Diaryl pyrrolone fluorescent probing strategy for Mirabegron determination through condensation with ninhydrin and phenylacetaldehyde: Application to dosage forms, human urine and plasma.

Mirabegron (MRB) is a ß3-adrenoceptor agonist used for managing overactive bladder syndrome. A cost-effective, environmentally friendly, and highly sensitive spectrofluorimetric method was suggested to serve the purpose of quantifying MRB in its pure state, pharmaceutical tablets, spiked human plasma and urine, and testing content uniformity. In the present study, ninhydrin and phenylacetaldehyde react with the amino group moiety of MRB in Teorell-Stenhagen buffer (pH 7.5) to generate a strongly fluorescent diaryl pyrrolone compound that emits fluorescence at a wavelength of 477 nm upon excitation at 385 nm. The obtained calibration curve showed a linear relationship with a high correlation coefficient (r = 0.9997) in the concentration range of 0.25 to 5.0 µg mL-1. Limits of detection (LOD) and quantitation (LOQ) were 0.082 and 0.248 µg mL-1 respectively. The procedure was verified in accordance with the ICH guidelines. The suggested approach could be utilized for the selective analysis of MRB in its pharmaceuticals, either containing a single drug or co-formulated with solifenacin succinate. The greenness of the suggested method was confirmed using different green analytical metrics.

Further research into alternative carrier solvents for the detection of latent fingermarks.

A number of solvents, (Solstice PF, Opteon SF33 and Amolea AS-300), are compared to the recommended carrier solvent of HFE7100 for the ninhydrin and 1,2-indandione formulations. As the supply of HFE7100 will cease by the end of 2025, suitable alternatives are required in the short-term to ensure the detection of latent fingermarks on porous surfaces is still effective. Although these solvents, with the exception of Amolea AS-300, are classified as per- and polyfluoroalkyl substances (PFAS); they are not classed as hazardous. The alternatives in this study have a low global warming potential and atmospheric lifetime and are volatile, non-flammable and non-ozone depleting, in addition to other desirable properties such as a high wetting-index. During Phase 2 trials with deposited fingermarks, HFE7100 provided the best performing results followed by Opteon SF33, Solstice PF and Amolea AS-300. A significant difference with a negligible effect size was observed for ninhydrin formulations (p-value 0.00179; ε2 0.00418) while a significant difference with a weak effect size was observed for 1,2-indanedione formulations (p-value 2.095 ×10-10; ε2 0.0167). Furthermore, HFE7100 provided the least ink diffusion and the brightest 1,2-indanedione luminosity (significant difference with a moderate effect size p-value 1.772 ×10-13; ε2 0.0434) but the HFE formulation turned cloudy more quickly and needed regular replacements. Phase 3 pseudo-operational trials of 100 porous items followed a similar trend whereby HFE7100 formulations detected the highest number of marks followed by Opteon SF33 and Solstice PF. Although HFE7100 is still the best performing carrier solvent, this study demonstrates that, in the short-term, Opteon SF33 and Solstice PF may have potential as non-flammable replacement carrier solvents while developing the long-term goal of solvent-less methods.

Development of sensitive spectrofluorometric approach based on formation of pyrrolidone derivative for determination of linagliptin through condensation reaction with ninhydrin and phenylacetaldehyde: Application to content uniformity and real plasma.

The new drug linagliptin belongs to the class of dipeptidyl peptidase-4 enzyme inhibitors. Linagliptin is used to treat type 2 diabetes and is taken orally either alone or in combination with other drugs. In this instance, a new, simple, and economical technique for analyzing linagliptin was developed by the effective use of a pyrrolidone derivative. The primary amine group of linagliptin permits its condensation with ninhydrin (0.14% w/v) to produce a fluorescent product in the presence of phenylacetaldehyde (0.02% v/v). All experimental parameters were carefully examined and adjusted in order to monitor the generation of the pyrrolidone derivative at excitation and emission wavelengths of 385 and 475 nm, respectively. The calibration graph was made by plotting the intensity of the fluorescence in relation to linagliptin concentration. A significant linearity was found for values ranging from 20 to 460 ng/mL. The process's validity has been verified by a thorough assessment of the instructions provided by the International Conference on Harmonization (ICH). The results indicate excellent uniformity with a reference method, showing that there is no substantial difference in precision and accuracy. The proposed approach was utilized for determining linagliptin in real rat plasma successfully owing to its high sensitivity. Additionally, the proposed approach was evaluated using the Eco-Scale evaluation tool and showed a high degree of eco-friendliness (86/100).

The laboratory perspective: Confirming the integrity of fingermark enhancement reagents.

Validation is particularly important in forensic science. Each process utilised by a forensic laboratory will undergo validation to ensure it is functioning as expected. Some pieces of equipment (hardware) will be simple to validate, and this will be conducted regularly. However, many commonly employed fingermark enhancement reagents, such as ninhydrin and 1,2-Indandione, do not have clearly defined and robust validation parameters. To that end, artificial fingermark solutions, particularly amino acid solutions, have previously been investigated as a controlled validation method, via deposition onto a substrate. Whilst amino acid reagents are important, there is also a wider group of latent fingermark constituents (targets) that require the use of other enhancement reagents within the forensic laboratory. The work presented herein seeks to expand on the concept of amino acid printing, by exploring the possibilities of depositing a sebaceous solution matrix, thereby facilitating the testing of reagents such as Oil-Red-O and Physical Developer. In addition, we present a method that extends capabilities beyond fingermark enhancement reagents for porous substrates, by developing a methodology that enables process validation of the cyanoacrylate fuming technique to be easily facilitated. These simple and effective solutions have the capacity to serve as a crucial process validation check within the laboratory validation workflow.

Designing stimuli-responsive upconversion nanoparticles based on a mimetic immunoassay for potential accurate diabetic nephropathy diagnosis.

Diabetic nephropathy (DN) is the most common microvascular complication associated with incurable diabetes. The gold standard diagnostic method for DN is based on the detection of proteinuria but it overlooks cases of non-proteinuria (NP-DN). To address this limitation, urinary sialic acid (SA) has been confirmed as an effective biomarker for various DNs. Herein, we constructed an ultrasensitive non-proteinuria assay platform to accurately diagnose DN within 20 min. This platform utilized the ninhydrin reaction between acidic ninhydrin and urinary sialic acid (SA) as an effective biomarker for various DNs. A compound with a maximum absorption peak at 470 nm was produced in this reaction and contributed to the fluorescence decrease of the blue-emission core-shell upconverting nanoparticles through the inner filter effect (IFE). By integrating the inner filter effect (IFE) with a mimetic immunoassay, the imperceptible color was converted into highly sensitive fluorescence signals. This protocol shows a stable and high sensitivity with a detection limit of 20 nmol L-1 and provides 100% positive prediction for urine samples, demonstrating its potential for clinical diagnosis and long-term monitoring of DN.

Development of a colorimetric sensor based on nanofiber cellulose film modified with ninhydrin to measure the formalin index of fruit juice.

In this research, a color sensor based on nanofiber cellulose film modified with ninhydrin was designed to measure amino acids and formalin index in fruit juice. For this purpose, three types of cellulose films with porosity of 5, 30 and 125 µm were used. These films were treated with standard solution of ninhydrin. The characteristics of modified films were investigated using Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) and X-ray Diffraction (XRD) tests. The color factors of the sensors (a and b) changed in the presence of amino acids and juice with different levels of formalin index. Therefore, the modified films with ninhydrin as a colorimetric sensor were calibrated using 7 types of amino acids and based on the formalin index of 4 types of juice. Then the sensors were used to measure the formalin index in 4 types of juice. The results showed that the sensors have relative selectivity towards methionine amino acid. The formalin index values calculated in the juices by the sensor were compared with the titration method as a reference method. All three types of sensors were able to detect formalin index. The results of the sensor performance verification showed that the sensors can measure formalin index in different juices with 95-98 % accuracy. These sensors showed fast sensitivity and selectivity to the amino acids in juice, also these sensors are safe and the measurement method is fast and simple.

A paper-based chemical tongue based on the charge transfer complex of ninhydrin with an array of metal-doped carbon dots discriminates natural amino acids and several of their enantiomers.

Simultaneous detection of multiple amino acids (AAs) instead of individual AAs is inherently worthwhile for improving diagnostic accuracy in clinical applications. Here, a facile and reliable colorimetric microfluidic paper-based analytical device (µPAD) using carbon dots doped with transition metals (Cr3+, Mn2+, Fe3+, Co2+, Ni2+, Cu2+, and Zn2+) has been provided to detect and discriminate 20 natural amino acids. To make the colourless metal-doped carbon dots suitable for colorimetric assays, they were mixed with ninhydrin to form a charge transfer complex. This optical tongue system, which was constructed by dropping mixtures of ninhydrin with a series of metal-doped carbon dots on a paper substrate in an array format, represented obvious but different colorimetric signatures for every examined amino acid. Since bovine serum albumin was used as a chiral selector reagent for synthesizing the CDs, the sensor device represented excellent selectivity to identify enantiomeric species of AAs. This is the first optical array device that can simultaneously discriminate AAs and several of their enantiomers. We employed various statistical and chemometric methods to analyze the digital data library collected by Image J software, including principal component analysis (PCA), linear discriminant analysis (LDA), and hierarchical cluster analysis (HCA). Twenty AAs could be well distinguished at various concentrations (10.00, 5.00, 2.50, and 1.25 mM). The colorimetric patterns were highly repeatable and were characteristic of individual AAs. Besides qualitative analysis, the designed µPAD-based optical tongue represented quantitative analysis ability, e.g., for lysine in the concentration ranges of 0.005-20.0 mM with a detection limit of 1.0 × 10-6 M and for arginine in the concentration range of 0.12-20.00 mM with a detection limit of 80.0 × 10-6 M. In addition, the binary, ternary, and quaternary mixtures of AAs could also be well recognized with this sensor.

The influence of storage conditions on fingermarks developed with 1,2-indanedione-ZnCl.

The most important class of reagents for the enhancement of fingermarks on porous surfaces are those that react with the amino acid constituents of fingermarks. Ninhydrin, DFO (1,8-diazafluoren-9-one), and 1,2-indanedione are the three most common techniques widely known in forensic laboratories for the visualization of latent fingermarks on porous surfaces. In 2012 the Netherlands Forensic Institute replaced DFO by 1,2-indanedione-ZnCl after an internal validation, just like an increasing numbers of laboratories. In 2003, Gardner et al. published an article showing that fingermarks treated with 1,2-indanedione (without ZnCl), that were stored in daylight only decreased by 20% in fluorescence in 28 days. However, during casework we observed that the fluorescence of fingermarks treated with 1,2-indanedione with ZnCl decreased more rapidly. In this study, the effect of various storage conditions and aging times on the fluorescence of marks after treatment with 1,2-indanedione-ZnCl were assessed. Latent printed fingermarks from the digital matrix printer (DMP) and natural fingermarks from a known donor were used. The results showed that storing fingermarks in daylight (wrapped and un-wrapped) drastically decrease (over 60% loss) in fluorescence in approximately three weeks. Storage of the marks in a dark environment (at room temperature, in the refrigerator or even in the freezer) resulted in a decrease in fluorescence of less than 40%. Our recommendation is to always store treated fingermarks with 1,2-indanedione-ZnCl in a dark environment and, if possible, photographing them directly (within 1-2 days after treatment) to minimize the decrease of fluorescence.

Development of Sٍٍensitive Spectrofluorimetric Methods for Determining Netilmicin Based on Selective Condensation Reactions of its Amine Moiety with each Acetylacetone/Formaldehyde and Ninhydrin/Phenylacetaldehyde Reagents.

Netilmicin is an aminoglycoside antibiotic used to treat infections caused by a broad spectrum of Gram-negative and Gram-positive bacteria and is pharmaceutically formulated in ophthalmic dosage forms. In this study, two spectrofluorimetric approaches were designed and developed to switch-on the fluorescence activity of NTC. The first method, or Hantzsch (HNZ) method, was relied on measuring the generated fluorescence intensity upon the condensation of NTC with acetylacetone and formaldehyde (Hantzsch reaction) at λemis=483 nm/λexcit=425.5 nm. While the second fluorometric method (NHD method) was relied on measuring the generated fluorescence intensity upon the condensation of NTC with ninhydrin/phenylacetaldehyde at λemis=482.2 nm/λexcit=385.8 nm. The reaction conditions for the two approaches were well investigated and optimized. The selectivity study for the methods was investigated by determining NTC in the presence of the co-formulated drug (dexamethasone) and pharmaceutical excipients. The validation for two approaches was performed based on ICH guidelines, and ranges of linearity were 0.1-1.2 and 1.5-6.0 µg/mL, while LOD values were 0.039 and 0.207 µg/mL for the HNZ method and the NHD method, respectively. Finally, NTC has been determined in different ophthalmic preparations by the proposed approaches with adequate recovery values.

Quantifying DNA loss in laboratory-created latent prints due to fingerprint processing.

Fingerprints, which are associated with touch samples, typically contain a limited amount of DNA. The amount of available DNA can be further reduced when the same touch samples undergo fingerprint processing [1]. The fingerprint development process consists of high-powered lighting (inherent luminescence and UV light) and chemical compounds (ninhydrin, black powder, cyanoacrylate, and rhodamine 6 G) which could reduce DNA quality and quantity. Therefore, forensic scientists often must select one type of analysis over the other due to the destructive nature of processing. DNA and latent fingerprinting are both useful sources for identification, although both can produce partial results. A partial DNA profile may only contain a few alleles, limiting the ability to identify a potential suspect to perform comparisons. A partial fingerprint generally means that only a very small part of the fingerprint is present, which makes comparisons difficult. Because partial results are common, combining data from both fingerprinting and DNA analysis would increase the confidence of an identification of a person. Significant research has been performed to determine if a DNA profile can be obtained from latent processed fingerprints; however, there has yet to be research done in a standardized manner. In this study, we used standardized mock "fingerprints" in order to reduce fingerprint DNA variability and specifically focused on DNA quantitation after each step in the fingerprinting process. Results suggest that latent print processing techniques used on non-porous surfaces (plastic, duct-tape, metal, and rubber) do not affect DNA quantity or quality. In contrast, ninhydrin, a chemical used for processing fingerprints present on porous surfaces (wood and paper), significantly reduced DNA recovery. Together these results suggest that DNA can still be performed on latent print processed items, unless ninhydrin has been used.

Survey reaction of 2-amino-N-(aryl) benzimidamides with ninhydrin in different conditions and investigation of reaction mechanism using density functional theory (DFT).

In this investigation, firstly, 1-(2-amino-phenyl)-N-(aryl) methane diamine derivatives were synthesized by reaction of 2-aminobenzo nitrile with aromatic amines in the presence of aluminum chloride as the catalyst. Then, the reaction of these intermediates with ninhydrin in different conditions was investigated. The reaction between ninhydrin and 2-amino-N'-(aryl) benzimidamide derivatives in water as solvent under reflux conditions resulted in the synthesis of diazepine derivatives. The same results were obtained when the reaction was done in EtOH and in the presence of a few drops of sulfuric acid at room temperature. Also, this reaction was carried out in ethanol as solvent without the presence of sulfuric acid at room temperature which resulted in the synthesis of spiro [indene-2,2'-quinazoline] derivatives. And finally, the reaction was carried out in ethanol as solvent without the presence of sulfuric acid at the reflux conditions which resulted in the synthesis of isoquinolino-quinazoline derivatives. These N-heterocycles compounds are important biologically. Mild reaction conditions, simple procedure and purification and also product diversity with changing conditions are important advantages of this method. Also, to better understanding reaction mechanism on the condensation reactions of 2-amino-N-(aryl) benzimidamides with ninhydrin in different conditions, density functional theory (DFT)-based quantum chemical methods have been applied. Calculated atomic charges suggest that the C-1 (+ 0.54 a.u.) center of ninhydrin is a better electrophile than C-2 (+ 0.42 a.u.) center.

Enzyme-free colorimetric sensor based on molecularly imprinted polymer and ninhydrin for methamphetamine detection.

Digital image colorimetry was applied to introduce a rapid, portable, and non-enzymatic test for methamphetamine measurements in urine. Imprinted polymer was synthesized in a simple, low-cost process and utilized for selective extraction of analyte from the sample in combination with the well-known ninhydrin color test. Applying the digital camera on a mobile phone, RGB basic color data were obtained, and calibration curves were developed for different concentrations of methamphetamine. Optimization of the test condition was carried out by changing some effective parameters such as extraction time and pH. The results were compared with some similar structural compounds indicating great potential for use as a selective and semi-quantitative field test for this drug. An acceptable linear range (5-100 µM) and detection limit (1.44 µM) as well as good agreement with the reference method, makes this fast portable method, an easy and reliable test for the analysis of methamphetamine in biological samples.

Diastereo- and regioselective petasis aryl and allyl boration of ninhydrins towards synthesis of functionalized indene-diones and dihydrobenzoindeno-oxazin-ones.

Petasis aryl and allyl borations were accomplished using substituted ninhydrins, boronic acids or 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane and 1,2-aminophenols in Hexafluoroisopropanol (HFIP) without any catalysts to synthesize different aryl and allyl derivatives of ninhydrins. The nature of substitution in the boronic acids and 1,2-amino phenols was the key factor in determining the diastereo-regioselectivity and the type of product distributions. The products were isolated and characterized by HMBC, HSQC, 1H, 13C NMR experiments and X-ray single crystallographic analysis. A probable reaction pathway involves in situ formation of acyclic and cyclic ninhydrin-amino alcohol adducts, with the positioned hydroxyl group determining the stereo-regioselective outcome via tetracoordinated boron intermediates. A metal free diastereo- and regioselective Petasis aryl and allyl boration of ninhydrins.

Preliminary evaluation of Solstice® PF as a replacement carrier solvent for Australian fingermark detection.

HFE-7100 is a routine carrier solvent in amino acid-sensitive fingermark detection reagents such as ninhydrin and 1,2-indanedione/zinc chloride (IND/Zn). However, a potential EU ban on hydrofluoroethers may require reformulation of these treatments worldwide. Solstice® PF has shown promise as a replacement for HFE-7100 in the United Kingdom. However, the performance (and hence optimal formulation) of IND/Zn is impacted by differences in climate and substrate composition, necessitating assessments under local conditions for different regions. We present a series of preliminary investigations in an Australian context, using the IND/Zn formulation used by Australian forensic service providers. The general performance of Solstice® PF-based IND/Zn was comparable to that using HFE-7100 on three substrate types, three ageing periods (1, 7 and 30 days) and 5 donors. However, slight differences in colour and luminescence intensity, as well as increased ink diffusion, suggest chemical interactions with other reagent components that may affect stability. Specifically, Solstice® PF-based reagent formed a precipitate within a month of storage, though this did not affect performance over a 4 month period. HFE-7100-based IND/Zn was found to be marginally more effective than Solstice® PF when applied to incidental fingermarks. These results indicate that Solstice® PF is a satisfactory alternative carrier solvent to HFE-7100 in an Australian context, though users should be aware of possible limitations regarding compatibility with other evidence components (particularly inks) and shelf-life.

Synthesis of 1,3,5-trisubstituted pyrazoles via 1,3-dipolar cycloaddition of nitrile imines with ninhydrin-derived Morita-Baylis-Hillman carbonates.

An effective synthetic method for 1,3,5-trisubstituted pyrazoles via 1,3-dipolar cycloaddition reaction has been developed. This reaction could smoothly proceed between ninhydrin-derived Morita-Baylis-Hillman carbonates and nitrilimines to provide a wide scope of differently substituted pyrazoles in high yields (up to 95%). In addition, the reaction mechanism was also proposed to explain its regioselectivity.

A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent.

An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.

Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea.

Thermal stress is considered one of the main causes of mass scleractinian coral degradation; however, it is still unknown how corals can adapt to future global warming. In this study, 11 strains of coral-associated Flavobacteria were shown to produce zeaxanthin, a carotenoid antioxidant, which may help coral holobionts to alleviate thermal stress. In addition, a novel zeaxanthin-producing Flavobacterium, designated R38T, was identified using polyphasic taxonomy. Although strain R38T shared a maximum 16S rRNA gene sequence similarity of 93% with Mesoflavibacter aestuarii KYW614T, phylogenetic analyses based on whole genome and 16S rRNA gene sequences revealed that strain R38T forms a distinct branch in a robust cluster composed of strain R38T and Leptobacterium flavescens KCTC 22160T under the family Flavobacteriaceae. Strain R38T exhibited average nucleotide identities of 70.2% and 72.5% for M. aestuarii KYW614T and L. flavescens KCTC 22160T, respectively. The only detected respiratory quinone was menaquinone 6 (MK-6). The genomic DNA G + C content was 33.2 mol%. The major polar lipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The major cellular fatty acids were iso - C15: 0, iso - C15: 0 ω6c, C16:2 DMA, and C13:1 ω3c. The distinct biochemical, chemotaxonomic, phylogenetic, and phylogenomic differences from validly published taxa suggest that strain R38T represents a new species of a new genus, for which Prasinibacter corallicola gen. nov., sp. nov. is proposed. The type strain R38T (= MCCC 1K03889T = KCTC 72444T).

Organocatalytic Regio- and Enantioselective [3 + 2]-Annulations of Ninhydrin-Derived Morita-Baylis-Hillman Carbonates with 3-Methyleneoxindoles.

A type of Morita-Baylis-Hillman (MBH) carbonates has been developed from ninhydrin. These MBH carbonates have been successfully employed as 3C-synthons in the organocatalytic asymmetric [3 + 2]-annulations of the isatin-derived electron-deficient olefins, affording structurally diverse spirooxindoles in high yield with excellent stereoselectivity. In particular, the regioselectivity of MBH carbonates was controlled by the reaction partner, 3-methyleneoxindoles with carbonyl groups (R = ArCO), affording ß-selective products and 3-methyleneoxindoles with ester groups (R = CO2Me) furnishing γ-selective products. The representative scale-up reactions and transformation of product were examined. The reaction mechanism was expounded by control experiments.

Production of artificial fingermarks. Part I - Synthetic secretions formulation.

Fingermark variability is a critical parameter. To mitigate the effects of this variability, synthetic secretions in the form of simple mixtures of target compounds found in eccrine sweat have been described in the literature, but they are usually reactive towards only a minimal range of detection techniques. If this approach is acceptable for the production of single-technique test strips, such artificial secretions cannot be considered as reliable fingermark simulants because they do not reproduce the complex matrix that makes up real secretions. Research has shown that sebaceous and eccrine compounds are probably present simultaneously in fingermark residue in the form of an emulsion. This paper is the first part of a research project that aims at producing realistic artificial fingermarks containing an extensive range of eccrine and sebaceous compounds. This first study aimed to reproduce and compare two synthetic fingermark residues formulations and assess their potential to be used as fingermark simulants. Spot tests of the artificial secretions were deposited on paper substrates, and their reactivity with four common detection techniques was tested: 1,2-indanedione-zinc, ninhydrin, oil red O, and physical developer. Both formulations showed very good results when processed with the two amino acid reagents, as well as oil red O, and no obvious differences were observed between the two versions. The results obtained with the physical developer were inconsistent and demonstrated that the fundamental working principle of physical developer needs to be further understood. The results were extremely promising as they showed the potential of such reproducible artificial secretions to be used to assess an extensive range of detection techniques, which would be highly beneficial to guarantee better research and quality control in fingermark detection. The use of spot tests to deposit the simulant was shown to be unreliable and a more controllable and reproducible deposition method using an inkjet printer will be presented in the second part of this research.

DNA recovery after sequential processing of latent fingerprints on copy paper.

Forensic examiners must determine whether both latent fingerprint development and DNA profiling can be performed on the same area of an evidence item and, if only one is possible, which examination offers the best chance for identification. Latent fingerprints can be enhanced by targeting different components of fingerprint residues with sequential chemical treatments. This study investigated the effects of single-reagent and sequential latent fingerprint development processes on downstream DNA analysis to determine the point at which latent fingerprint development should be stopped to allow for DNA recovery. Latent fingerprints deposited on copy paper by one donor were developed using three sequential processes: 1,8-diazafluoren-9-one (DFO) â†’ ninhydrin â†’ physical developer (PD); 1,2-indanedione-zinc (IND-Zn) â†’ ninhydrin â†’ PD; and IND-Zn â†’ ninhydrin â†’ Oil Red O (ORO) â†’ PD. Samples were examined after the addition of each chemical treatment. DNA was collected with cotton swabs, extracted, quantified, and amplified. DNA yields, peak heights, number of alleles obtained, and percentage of DNA profiles eligible for CODIS upload were examined. DNA profiles were obtained with varying degrees of success, depending on the number and type of treatments used for latent fingerprint development. The treatments that were found to be the least harmful to downstream DNA analysis were IND-Zn and IND-Zn/laser, and the most detrimental treatments were DFO, DFO/laser, and PD. In general, as the number of treatments increase, the opportunities for DNA loss or damage also increase, and it is preferable to use fewer treatments when developing latent fingerprints prior to downstream DNA processing.