Heteronuclear Correlation Research Articles

Development of 19 F-detected 1,1-ADEQUATE for the characterization of polyfluorinated and perfluorinated compounds.

Polyfluorinated and perfluorinated compounds in the environment are a growing health concern. 19 F-detected variants of commonly employed heteronuclear shift correlation experiments such as heteronuclear single quantum correlation (HSQC) and heteronuclear multiple bond correlation (HMBC) are available; 19 F-detected experiments that employ carbon-carbon homonuclear coupling, in contrast, have never been reported. Herein, we report the measurement of the 1 JCC and n JCC coupling constants of a simple perfluorinated phthalonitrile and the first demonstration of a 19 F-detected 1,1-ADEQUATE experiment.

Expedited Nuclear Magnetic Resonance Assignment of Small- to Medium-Sized Molecules with Improved HSQC-CLIP-COSY Experiments.

Resonance assignment is a pivotal step for any nuclear magnetic resonance (NMR) analysis, such as structure elucidation or the investigation of protein-ligand interactions. Both 1H-13C heteronuclear single quantum correlation (HSQC) and 1H-1H correlation spectroscopy (COSY) two-dimensional (2D) experiments are invaluable for 1H NMR assignment, by extending the high signal dispersion of 13C chemical shifts onto 1H resonances and by providing a high amount of through-bond 1H-1H connectivity information, respectively. The recently introduced HSQC-CLIP(Clean In-Phase)-COSY method combines these two experiments, providing COSY correlations along the high-resolution 13C dimension with clean in-phase multiplets. However, two experiments need to be recorded to unambiguously identify COSY cross-peaks. Here, we propose novel variants of the HSQC-CLIP-COSY pulse sequence that edit cross-peak signs so that direct HSQC responses can be distinguished from COSY relay peaks, and/or the multiplicities of the 13C nuclei are reflected, allowing the assignment of all the peaks in a single experiment. The advanced HSQC-CLIP-COSY variants have the potential to accelerate and simplify the NMR structure-elucidation process of both synthetic and natural products and to become valuable tools for high-throughput computer-assisted structure determination.

Incorporation of 5-Nitroisatin for Tailored Hydroxyapatite Nanorods and its Effect on Cervical Cancer Cells: A Nanoarchitectonics Approach

Hydroxyapatite (HAP) have been widely used as bone implants for its biocompatibility, and bioactivity. Other approaches like scaffold development, drug delivery and nanomaterials preparation are explored recently. Herein, hydroxyapatite incorporated with 5-nitroisatin (HAP-5 N) were prepared and tested for its anticancer potential. The prepared HAP-5 N were found to have rod like structures as seen using scanning electron microscopic analysis. The structural elucidation of 5-nitroisatin were confirmed by performing 2D heteronuclear single quantum correlation experiment. The synthesized HAP-5 N were assessed for its possible role as an anticancer agent against cervical cancer (SiHa) cells. From the study, it was found that HAP-5 N is efficient in inhibiting the growth of cervical cancer cell lines.

Consistency of a dialyzable leucocyte extract manufactured at GMP facilities by nuclear magnetic resonance spectroscopy

The present work describes the development and validation of a first report including several non-invasive NMR schemes to identify parameters as local chemical environments, homo- and heteronuclear site-specific spin correlations, diffusion coefficient-dependent polydispersity indexes and quantification of identified peptide entities that composes a commercial human Dialyzable Leucocyte Extract (DLE), Transferon, an oral liquid formulation of low-molecular-weight peptides. The above parameters were useful indicators to verify reproducibility, consistency and homogeneity among the DLE batches manufactured at Good Manufacturing Practice (GMP) facilities and for batch-releasing purposes in a quality control laboratory. The results showed that peptide identity of the DLE is represented with both high reproducible one-dimensional proton spectra and diffusion coefficient distributions that predicts in turn a weight-average molecular weight of around 6.7-7.4 kDa and a mean polydispersity index of 1.13. The obtained NMR peptide fingerprint of the analyzed DLE allowed to i) confirm its structural homogeneity by line-shape analysis, ii) identify and quantify its peptide content within the total solution with qNMR methods iii) to confirm the robustness of the technique as a feasible alternative for routine analysis of Natural or non-Natural Complex Drugs, such as DLEs.

Elucidating the Location of Cd2+ in Post-synthetically Treated InP Quantum Dots Using Dynamic Nuclear Polarization 31P and 113Cd Solid-State NMR Spectroscopy

Indium phosphide quantum dots (InP QD) are a promising alternative to traditional QD materials that contain toxic heavy elements such as lead and cadmium. However, InP QD obtained from colloidal synthesis are often plagued by poor photoluminescence quantum yields (PL-QYs). In order to improve the PL-QY of InP QD, a number of post-synthetic treatments have been devised. Recently, it has been shown that InP post-synthetically treated with Lewis acid metal divalent cations (M-InP) exhibit enhanced PL-QY; however, the molecular structure and mechanism behind the improved PL-QY are not fully understood. To determine the surface structure of M-InP QD, dynamic nuclear polarization surface-enhanced nuclear magnetic resonance spectroscopy (DNP SENS) experiments were employed on a series of InP magic size clusters treated with Cd ions, InP QD, cadmium phosphide (Cd₃P₂) QD, and Cd-treated InP QD (Cd–InP QD). With the use of DNP SENS, we were able to obtain the 1D ³¹P and ¹¹³Cd NMR spectra, ¹¹³Cd{³¹P} rotational-echo double-resonance (REDOR) NMR spectra, and ³¹P{¹¹³Cd} dipolar heteronuclear multiple quantum correlation (D-HMQC) sequence. Changes in the phosphide ³¹P chemical shifts after Cd treatment provide indirect evidence that some Cd alloys into the sub-surface regions of the particle. DNP-enhanced ¹¹³Cd solid-state NMR spectra suggest that most Cd ions are coordinated by oxygen atoms from either carboxylate ligands or surface phosphate groups. ¹¹³Cd{³¹P} REDOR and ³¹P{¹¹³Cd} D-HMQC experiments confirm that a subset of Cd ions are located on the surface of Cd–InP QD and coordinated with phosphate groups.

Multinuclear Nuclear Magnetic Resonance Spectroscopy Is Used to Determine Rapidly and Accurately the Individual pK a Values of 2-Deoxystreptamine, Neamine, Neomycin, Paromomycin, and Streptomycin.

Unambiguous assignments have been made for each individual pKa value of the amino group and guanidine substituents on 2-deoxystreptamine, neamine, neomycin, paromomycin, and streptomycin by pH-titration evaluation of their 1H, 13C, and 15N (by 1H-15N heteronuclear multiple-bond correlation (HMBC) spectra) NMR chemical shifts (δXs) as the reporter nuclei. These data require minor revisions of the literature data in terms of the assignment order for neomycin and paromomycin. In situ titrations and NMR spectroscopy are shown to be a powerful combination for rapidly (minutes) obtaining each distinct pKa value of the similar amine and guanidine functional groups, which decorate aminoglycoside antibiotics.

Polymerized Luteolin Nanoparticles: Synthesis, Structure Elucidation, and Anti-Inflammatory Activity.

Luteolin is an anti-inflammatory flavonoid commonly found in many edible plants. The compound is popularly consumed as a supplement regardless of its poor water solubility (27.8 μg/mL at 25 °C) and low bioavailability. Here, mild one-pot polymerization of luteolin into water-dispersible nanospheres, with an average dry size of 234.8 ± 101.6 nm, an aqueous size distribution of 379.1 ± 220.5 nm (PDI = 0.338), an average ζ-potential of −36.2 ± 0.2 mV, and an 89.3 ± 4.8% yield, is described. The nanospheres consist of polymerized luteolin (polyluteolin) with a weight-average molecular mass of around 410000 Da. The chemical structure of polyluteolin is identified through 1H–1H correlated spectroscopy (COSY), 1H–13C heteronuclear single-quantum coherence (HSQC), and 1H–13C heteronuclear multiple-bond correlation (HMBC) NMR spectroscopic analyses of the oligomers, and a polymerization mechanism is proposed. Unlike luteolin that showed both dose-dependent anti-inflammatory activity and cytotoxicity when tested in lipopolysaccharide-stimulated macrophages, the polyluteolin nanoparticles possess dose-dependent anti-inflammatory activity without causing cell death even at high concentrations.

Determination of tautomeric preference of fenobam in solution by high-resolution NMR spectroscopy.

In this work, tautomeric preference of fenobam in solution was investigated by homonuclear and heteronuclear solution nuclear magnetic resonance (NMR) spectroscopy. 1 H-1 H nuclear Overhauser effect spectroscopy (NOESY) spectrum revealed that fenobam in liquid state exists exclusively in one of the two possible tautomeric structures, which was confirmed by 1 H-13 C HSQC and heteronuclear multiple bond correlation (HMBC) spectra. Moreover, difference between the two tautomeric structures was studied by theoretical calculations, which further proved the result obtained by the NMR experiments.

Light-Switchable and Self-Healable Polymer Electrolytes Based on Dynamic Diarylethene and Metal-Ion Coordination.

Self-healing polymer electrolytes are reported with light-switchable conductivity based on dynamic N-donor ligand-containing diarylethene (DAE) and multivalent Ni2+ metal-ion coordination. Specifically, a polystyrene polymer grafted with poly(ethylene glycol-r-DAE)acrylate copolymer side chains was effectively cross-linked with nickel(II) bis(trifluoromethanesulfonimide) (Ni(TFSI)2) salts to form a dynamic network capable of self-healing with fast exchange kinetics under mild conditions. Furthermore, as a photoswitching compound, the DAE undergoes a reversible structural and electronic rearrangement that changes the binding strength of the DAE-Ni2+ complex under irradiation. This can be observed in the DAE-containing polymer electrolyte where irradiation with UV light triggers an increase in the resistance of solid films, which can be recovered with subsequent visible light irradiation. The increase in resistance under UV light irradiation indicates a decrease in ion mobility after photoswitching, which is consistent with the stronger binding strength of ring-closed DAE isomers with Ni2+. 1H-15N heteronuclear multiple-bond correlation nuclear magnetic resonance (HMBC NMR) spectroscopy, continuous wave electron paramagnetic resonance (cw EPR) spectroscopy, and density functional theory (DFT) calculations confirm the increase in binding strength between ring-closed DAE with metals. Rheological and in situ ion conductivity measurements show that these polymer electrolytes efficiently heal to recover their mechanical properties and ion conductivity after damage, illustrating potential applications in smart electronics.

Synthesis and properties of fluorinated benzotriazole‐based donor‐acceptor‐type conjugated polymers via Pd‐catalyzed direct CH/CH coupling polymerization

AbstractFluorine substituted benzotriazole (BTz) units have shown great potential in improving various conjugated polymers (CPs)‐based optoelectronic devices' performance. Thus, it is highly expected for the establishment of simple, efficient, and inexpensive accesses to such polymers. In this paper, Pd‐catalyzed direct CH/CH coupling polymerization is first employed for the synthesis of 5,6‐difluorobenzotriazole containing π‐CPs, which fully avoids prefunctionalization of monomers. Under the optimized conditions, a series of donor‐acceptor‐type π‐CPs with excellent regio‐regularity are facilely obtained via the direct CH/CH coupling reaction of 5,6‐difluoro‐2‐(2‐hexyldecyl)‐2H‐BTz with different thiophene analogs. The chemical structure of the as‐synthesized polymers are confirmed by NMR technique including 1H NMR, 19F NMR, and the edited heteronuclear singular quantum correlation and heteronuclear multiple bond correlation spectra as well as comparative study on the same polymers obtained via Stille coupling and direct (hetero)arylation polymerization. Further, their optical properties, electrochemical properties, and thermal stabilities are also carefully explored by UV–Vis absorption spectra, cyclic voltammograms as well as thermogravimetric analysis, respectively. On the basis of density functional theory (DFT) simulation, the effect of alkyl substituents attached to thiophene units on the reactivity and optical performance of the resultant polymers is explained. Our protocol exhibits remarkable advantages in synthetic simplicity, atom‐economy, high efficiency, and excellent regioselectivity of cross‐coupling, providing an alternative synthetic strategy for high‐performance optoelectronic materials.

Uncovering selective and active Ga surface sites in gallia-alumina mixed-oxide propane dehydrogenation catalysts by dynamic nuclear polarization surface enhanced NMR spectroscopy.

Gallia–alumina (Ga,Al)2O3(x : y) spinel-type solid solution nanoparticle catalysts for propane dehydrogenation (PDH) were prepared with four nominal Ga : Al atomic ratios (1 : 6, 1 : 3, 3 : 1, 1 : 0) using a colloidal synthesis approach. The structure, coordination environment and distribution of Ga and Al sites in these materials were investigated by X-ray diffraction, X-ray absorption spectroscopy (Ga K-edge) as well as 27Al and 71Ga solid state nuclear magnetic resonance. The surface acidity (Lewis or Brønsted) was probed using infrared spectroscopy with pyridine and 2,6-dimethylpyridine probe molecules, complemented by element-specific insights (Ga or Al) from dynamic nuclear polarization surface enhanced cross-polarization magic angle spinning 15N{27Al} and 15N{71Ga} J coupling mediated heteronuclear multiple quantum correlation NMR experiments using 15N-labelled pyridine as a probe molecule. The latter approach provides unique insights into the nature and relative strength of the surface acid sites as it allows to distinguish contributions from Al and Ga sites to the overall surface acidity of mixed (Ga,Al)2O3 oxides. Notably, we demonstrate that (Ga,Al)2O3 catalysts with a high Al content show a greater relative abundance of four-coordinated Ga sites and a greater relative fraction of weak/medium Ga-based surface Lewis acid sites, which correlates with superior propene selectivity, Ga-based activity, and stability in PDH (due to lower coking). In contrast, (Ga,Al)2O3 catalysts with a lower Al content feature a higher fraction of six-coordinated Ga sites, as well as more abundant Ga-based strong surface Lewis acid sites, which deactivate through coking. Overall, the results show that the relative abundance and strength of Ga-based surface Lewis acid sites can be tuned by optimizing the bulk Ga : Al atomic ratio, thus providing an effective measure for a rational control of the catalyst performance.

Multi-nuclear, high-pressure, operando FlowNMR spectroscopic study of Rh/PPh3 - catalysed hydroformylation of 1-hexene.

The hydroformylation of 1-hexene with 12 bar of 1 : 1 H2/CO in the presence of the catalytic system [Rh(acac)(CO)2]/PPh3 was successfully studied by real-time multinuclear high-resolution FlowNMR spectroscopy at 50 °C. Quantitative reaction progress curves that yield rates as well as chemo- and regioselectivities have been obtained with varying P/Rh loadings. Dissolved H2 can be monitored in solution to ensure true operando conditions without gas limitation. 31P{1H} and selective excitation 1H pulse sequences have been periodically interleaved with 1H FlowNMR measurements to detect Rh-phosphine intermediates during the catalysis. Stopped-flow experiments in combination with diffusion measurements and 2D heteronuclear correlation experiments showed the known tris-phosphine complex [RhH(CO)(PPh3)3] to generate rapidly exchanging isomers of the bis-phosphine complex [Rh(CO)2(PPh3)2] under CO pressure that directly enter the catalytic cycle. A new mono-phosphine acyl complex has been identified as an in-cycle reaction intermediate.

(Me2NH2)10[H2-Dodecatungstate] polymorphs: dodecatungstate cages embedded in a variable dimethylammonium cation + water of crystallization matrix.

Two polymorphs and a solvatomorph of a new dimethylammonium polytungstate—decakis(dimethylammonium) dihydrogendodecatungstate, (Me2NH2)10(W12O42)·nH2O (n = 10 or 11)—have been synthesized. Their structures were characterized by single-crystal X-ray diffraction and solid-phase NMR methods. The shape of the dodecatungstate anions is essentially the same in all three structures, their interaction with the cations and water of crystallization, however, is remarkably variable, because the latter forms different hydrogen-bonded networks, and provides a highly versatile matrix. Accordingly, the N–H⋯O and C–H⋯O hydrogen bonds are positioned in each crystal lattice in a variety of environments, characteristic to the structure, which can be distinguished by solid-state 1H-CRAMPS, 13C, 15N CP MAS and 1H–13C heteronuclear correlation NMR. Thermogravimetry of the solvatomorphs also reflect the difference and multiformity of the environment of the water molecules in the different crystal lattices. The major factors behind the variability of the matrix are the ability of ammonium cations to form two hydrogen bonds and the rigidity of the polyoxometalate anion cage. The positions of the oxygen atoms in the latter are favourable for the formation of bifurcated and trifurcated cation–anion hydrogen bonds, some which are so durable that they persist after the crystals are dissolved in water, forming ion associates even in dilute solutions. The H atom involved in furcated hydrogen bonds cannot be exchanged by deuterium when the compound is dissolved in D2O. An obvious consequence of the versatility of the matrix is the propensity of these compounds to form multiple polymorphs.

Carbonyl 13C-detect solution-state protein NMR experiments to circumvent amide-solvent exchange broadening: Application to β2-microglobulin

The 15N–1H heteronuclear single-quantum correlation (HSQC) technique in protein NMR spectroscopy suffers from line-broadening effects, such as chemical exchange of labile protons with solvent, and exchange broadening for residues undergoing conformational dynamics.The amide resonance of β2-microglobulin residue S88 is not observed in the HSQC spectrum but can be obtained through 13C-detect experiments that circumvent the problem of amide-solvent exchange broadening.Line broadening of S88 resonance beyond detection in the HSQC spectrum is not attributed to conformational exchange but rather to solvent exchange occurring on the order of ~103 s−1.

Activating Magnesium Electrolytes through Chemical Generation of Free Chloride and Removal of Trace Water.

Mg batteries are attractive next-generation energy storage systems due to their high natural abundance, inexpensive cost, and high theoretical capacity compared to conventional Li-ion based systems. The high energy density is achieved by electrodeposition and stripping of a Mg metal anode and requires the development of effective electrolytes enabled by a mechanistic understanding of the charge-transfer mechanism. The magnesium aluminum chloride complex (MACC) electrolyte is a good model system to study the mechanism as the solution phase speciation is known. Previously, we reported that minor addition of Mg(HMDS)2 to the MACC electrolyte causes significant improvement in the Mg deposition and stripping voltammetry resulting in good Coulombic efficiency on cycle one and, therefore, negating the need for electrochemical conditioning. To determine the cause of the improved electrochemistry, here we probe the speciation of the electrolyte after Mg(HMDS)2 addition using Raman spectroscopy, 27Al nuclear magnetic resonance spectroscopy, and 1H-29Si heteronuclear multiple bond correlation spectroscopy on MACC + Mg(HMDS)2 at various Mg(HMDS)2 concentrations. Mg(HMDS)2 scavenges trace H2O, but it also reacts with MACC complexes, namely, AlCl4-, to form free Cl-. We suggest that although both the removal of H2O and the formation of free Cl- improve electrochemistry by altering the speciation at the interface, the latter has a profound effect on electrodeposition and stripping of Mg.

Synthesis, NMR Characterization, and Antileukemic Activity of N-Nonanoylpiperazinyl-5α-Androstane-3α,17β-Diol A-Ring Derivatives

The combination of an androstane-3,17-diol nucleus and a 2β-N-alkylamidopiperazino sidechain is important for the anticancer activity of a new family of steroid derivatives. As the structure-activity relationship studies have so far been limited to the beta orientation of the substituent at position 2 of the steroid nucleus, a series of analogs (compounds 1–4) were synthesized to investigate the impact on biological activity of A-ring substitution. Nuclear magnetic resonance (NMR) analysis, especially using a series of 2D experiments, such as correlation spectroscopy (COSY), homonuclear Overhauser effect spectroscopy (NOESY), heteronuclear single-quantum correlation (HSQC), and heteronuclear multiple-bond correlation (HMBC) provided crucial information that was found essential in confirming the sidechain position and orientation of compounds 1–4. Assessment of their antiproliferative activity on leukemia HL-60 cells confirmed the best efficiency of the 2β-sidechain/3α-OH orientation (compound 1) compared to the other configurations tested (compounds 2–4).

Low-power synchronous helical pulse sequences for large anisotropic interactions in MAS NMR: Double-quantum excitation of 14N.

We develop a theoretical framework for a class of pulse sequences in the nuclear magnetic resonance (NMR) of rotating solids, which are applicable to nuclear spins with anisotropic interactions substantially larger than the spinning frequency, under conditions where the radiofrequency amplitude is smaller than or comparable to the spinning frequency. The treatment is based on average Hamiltonian theory and allows us to derive pulse sequences with well-defined relationships between the pulse parameters and spinning frequency for exciting specific coherences without the need for any detailed calculations. This framework is applied to the excitation of double-quantum spectra of 14N and is used both to evaluate the existing low-power pulse schemes and to predict the new ones, which we present here. It is shown that these sequences can be designed to be γ-encoded and therefore allow the acquisition of sideband-free spectra. It is also shown how these new double-quantum excitation sequences are incorporated into heteronuclear correlation NMR, such as 1H-14N dipolar double-quantum heteronuclear multiple-quantum correlation spectroscopy. The new experiments are evaluated both with numerical simulations and experiments on glycine and N-acetylvaline, which represent cases with "moderate" and "large" quadrupolar interactions, respectively. The analyzed pulse sequences perform well for the case of a "moderate" quadrupolar interaction, however poorly with a "large" quadrupolar interaction, for which future work on pulse sequence development is necessary.

Incorporated diffusion ordered heteronuclear multiple bond correlation spectroscopy, 3D iDOSY-HMBC. Merging of diffusion delay with long polarization transfer delay of HMBC

3D iDOSY-HMBC pulse sequences allow the simplification of HMBC data of mixtures via separation in the diffusion domain. The presented methods utilize incorporated DOSY approach, iDOSY, where the existing delays of the basic pulse sequence are utilized for diffusion attenuation. In the simplest form of the proposed 3D iDOSY-HMBC sequences, no extra delays or RF-pulses were required, only two diffusion gradients were added within HMBC polarization transfer delay.

Elucidation and Identification of an Antifungal Compound from Pseudomonas aeruginosa DA3.1 Isolated from Soil in Vietnam

Background: Fusarium sp. and Rhizoctonia sp. fungi have been always threats to short-term crops. In Vietnam, corn and soybean suffer serious losses annually. Therefore, it is necessary to utilize an environmentally friendly antifungal compound that is highly effective against phytopathogenic fungi. Pseudomonas sp. is a popular soil bacterial strain and well known for its high antifungal activity. Objectives: This study was carried out to evaluate and assess the antifungal activity of a local bacterial strain namely DA3.1 that was later identified as Pseudomonas aeruginosa. This would be strong scientific evidence to develop an environmentally friendly biocide from a local microorganism strain for commercial use. Methods: The antifungal compound was purified from ethyl acetate extraction of deproteinized cell culture broth by a silica gel column (CH2Cl2/MeOH (0% - 10% MeOH)). The purity of the isolated compound was determined by HPLC, and its molecular structure was elucidated using spectroscopic experiments including one-dimensional (1D) (1H NMR, 13C NMR, DEPT) and two-dimensional (2D) (HMBC and HSQC) spectra. The activity of the purified compound against Fusarium sp. and Rhizoctonia sp. fungi was measured using the PDA-disk diffusion method, and its growth-promoting ability was evaluated using the seed germination test of corn and soybean. Results: The results showed that the antifungal compound produced by Pseudomonas aeruginosa DA3.1 had a retention factor (Rf) of 0.86 on thin layer chromatography (TLC). Based on the evidence of spectral data including proton nuclear magnetic resonance (1H NMR), carbon nuclear magnetic resonance (13C NMR), distortionless enhancement by polarization transfer (DEPT), heteronuclear multiple bond correlation (HMBC), and heteronuclear single quantum coherence (HSQC), the chemical structure was elucidated as phenazine-1-carboxylic. The purified compound showed inhibitory activity against F. oxysporum and R. solani and exhibited the ability of the germination of corn and soybean seeds. The results revealed the benefit of native P. aeruginosa DA3.1 and phenazine-1-carboxylic acid for use as a biocontrol agent, as well as a plant growth promoter. Conclusions: The antifungal compound isolated from local Pseudomonas DA3.1 was identified as phenazine-1-carboxylic acid that posed high antifungal activity and was a plant germination booster.

Identifying the Unknown Content of an Ancient Egyptian Sealed Alabaster Vase from Kha and Merit’s Tomb Using Multiple Techniques and Multicomponent Sample Analysis in an Interdisciplinary Applied Chemistry Course

This article highlights the multianalytical study of exuded liquid from an ancient Egyptian sealed alabaster vase by Master’s students in an applied chemistry for cultural heritage course. Master students are introduced to the field of Archaeometry that see the collaboration of experts in different areas of research such as conservators, curators of museums, physicists, chemists, etc. The sample is a residue exuded on the linen strip sealing an ancient Egyptian alabaster vase (inventory number S.8448) from the collection of the Museo Egizio in Turin (Italy). The students start to plan the noninvasive investigation by X-ray fluorescence (XRF), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDS) for the inorganic compounds characterization, followed by the extraction of the organic components (such as oils, fats, and waxes) to be analyzed by high-resolution 1H and 13C nuclear magnetic resonance (NMR) spectroscopy and 2D NMR correlation spectroscopy (COSY), heteronuclear single quantum coherence (HSQC), and long-range heteronuclear correlation (HMBC) techniques and by gas chromatography coupled with mass spectrometry (GC/MS). Reference standards, spectral databases, and published data on similar artifacts served as the basis for the interpretation of the instrumental results. The approach was introduced in the course of Applied Chemistry for Cultural Heritage for Master students in Archaeology (University of Palermo, Italy), where the need is to know how to approach the scientific investigation together with the conservation scientists and how to manage with a very low amount of sample. Pedagogically, the approach introduces students to the main techniques currently used in the field of Archaeometry while reinforcing fundamental concepts in sample collecting and multicomponent microsample analysis. This interdisciplinary approach provides a unique experience that demonstrates chemistry’s broad applicability outside of the traditional laboratory. Students are guided to identify the inorganic and organic components of the exudate liquid: the first one is ascribable to clay minerals iron oxides, which could impart the brown color to the sample; the second one is ascribed to triglycerides of various kinds, which probably comes from vegetable oil.