central Ltd, 2013) Hlela, Carol; Graham, Natalie; Bhigjee, Ahmed; Taylor, Graham; Khumalo, Nonhlanhla; Mosam, Anisa
    BACKGROUND: The Human T cell lymphotropic virus type 1 (HTLV-1)-associated infective dermatitis (IDH), is a chronic relapsing dermatitis which usually presents in children older than 2years. A total of 300 cases have been reported worldwide (Latin America, the Caribbean and only 5 from Senegal). Neither IDH, nor its complications have been reported from the rest of Africa. We aimed to examine the clinical and aetiological characteristics of IDH in a cohort of South African children. METHODS: Attendees at the dermatology clinic at King Edward VIII Hospital, Durban underwent clinical examination. After obtaining consent those suspected of IDH had specimens taken for blood counts, immunoglobulins, serum protein electrophoresis, viral studies (including genotyping), skin swabs and stool examinations. RESULTS: Nineteen of 60 suspected cases recruited over 3years met the diagnostic criteria for IDH. The male-to-female ratio was 1:2; mean age 8years (range 0.7 to 15). Dermatitis mostly affected the scalp (78.9%) and axilla (73.7%); fewer children had nasal crusting (47.4%). Mean Ig A, IgG and IgM were raised, at 3.52g/l, 22.6g/l and 1.38g/l, respectively. The median CD4 cell count was 1958 cells/mm3. Viral genotyping of all tested samples were positive for the Cosmopolitan, Subtype A (HTLV-1a). CONCLUSIONS: IDH is a distinct entity which also affects South Africans. Our patients were older at presentation and the majority did not present with nasal crusting as has been described in other countries.
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    Interdependence between geometric, tensile and chemical bond behaviours of untreated hair fibres
    (2020) Cloete, Elsabe; Ngoepe, Malebogo; Khumalo, Nonhlanhla
    To date, an accurate understanding of the dynamics between the fibre's inherent geometric, mechanical and biological characteristics is deficient, affecting the reliability and robustness of hair data. There is also insufficient scientific clarity on the behaviour of curly hair, as most of the conclusions have been drawn from studies focusing on straight fibres. This research project aimed at gaining a more accurate understanding of the interrelationships between fibre curliness, strength and chemical bonding. In the current understanding of hair mechanics, curly fibres are considered to have a lower tensile strength than straight fibres. Furthermore, the current understanding of hair fibres does not associate hydrogen bonding with fibre shape. During experimentation, inadvertent observations suggested that current tensile methods ignore an important component of hair strength in curly fibres, and that hydrogen bonding supports fibre curliness. Intensive scrutiny of these observations led to fundamental contributions to the understanding of curly hair. Research tools included tensile, geometric, image, (FTIR) spectroscopic assessments, regression modelling and multivariate statistical analysis. Through this research, the role of hydrogen bonding in fibre curliness has been established. A theory is presented about extraordinary hydrogen bonds and the existence of hydrogen bond networks across the fibre matrix of curly hair. The theory has been substantiated experimentally via FTIR and weight measurements. The research also established the importance of the preelastic tensile region for curly fibres. It was clearly demonstrated that tensile strength of hair fibres is not only dependent on Young's modulus, but also on the fibre's inherent viscoelasticity, which appears to be important in curly fibres but becomes negligible with loss of curl. A model, developed from experimental observations and insights from similar biological fibres, is also presented. The model gives insights into ultrastructural changes at the early onset of fibre elongation. It also demonstrates the association between viscoelasticity and hydrogen bond networks. Taking this into consideration, a constitutive equation, developed to determine hair fibre strength accurately, is also presented in this work. This work does not replace current fibre curvature theories, but provides additional insights into hair shape, and therefore presents a fundamental contribution to curvature in human hair. It also highlights the shortcomings of current instrumentation methods that contribute to inaccurate conclusions regarding the strength of curly fibres.
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    Quantification of pyrethroid and organophosphate metabolites in hair and urine samples of rural children from the Western Cape, South Africa using the two-dimensional gas chromatography-mass spectrometry
    (2025) Mugari, Mufaro; Dalvie Aqiel, Mohamed; Khumalo, Nonhlanhla
    Extensive use of organophosphates (OPs) and pyrethroids (PYRs) in the Western Cape's farming regions raises significant health concerns, particularly for children, whose developing bodies are more susceptible to the toxic effects of these chemicals. Regular monitoring of these insecticide concentrations in human biological matrices is essential to identify and minimize exposure risks. However, existing chromatography monitoring systems encounter challenges such as sensitivity and matrix effects when quantifying these compounds in complex biological samples. This study evaluated a two-dimensional gas chromatography coupled with a time-of-flight mass spectrometry (2-D GCxGC TOF/MS) based method, specifically designed for analysis of compounds in complex matrices such as urine and hair matrices. Furthermore, the 2D GCxGC TOF/MS was available in-house and proved to be less expensive than outsourcing an analysis using a triple quadrupole mass spectrometer. The development and validation of the analysis method involved several key steps. First, the extraction techniques were optimized, utilizing QuEChERS for urine samples and solid-liquid extraction for hair samples. This was followed by the fine-tuning of metabolite derivatization using N-tert- butyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA) to ensure efficient analysis. The choice of extraction methods was tailored to each sample type to maximize recovery and data reliability. Additionally, MetaboAnalyst 5.0 was used on non-targeted analysis data, offering insights into exposure variations and the effects of OP and PYR exposure on metabolic pathways. Both targeted and non-targeted analysis was performed using 2-D GCxGC TOF/MS. The study focused on children from farming areas in Grabouw and the Hex River Valley, monitored over two cycles (2017 and 2018). The developed and validated method was applied to urine samples (n = 122) collected from 61 participants and hair samples (n = 176) collected from 176 children. Alongside these biological samples, detailed demographic, geographic, and agricultural practice data were collected to provide a comprehensive context for the analysis. The 2-D GCxGC TOF/MS technique demonstrated a limit of detection (LOD) ranging from 0.19 to 0.89 ng/mL and a limit of quantification (LOQ) ranging from 0.58 to 2.69 ng/mL for the targeted metabolites (dimethylphosphate-DMP, diethylphosphate-DEP, dimethylthiophosphate-DMTP, diethylthiophosphate-DETP, and 3-phenoxybenzoic acid-3PBA). Additionally, the method detection limit (MDL) for urine samples were between 0.16- 5.96 ng/mL and MDL for hair were between 2.27- 99.45 pg/mg. Although the LOD and LOQ in this study were higher than those typically reported using liquid chromatography-mass spectrometry (<0.1 ng/mL), the method effectively detected OP and PYR exposure levels in children from farming areas in the Western Cape, where agricultural activities lead to elevated exposure. Although extraction recoveries in this study were generally above 50%, the results remained below the typical range of 70-130 %. This could be due to complexity of polar metabolites within the urine matrix likely contributed to inefficient interactions with derivatization agents, additionally, the derivatization process also altered the properties of the analytes and matrix components, further reducing recovery rates. Ethyl acetate proved to be the most effective solvent for enhancing extraction and detection rates. The method was applied to the samples and the analysis showed OP and PYR metabolites in 50% of urine samples and 38% of hair samples. The method's robustness was demonstrated by its ability to detect and quantify metabolites at low levels. Universal exposure to OPs and PYRs among the children was indicated, likely driven by seasonal and agricultural practices. Notable findings included the detection of DMP, DEP, DETP, and 3-PBA at varying concentrations, suggesting episodic exposure events. The integration of MetaboAnalyst 5.0 with 2-D GCxGC TOF/MS provided a detailed examination of the metabolic pathways affected by insecticide exposure which revealed alterations in amino acid concentrations including glycine, serine, and threonine, suggesting potential disruptions in protein synthesis, energy metabolism and the nuanced biochemistry of human exposure to OPs and PYRs. Furthermore, non-targeted analysis revealed subtle shifts in the abundance of various metabolites involved in oxidative stress response and xenobiotic metabolism pathways, providing insights into the biochemical effects of OP and PYR exposure in children. The study demonstrated the efficiency of 2-D GCxGC TOF/MS in detecting and quantifying OP and PYR metabolites in biological samples, highlighting its potential for broader environmental and health research. The findings suggest potential geographical and agricultural practice-based variations in exposure levels, warranting further investigation into the nature of OP and PYR exposure in the region. Furthermore, the integration of analytical tools like MetaboAnalyst 5.0 enhanced the understanding of the metabolic impacts of OP and PYR exposure thus prepares for further research groundwork in this crucial field. This supports the need for customized biomonitoring strategies to manage the health risks of insecticide exposure in vulnerable groups, especially children. Continuous monitoring and advanced analytical methods are essential for safeguarding these at-risk populations.
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    The use of hair detect (and monitor) chronic hyperglycaemia
    (2018) da Silva, Sian-Ailin; Van Wyk, Jennifer; Khumalo, Nonhlanhla
    Background: Diabetes mellitus is a major public health problem resulting in about 5 million deaths per year. This metabolic disorder is characterized by hyperglycaemia, which results in debilitating and life-threatening complications. It is, therefore, vital for diabetics to monitor and control their blood glucose levels in order to keep them below 7mmol/L while fasting and below 9mmol/L after meals. Chronic estimates of glucose control of 8-12 weeks are obtained using glycated haemoglobin A1 (HbA1c). Non-invasive, less expensive methods of monitoring long term glycaemic control may be useful. Since scalp hair consists of about 80% protein, which is subject to non-enzymatic glycation, and growing hair has a rich blood supply exposing it to free glucose, it is likely that hair can be used as an alternative substrate for monitoring chronic hyperglycaemia. Subjects and Methods: Scalp hair and a blood samples (for HbA1c) were collected from 46 diabetic and 46 healthy control subjects. There were 26 diabetic adults (30-70 years), recruited from the outpatient clinic at Groote Schuur hospital and 20 children (7-18 years) recruited from the diabetic clinic at the Red Cross children's hospital. There were 29 healthy control adults (26-65 years) and 17 children (7- 17 years) recruited from the Groote Schuur and Red Cross hospitals respectively. History of chemical hair treatment was recorded for each participant. Hair samples were washed using 1% sodium dodecyl sulphate and analysed using Fourier transform infrared- attenuated total reflection (ATR-FTIR) spectroscopy. Spectra were analysed using statistical software (SIMCA, Umetrics) to determine whether the hair of diabetics was distinguishable from hair of healthy controls as well as whether spectra correlated with HbA1c levels of participants. Hair amino acid concentrations were also analysed as it is known that circulating amino acid concentrations are altered in people with diabetes. Results and discussion: The Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) models between spectra obtained from hair of diabetic participants and spectraobtained from control hair show good separation and predictive ability. When ATR-FTIR spectra were analysed in four groups: children with natural hair, adults with natural hair, adults with dyed hair and adults with relaxed hair, the models yielded predictive abilities (Q2(cum)) ranging from 0.753 to 0.85 and classification abilities ranging from 97.97 to 100%. This demonstrated excellent separation and predictive ability for controls vs. diabetics. Partial least squares (PLS) revealed a good correlation between hair FTIR spectra and participant HbA1c levels (R2 ranging between 0.8067 and 0.9296). These results demonstrated the possibility to use ATR-FTIR alongside multivariate data analysis to detect hyperglycaemia and monitor blood glucose levels via prediction of HbA1c levels from the hair spectra. Amino acid analysis supported the OPLS-DA classifications, as the largest differences were seen between age and chemically treated hair groups. Amino acid results reinforced the necessity to classify spectra into groups in order to distinguish between hair spectra from diabetics or controls, as well as to predict HbA1c. Twelve amino acids (Asp, Glu, Pro, Gly, Met, Ile, His, Lys, Arg, Amm, Cys, Leu) were significantly different between hair from adults and children, eight amino acids (Ala, Cys, Val, Met, Lys, Amm, Ser, Tyr) were statistically significantly different between natural and dyed hair and only up to four amino acids (Gly, Val, Met, Ile in children or Val, Phe, Amm, Arg in adults) were significantly different between diabetic and control groups. Conclusion: There is a need for non-invasive means of monitoring chronic hyperglycaemia. This study demonstrated the ability to distinguish between the hair of diabetics and controls as well as the ability to predict HbA1c levels from hair using ATR-FTIR. However, factors such as age and chemical treatment, which affect the chemical properties of hair, like amino acid levels, should be considered first. This would lead to promising prospects for long term blood glucose monitoring, due to the ability to estimate hair growth rate, and greater insights into the timing and development of diabetic complications. ATR-FTIR relatively simple to use, requires minimal sample preparation and does not require the use of expensive consumables. This technology could, potentially, be adapted into a primary health point of care or home screening or monitoring device for long-term hyperglycaemia, which would assist in early detection and preventing the progression of debilitating complications.