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Research

2D NMR: Overview of Heteronuclear Correlation Techniques

Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds&#39; structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other axis. Cross-peaks reveal the connection between specific protons and X-nuclei.
The main heteronuclear correlation experiments include HETCOR, HSQC, HMQC, and HMBC. HSQC, HMQC, and HMBC record the proton spectrum, whereas the X-nucleus spectrum is recorded by standard heteronuclear correlation spectroscopy or HETCOR. The HETCOR-HMQC-HSQC group of experiments cannot detect X-nuclei without an attached proton. HETCOR is used when ultrahigh peak resolution is required along the X-nucleus axis.
The heteronuclear single quantum correlation technique, or HSQC, investigates proton-to-X-nucleus single bond correlations. In an HSQC experiment, polarization is transferred from a proton nucleus to a neighboring X-nucleus and then back to the proton nucleus. The signal from the proton nuclei is then recorded. HSQC generates a spectrum like that of heteronuclear multiple-quantum correlation spectroscopy or HMQC but uses a different suppression method, providing better X-nucleus resolution along the axis.
Heteronuclear multiple-bond correlation, or HMBC, is a proton-detected experiment, like HMQC but with a longer initial delay time. This experiment is used to observe long-range proton to X-nucleus connectivity separated by 2-3 bonds, with some experiments going out to 4 or 5 bonds. In an HMBC experiment, direct one-bond correlations are suppressed as part of the sequence. The spectra of HMBC give rise to cross peaks that correlate which protons are coupled to what other specific X-nucleus is more than one bond away. The gradient HMBC (gHMBC) pulse sequence offers better suppression of interfering signals, making it particularly useful for analyzing complex molecules with overlapping signals.

Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an ...

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does not have any diagonal peaks. All the cross-peaks indicate the protons directly attached to each carbon in a molecule. Similar to the COSY spectrum, imaginary lines are drawn in horizontal and vertical directions to identify the correlated peaks.
For example, the HSQC spectrum of 2-chlorobutane has a carbon signal at 11 ppm that can be assigned to the shielded methyl group and correlated to the proton multiplet at 1.01 ppm. The carbon signal at 24.8 ppm arises from the deshielded methyl group, which correlates to the proton multiplet at 1.52 ppm. Similarly, the methylene carbon signal at 33.3 ppm and methine carbon signal at 60.4 ppm correlate to the multiplets at 1.72 and 3.96 ppm, respectively.&#160; HSQC is a very useful technique for structurally elucidating complex molecules and is routinely used in protein NMR studies.

Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a ...

Double Resonance Techniques: Overview

Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
Spin decoupling is usually achieved by irradiating the sample atoms with a suitable radio frequency (rf) pulse sequence, effectively eliminating all the coupling with one nuclide. This simplifies the observed spectrum, making it easier to analyze and understand the relationships between different nuclei.
Double resonance experiments can be classified as heteronuclear or homonuclear, depending on whether the two sets of nuclei are of different isotopes or the same isotope, respectively. They can also be selective or nonselective, depending on whether the irradiating frequency covers only a portion or all of the resonance frequencies.
In nonselective heteronuclear decoupling, samples are exposed to an appropriate radiofrequency range to remove all coupling with one nuclide. However, increased external field strengths necessitate more powerful irradiation across broader frequency ranges. In certain instruments, continuous irradiation creates enough heat to cause damage to thermally sensitive samples.
To overcome these problems, modern methods using a series of pulses and exact timing delays, like J-modulated spin echo pulse sequences, can be employed to eliminate or adjust coupling effects within the spectrum.
For example, the attached proton transfer (APT) experiment applies a J-modulated spin echo pulse sequence to focus on the phase of detected carbon signals. Carbon atoms attached to an even number of protons exhibit positive signals in the spectrum, while those bonded to an odd number of protons appear as negative signals.
It utilizes a combination of a 180&#176; proton pulse and broadband decoupling to simplify spectral interpretation and assign multiplicities. The 180&#176; pulse is applied to the protons during the APT experiment. Its role is to refocus the spin coupling evolution that naturally occurs due to interactions between protons and carbons.
Specifically, after the 90&#176; pulse creates transverse magnetization, the 180&#176; pulse ensures that the effects of coupling are frozen during the initial period where the decoupler is turned off. This means the chemical shifts evolve only during the time 1/J&#8203; (the coupling constant period) after the first 180&#176; pulse.
By carefully timing this pulse sequence, the experiment isolates carbon multiplicities by utilizing the J-coupling between carbons and their attached protons. After the coupling evolution period (1/J&#8203;), broadband decoupling is turned on. This removes the J-coupling signals, collapsing multiplets into single peaks for each carbon resonance.
The decoupling ensures that carbon signals are free of splitting, allowing for a clear differentiation of signals based on phase (positive or negative) rather than splitting patterns.
The APT technique selectively enhances carbon signals, providing valuable information about carbon-proton connectivity, the number of attached hydrogen atoms, and the overall molecular structure.

Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or ...

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) is an advanced Nuclear Magnetic Resonance (NMR) technique specifically designed to detect and enhance the signals of low-abundance nuclei, such as carbon-13 and nitrogen-15, in small molecules. The fundamental principle behind INEPT is the transfer of polarization from a more abundant and highly polarizable nucleus, typically hydrogen-1, to the low-abundance nucleus of interest. This process effectively boosts the NMR signal of the low-abundance nuclei, making them more easily detectable.
The INEPT technique involves a series of carefully timed radiofrequency (RF) pulses applied to the sample. These RF pulses excite the more abundant nuclei, causing them to align their spins in a specific manner. Through the process of spin coupling, the polarization of the more abundant nuclei is then transferred to the low-abundance nuclei. As a result, the NMR signal of these low-abundance nuclei is significantly enhanced, allowing for a more detailed analysis of the molecular structure.
Various pulse sequences have been developed to improve the observability of these low-abundance nuclei when coupled to a high-receptivity nucleus. The INEPT pulse sequence combines a spin echo and selective population inversion, which includes 90&#176; and 180&#176; pulses on the high-sensitivity (the I spins) and low-sensitivity nuclei (the S spins), respectively. Coupling-constant-based delays between pulses facilitate magnetization transfer through space or bond interactions. Additional RF pulses and delays invert the low-gamma magnetization, producing an observable NMR signal. The net result is an enhanced NMR signal from low-gamma nuclei that would otherwise be challenging to detect.
Various pulse sequences have been developed to improve the detection of low-abundance nuclei when coupled with high-receptivity nuclei such as protons. The INEPT sequence yields enhanced signals for insensitive nuclei, with half of the CH group producing negative peaks and the other half positive peaks. This pattern allows for discrimination between different types of carbon-hydrogen bonds within the molecule.
Decoupling methods can be employed to enhance sensitivity and resolution in distinguishing various carbon-hydrogen bonds, making INEPT an invaluable tool in the chemical and pharmaceutical industries.

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) is an advanced Nuclear Magnetic Resonance (NMR) technique specifically designed to detect and ...

Two-Dimensional (2D) NMR: Overview

The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse. The second step is the evolution or wait period t1, which is incremented systematically, and no data is observed during this period. The next step is the mixing period, in which a second radiofrequency pulse is introduced, which transfers the magnetization to the spins of nucleus B. In the last step, the magnetization of nuclear spins from B is detected during the acquisition time t2 for each t1. Time domains t1 and t2 are Fourier transformed into frequency domains f1 and f2, and these correspond to two dimensions of 2D spectra, while the third dimension is the intensity. In 2D experiments, coupling between the same kind of nucleus is called homonuclear 2D experiments, whereas coupling between two types of nucleus is called heteronuclear 2D experiments. 2D spectra can be displayed as stacked or contour plots. When presented as stacked plots, 2D spectra are difficult to interpret. The contour plot is the horizontal cross-section of a stacked plot where the size of the circles shows the signal intensity. Its interpretation is straightforward and it is preferred for data analysis.

The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily ...

2D NMR: Overview of Homonuclear Correlation Techniques

Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that utilizes a second 90-degree pulse to achieve high sensitivity and resolution. It excels in analyzing small to medium-sized molecules and provides information on directly coupled protons. COSY45, similar to COSY90, uses 45-degree pulses instead of 90-degree pulses. This results in reduced sensitivity but improved resolution.
Long-range COSY (LR-COSY) extends the COSY experiment to detect scalar couplings between nuclei further apart in the molecule. It achieves this by combining multiple pulse sequences and magnetization transfers.
Relayed COSY is a variant of COSY that adds extra pulses to the standard pulse sequence to produce cross-peaks between cross-peaks, showing specific molecular connections hidden by overlapping signals.
TOCSY (Total Correlation Spectroscopy), also known as HOHAHA (Homonuclear Hartmann-Hahn Spectroscopy), reveals all through-bond connectivity between all coupled nuclei. TOCSY is especially useful for analyzing molecules with overlapping signals and complex coupling patterns.
NOESY (Nuclear Overhauser Effect Spectroscopy) measures the nuclear overhauser effect, which arises from dipole-dipole interactions between nuclei. It provides information about spatial proximity and can be used to determine the three-dimensional structure of a molecule.
ROESY (Rotating-frame Overhauser Enhancement Spectroscopy) is similar to NOESY but employs a different mechanism to obtain through-space correlations. It provides information about spatial proximity and is particularly useful for studying molecules with weak NOE signals.
In conclusion, choosing a COSY variant depends on the molecule or reaction under investigation and the desired information. The right COSY variant can be determined based on molecular size, coupling complexity, and the need for spatial or long-range connectivity data.

Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations ...

2D NMR: Homonuclear Correlation Spectroscopy (COSY)

Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY spectrum is symmetric about the diagonal, with signals above and below the diagonal called cross-peaks or off-diagonal peaks. An imaginary line is drawn from diagonal peaks parallel to the horizontal axis, leading to a cross peak. A vertical line is then drawn from the same cross peak towards the diagonal peak to correlate the peaks. The off-diagonal peaks correlating the multiplets at 1.25 and 4.2 ppm indicate that the methyl protons are coupled to the methylene protons. The signal at 2 ppm is not associated with cross-peaks because the protons are not involved in homonuclear coupling. A COSY spectrum is useful when overlapping multiplets complicate the corresponding 1D spectrum.

Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal ...

&sup1;H NMR of Labile Protons:  Deuterium (&sup2;H) Substitution

This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
<img alt="Figure1" src="/files/ftp_upload/12280/12280-PT_Figure_1.svg" style="text-align: center; height: 700px;" />
Figure 1. The comparison of the proton NMR spectra of pure dry ethanol and the same in D2O solvent.
The signals from deuterium are not observed in a proton NMR spectrum, as deuterium&#8211;proton coupling is negligible, and no spin-spin splitting occurs between them. As shown in Figure 1, the NMR spectra of pure ethanol and ethanol in deuterium, the disappearance of the triplet in the latter, confirms that it corresponds to the labile &#8211;OH proton. Furthermore, the methylene protons that initially showed up as a multiplet transform to a quartet, proving that the multiplet is due to the coupling with the &#8211;OH proton and the methyl protons.

This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is ...

&sup1;H NMR of Labile Protons: Temporal Resolution

Protons bonded to heteroatoms such as nitrogen and oxygen exhibit a range of chemical shift values. This is due to the varying degree of hydrogen bonding between the proton and the heteroatom in other molecules. The extent of hydrogen bonding affects the electron density around the proton, thereby giving different chemical shift values for the protons in the proton NMR spectrum.
The &#8211;OH proton in alcohols typically appears in the range of &#948; 2 to 5 ppm but can vary depending on the specific alcohol and conditions. This &#8211;OH proton undergoes rapid proton exchange with other molecules and is referred to as a labile proton. So, the exact &#948; value of the &#8211;OH proton and the spin-spin coupling involving the &#8211;OH proton and the other protons in the alcohol depends on the rate of the &#8211;OH proton exchange. The proton exchange rate, in turn, depends on multiple factors, including the purity of the alcohol sample, temperature, and solvent.
<img alt="Figure1" src="/files/ftp_upload/12279/12279-PT_Figure_1.svg" style="text-align: center; height: 350px;" />
Figure 1. The triplet of a &#8211;OH proton in the proton NMR spectrum of neat ethanol.
For example, the &#8211;OH proton exchange in a pure dry ethanol sample is very slow on the NMR time scale, giving enough time for the spin-spin coupling. Consequently, as depicted in Figure 1, the proton NMR spectrum of pure dry ethanol shows a triplet for the &#8211;OH proton due to the spin-spin coupling with adjacent methylene protons. The methylene protons are seen as a multiplet due to the coupling with the &#8211;OH proton and the methyl protons.
The presence of trace amounts of an acid or base increases the proton exchange rate. The proton exchange becomes rapid, and the NMR spectrometer only records an average of all the possible environments for the &#8211;OH proton.
<img alt="Figure2" src="/files/ftp_upload/12279/12279-PT_Figure_2.svg" style="text-align: center; height: 325px;" />
Figure 2. The broad singlet of a &#8211;OH proton in the proton NMR spectrum of neat ethanol.
As a result, as recorded in Figure 2, the &#8211;OH proton exhibits a broad singlet. The protons from the adjacent methylene group present a quartet where the splitting is caused only by the adjoining methyl group.

Protons bonded to heteroatoms such as nitrogen and oxygen exhibit a range of chemical shift values. This is due to the varying degree of hydrogen bonding ...

Nuclear Overhauser Enhancement (NOE)

Irradiation of a spin-active nucleus causes an increase or decrease in the signal intensity of neighboring nuclei that are not necessarily chemically bonded or involved in J-coupling. &#160;This phenomenon, called the Nuclear Overhauser Enhancement (NOE), results from through-space interactions between the nuclear spins. The NOE effect decreases with increasing internuclear distance and is generally not observed beyond 4 angstroms. In NOE, dipole-dipole interactions between neighboring spin-active nuclei result in the transfer of nuclear spin polarization. These interactions can cause an increase or decrease in the signal intensity, which results in a positive or negative NOE, respectively.
NOE can be applied to improve the signal intensity of less sensitive nuclei such as carbon-13. Proton-decoupled carbon-13 spectra show a positive NOE effect, where the carbon signal intensities are substantially increased compared to those in proton-coupled spectra.
Targeted irradiation of specific nuclei can also reveal through-space proximity, which helps confirm stereochemistry and verify spectral peak assignments. The NOE effect is also applied in 2D NMR experiments, such as Nuclear Overhauser Effect Spectroscopy (NOESY), to determine the 3D structure of proteins.

Irradiation of a spin-active nucleus causes an increase or decrease in the signal intensity of neighboring nuclei that are not necessarily chemically ...

&sup1;H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR

The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
<img alt="Figure1" src="/files/ftp_upload/13016/13016-PT_Figure_1.svg" style="text-align: center; height: 480px;" />
Figure 1. The temperature-dependent proton NMR spectra of cyclohexane.
Figure 1 depicts the proton NMR spectrum of the deuterium substituted sample, recorded at various temperatures. At room temperature, the rapid ring flipping of cyclohexane results in a single sharp peak. As the temperature is lowered to &#8722;60&#176;C, the rate of chair-chair interconversion reduces, resulting in the broadening of the peak.
Further lowering of the temperature broadens the peak, forming a saddle that splits into two peaks. On reaching a temperature lower than &#8722;89&#176;C, two sharp, well-resolved peaks corresponding to the equatorial and axial protons are formed. The peaks corresponding to the equatorial and axial protons occur at &#948; 1.62 and 1.14, respectively. At such low temperatures, the rate of chair-chair interconversion is reduced significantly, enabling the detection of the two distinct sets of protons on the NMR time scale.

The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one ...

&sup1;H NMR of Conformationally Flexible Molecules: Temporal Resolution

At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the axial and equatorial protons, resulting in a single peak at &#948; 1.4.
Similarly, NMR averages all the conformations of any molecule with rapid conformational equilibria. For example, the CH3 protons in bromoethane have a single resonance and coupling constant for splitting by the CH2 protons due to the rapid internal rotation of the carbon-carbon bond. Lastly, besides conformational equilibria, the time-averaging effect of NMR spectroscopy is also observed in some chemical reactions.

At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately ...

Chemical Ionization (CI) Mass Spectrometry

The molecular ion peak of a molecule in the mass spectrum provides vital information for molecular identification. However, conventional electron impact ionization can lead to the rapid dissociation of some molecular ions before they reach the detector. A milder ionization method is required to increase the lifetime of such ionized analyte molecules. Chemical ionization (CI) is a gas-phase protonation reaction useful for mass-analyzing analyte molecules that are easily protonated to yield the corresponding conjugate acid. In this process, the sample is mixed with an excess reagent gas, which ensures the electron impact occurs primarily on the reagent gas. The charged species formed from the reagent protonates the analyte molecule, producing a relatively stable protonated analyte (conjugate acid) compared to the molecular ion. This results in an M+1 peak in the mass spectrum. The conjugate acid may then undergo fragmentation that generates additional signals.
For example, the CI of di-sec-butyl ether in methane gas illustrates this process. When the ether is mixed with excess methane gas as a reagent, electron impact occurs on methane rather than the ether. The resulting methane radical cation can react with another methane molecule to generate a methane radical and a methanium ion. The methanium ion is a source of gas-phase protons, which can protonate the ether to form its conjugate acid. The sequential reactions during the chemical ionization of di-sec-butyl ether are depicted in Figure 1.
<img alt="Figure1" src="/files/ftp_upload/12300/12300_PT_Figure_1.svg" style="text-align: center; height: 175px;" />
Figure 1: Chemical ionization of di-sec-butyl ether methane mixture.
This conjugate acid of the ether (m/z = 131) is relatively more stable than the molecular ion (m/z = 130) of ether formed via the conventional electron impact ionization. In the traditional route, the molecular ion undergoes fragmentation via &#945; cleavage, giving a signal at m/z = 101. Figure 2 illustrates the reactions that occur during electron impact ionization directly on di-sec-butyl ether.
<img alt="Figure2" src="/files/ftp_upload/12300/12300_PT_Figure_2.svg" style="text-align: center; height: 175px;" />
Figure 2: Electron impact ionization of di-sec-butyl ether and fragmentation of the molecular ion.
So, the mass spectra of di-sec-butyl ether ionized via CI feature an M+1 peak at m/z = 131. On the other hand, mass spectra of di-sec-butyl ether ionized via electron impact ionization do not show any peak at the m/z value of its molecular weight. Figures 3a and 3b show the mass spectra of di-sec-butyl ether ionized via the electron impact and chemical ionization methods, respectively.
<img alt="Figure3" src="/files/ftp_upload/12300/12300_PT_Figure_3.svg" style="text-align: center; height: 225px;" />
Figure 3:&#160;a) No m/z = 130 peak is visible in the mass spectrum obtained via electron impact ionization of di-sec-butyl ether. b) The m/z = 131 peak is clearly visible in the mass spectrum obtained via chemical ionization of di-sec-butyl ether.

The molecular ion peak of a molecule in the mass spectrum provides vital information for molecular identification. However, conventional electron impact ...

Electrospray Ionization (ESI) Mass Spectrometry

Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the gaseous phase. The analyte biomolecule is premixed with an ionic liquid&#8212;a nonmolecular compound serving as the ionization source. The solution is sprayed through a high-voltage capillary to generate a fine aerosol. The voltage difference applied between the capillary and the entrance of the analysis chamber causes the movement of charged droplets toward the analysis chamber. The ionic liquid from the aerosol evaporates during this movement, leaving behind a variably protonated or ionized analyte molecule. These charged species are detected in the mass spectrum at an m/z value equivalent to the sum of the analyte molecule&#39;s molecular mass and the ions&#39; molecular weight from the ionic liquid associated with the analyte molecule.
The position of mass signals varies depending on the type of cations present in the ionic liquid. For example, the presence of sodium ions in the ionic liquid leads to an (M+23) peak in the mass spectra, while the presence of a proton results in an (M+1) peak.

Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional ...

Mass Spectrometry: Aldehyde and Ketone Fragmentation

In mass spectrometry, the fragmentation of aliphatic aldehydes and ketones generally occurs through three key mechanisms: &#945;-cleavage, inductive cleavage, and the McLafferty rearrangement.
<ol>
	<li>&#945;-Cleavage 
	&#945;-Cleavage is a common fragmentation in carbonyl compounds where the bond adjacent to the carbonyl group is broken. This process produces an alkyl radical and an acylium cation, which is often detected in the mass spectrum due to its stability.</li>
	<li>Inductive Cleavage 
	In inductive cleavage, fragmentation yields an acyl radical and an alkyl cation. This occurs when electrons are drawn toward the carbonyl, weakening adjacent bonds and resulting in a distinctive fragmentation pattern. For example, in the mass spectrum of 5-methyl-2-hexanone, inductive cleavage produces an alkyl cation at a mass-to-charge (m/z) ratio of 71.</li>
	<li>McLafferty Rearrangement 
	The McLafferty rearrangement is a specific fragmentation pattern for carbonyl compounds with &#947;-hydrogen (hydrogen on the third carbon from the carbonyl group). The molecule undergoes a six-membered cyclic transition in this rearrangement, yielding a radical cation and a neutral alkene. For 5-methyl-2-hexanone, this rearrangement results in a peak at m/z 58.</li>
</ol>
Example: Fragmentation of 5-methyl-2-hexanone 
In the mass spectrum of 5-methyl-2-hexanone, the molecular ion undergoes these fragmentation patterns:
<ol style="list-style-type: circle;">
	<li>Inductive cleavage produces an alkyl cation at m/z 71.</li>
	<li>&#945;-Cleavage results in the base peak at m/z 43.</li>
	<li>McLafferty rearrangement creates a distinct peak at m/z 58.</li>
</ol>
An additional diagnostic feature in aldehydes is the M&#8722;1 peak, which arises due to the &#945;-cleavage of the aldehyde proton. In mass spectrometric analysis, this peak can help identify aldehydes, specifically among other carbonyl compounds.
Understanding these fragmentation pathways is crucial for interpreting mass spectra, particularly in identifying functional groups and distinguishing between similar compounds.

In mass spectrometry, the fragmentation of aliphatic aldehydes and ketones generally occurs through three key mechanisms: &#945;-cleavage, inductive ...

Mass Spectrometry: Alkyl Halide Fragmentation

Chlorine isotopes exist as 35Cl and 37Cl in a 3:1 ratio, while bromine isotopes exist as 79Br and 81Br in a 1:1 ratio. The mass spectrum of alkyl halides typically produces two distinct molecular ion peaks, the molecular ion peak, [M], and the molecular ion plus two, [M + 2] peak. The relative heights of these two peaks are proportional to the isotopic abundance ratios of the halide. For example, 2&#8208;chloropropane and 1&#8208;bromopropane display two peaks with relative peak heights in a 3:1 and 1:1 ratio, respectively.
Alkyl halides typically lose a non-bonding electron from the halogen to form molecular ions. The base peak results from the heterolytic cleavage of the carbon&#8211;halogen bond, where the bonding electrons go to the most electronegative atom, forming a cation and halogen atom. Interestingly, the &#9082;-cleavage is observed only in alkyl chlorides, not alkyl bromides. In alkyl chlorides, the&#160; C&#8211;Cl and C&#8211;C bonds exhibit similar strengths, while in alkyl bromides, the C&#8211;Br bond is weaker than the C&#8211;C bond. Consequently, the alkyl chlorides form a stable cation, where the positive charge is shared between the two atoms.
While less commonly discussed, fluoroalkanes and iodoalkanes exhibit unique fragmentation behavior. Fluoroalkanes, due to the strength of the C&#8211;F bond and the presence of only one isotope (19F), show limited fragmentation and lack isotopic peaks. Iodoalkanes, with their weak C&#8211;I bond and a single isotope (127I), predominantly fragment by losing iodine, resulting in simpler spectra with less isotopic splitting.

Chlorine isotopes exist as 35Cl and 37Cl in a 3:1 ratio, while bromine isotopes exist as 79Br and 81Br in a 1:1 ratio. The mass spectrum of alkyl halides ...

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and characterization of various biomolecules, including proteins, peptides, nucleic acids, and carbohydrates. MALDI spectrometry is widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.
The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix material. The matrix is usually an organic compound, such as a small organic acid or a crystalline substance, that facilitates ionization. This analyte-matrix mixture is then deposited onto a target surface, inserted into the mass spectrometer, and dried. During drying, the matrix forms small crystals that incorporate the analyte molecules within their structure.
Once the sample is prepared, a pulsed laser is directed onto the target surface. The matrix molecules absorb the laser beam energy, causing them to heat up and vaporize rapidly. This process is known as laser desorption. As the matrix molecules vaporize, they carry the embedded analyte molecules. The vaporized analyte molecules become ionized through a process called proton transfer. The proton transfer occurs between the matrix and the analyte molecule. The matrix molecule donates a proton to the analyte molecule, resulting in the formation of protonated ions. The analyte ions are then accelerated by an electric field and further separated based on their mass-to-charge ratio in a time-of-flight mass analyzer.
The mass spectrum obtained from MALDI spectrometry provides valuable information about the mass and relative abundance of the analyzed biomolecules.

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and ...

Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation

The fragmentation patterns observed for compounds such as carboxylic acids, esters, and amides in the mass spectra include &#9082;-cleavage and McLafferty rearrangement. Fragmentation by &#9082;-cleavage preferentially occurs at the carbon-carbon bond at the &#9082;-position next to the carboxylic group to generate a neutral radical and a cation. Long chain compounds with hydrogen at their &#947;-carbon undergo McLafferty rearrangement to give a radical cation and a neutral alkene.
For example, the fragmentation of butyramide is demonstrated in Figure 1. The molecular ion peak of butyramide, which contains one nitrogen atom, shows an odd mass-to-charge ratio of 87. Fragmentation of butyramide molecular ion by &#9082;-cleavage produces a cation at a mass-to-charge ratio 44. In addition to &#9082;-cleavage, butyramide molecular ion also undergoes fragmentation via McLafferty rearrangement. The base peak at the mass-to-charge ratio 59 arises due to McLafferty rearrangement.
<img alt="Figure1" src="/files/ftp_upload/13051/13051_PT_Figure_1.svg" style="text-align: center; height: 250px;" />

The fragmentation patterns observed for compounds such as carboxylic acids, esters, and amides in the mass spectra include &#9082;-cleavage and McLafferty ...

Mass Spectrometry: Aromatic Compound Fragmentation

Upon ionization, aromatic compounds generate a molecular ion that is observed as a prominent peak in their mass spectra. For example, the molecular ion peak for benzene appears at a mass-to-charge ratio of 78, while toluene is observed at a mass-to-charge ratio of 92. The molecular ion benzene is highly stable and does not readily undergo further fragmentation due to the significant amount of energy required to disrupt the aromatic stability of the benzene ring. In contrast, the molecular ion of alkyl&#8208;substituted benzenes, such as toluene, can undergo fragmentation by losing a hydrogen atom, forming a benzylic carbocation. This carbocation can rearrange to form the resonance-stabilized tropylium cation, which appears in the mass spectra as a strong peak. In the case of toluene, this strong peak is observed at a mass-to-charge ratio of 91.

Upon ionization, aromatic compounds generate a molecular ion that is observed as a prominent peak in their mass spectra. For example, the molecular ion ...

Mass Spectrometry: Alcohol Fragmentation

Alcohols (R-OH) ionize to lose one non-bonded electron from the oxygen atom, forming molecular ions. Due to their tendency to fragment rapidly, the intensity of the molecular ion peak in the mass spectrum is weak or sometimes absent. The fragmentation patterns for alcohols occur in two ways, i.e. &#9082;-cleavage and dehydration. During &#9082;-cleavage, the bond at the &#9082;-position adjacent to the hydroxyl group cleaves to give a resonance-stabilized cation and a radical. However, intramolecular dehydration involves losing a water molecule from an alcohol, forming an alkene cation, which shows a peak at M&#8722;18. Consider the fragmentation of butanol as shown below. &#9082;-cleavage produces a base peak at a mass-to-charge ratio of 31. In the case of dehydration, the fragmentation forms an alkene radical cation at a mass-to-charge ratio of 56.
<img alt="Figure1" src="/files/ftp_upload/13046/13046-PT_Figure_1.svg" style="text-align: center; height: 175px;" />

Alcohols (R-OH) ionize to lose one non-bonded electron from the oxygen atom, forming molecular ions. Due to their tendency to fragment rapidly, the ...

Mass Spectrometry: Amine Fragmentation

Amines can be identified using mass spectroscopy based on their characteristic fragmentation patterns. The molecular ions of amines undergo fragmentation via &#9082;-cleavage. The &#9082;-cleavage of the carbon-carbon bonds in amines generates an alkyl radical and resonance-stabilized nitrogen-containing cation.
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing a single or an odd number of nitrogen atoms produces a molecular ion with an odd molecular weight, while a compound with no or an even number of nitrogen atoms generates a molecular ion with an even molecular weight.
For example, the mass spectrum of 3-methyl-1-butanamine, which contains one nitrogen atom, displays a molecular ion having an odd mass-to-charge ratio of 87. Fragmentation of this molecular ion via &#9082;-cleavage produces a base peak, which features a mass signal at a mass-to-charge ratio of 30, corresponding with the loss of the 2-methyl propyl radical.
Similarly, the fragmentation of triethylamine via &#9082;-cleavage generates a cation at a mass-to-charge ratio of 86. The molecular ion peak shows an odd mass-to-charge ratio of 101, as shown below.
<img alt="Figure1" src="/files/ftp_upload/13048/13048_PT_Figure_1.svg" style="text-align: center; height: 105px;" />

Amines can be identified using mass spectroscopy based on their characteristic fragmentation patterns. The molecular ions of amines undergo fragmentation ...

Mass Spectrometry: Cycloalkene Fragmentation

The molecular ions of cycloalkenes undergo fragmentation via a retro-Diels&#8211;Alder reaction.

The reaction proceeds via the cleavage of two carbon-carbon bonds in the cycloalkene to yield ethene. The remaining part of the cycloalkene structure is a dienyl radical cation with a molecular weight of 28 u lower than the molecular ion. This fragmentation pathway is similar to the fragmentation of cycloalkanes releasing ethene, differing only in the resultant radical cation. An alkyl species forms from the cycloalkane, whereas a dienyl species originates from the cycloalkene.

Retro-Diels&#8211;Alder reactions of cycloalkenes are also similar to the McLafferty-rearrangement-type fragmentation observed in acyclic alkenes with hydrogen on the &#947; carbon. In both cases, cleavage occurs at two positions, yielding an ethene molecule and an alkenyl species. In acyclic alkenes, the cleavage occurs at one carbon-carbon bond and a carbon-hydrogen bond, along with rearranging hydrogen from one carbon to another. In cyclic alkenes, cleavage occurs at two carbon&#8211;carbon bonds without any atomic rearrangement. Figure 1 depicts the fragmentation of cyclohexene via a retro-Diels&#8211;Alder reaction.
<img alt="Figure1" src="/files/ftp_upload/13044/13044-PT_Figure_1.svg" style="text-align: center; height: 90px;" />
Figure 1: Fragmentation of cyclohexene via a retro-Diels&#8211;Alder reaction.
In addition to the retro-Diels&#8211;Alder reaction, branched cycloalkenes exhibit fragmentation via cleavage of the side chain to form a cycloalkene cation. For example, the fragmentation of 1-methyl-1-cyclohexene via a retro-Diels&#8211;Alder reaction and side-chain cleavage are shown in Figure 2.
<img alt="Figure2" src="/files/ftp_upload/13044/13044-PT_Figure_2.svg" style="text-align: center; height: 225px;" />
Figure 2: Fragmentation of 1-methyl-1-cyclohexene via (top) a retro-Diels&#8211;Alder reaction; (bottom) side-chain cleavage.
The corresponding signals of the isoprenyl radical cation and the cyclohexenyl cation are evident in the mass spectrum of 1-methyl-1-cyclohexene shown in Figure 3.
<img alt="Figure3" src="/files/ftp_upload/13044/13044-PT_Figure_3.svg" style="text-align: center; height: 250px;" />
Figure 3: Mass spectrum of 1-methyl-1-cyclohexene.

The molecular ions of cycloalkenes undergo fragmentation via a retro-Diels&#8211;Alder reaction.

The reaction proceeds via the cleavage of two ...

Mass Spectrometry: Alkyne Fragmentation

The fragmentation of alkynes preferentially occurs at the carbon&#8211;carbon bond between the &#945; and &#946; carbon of the alkyne bond to generate a 3-propynyl cation (or propargyl cation). In terminal alkynes, there is the only type of fragmentation that yields the 3-propynyl cation. The unsubstituted 3-propynyl cation exhibits a peak at a mass-to-charge ratio of 39. In internal alkynes, the 3-propynyl cation is substituted. For example, 2-pentyne fragments into methyl-substituted 3-propynyl cation, which subsequently features as a mass signal at the mass-to-charge ratio of 56. The 3-propynyl cation is resonance-stabilized, and the corresponding mass signal is intense in the mass spectrum of alkynes. Figure 1 shows the fragmentation of 1-pentyne into an ethyl radical and a resonance stabilized 3-propynyl cation.
<img alt="Figure1" src="/files/ftp_upload/13045/13045-PT_Figure_1.svg" style="text-align: center; height: 115px;" />
Figure 1: Fragmentation of 1-pentyne molecular ion to yield the 3-propynyl cation.
Another fragmentation pathway observed in terminal alkynes is the loss of hydrogen from the terminal sp carbon. This results in a mass signal with a mass-to-charge ratio of one unit less than the molecular ion. This M&#8211;1 peak is very intense and becomes the base peak in the mass spectra of many alkynes, so it can be used to identify alkynes.
<img alt="Figure2" src="/files/ftp_upload/13045/13045-PT_Figure_2.svg" style="text-align: center; height: 275px;" />
Figure 2: The mass spectrum of 1-pentyne.

The fragmentation of alkynes preferentially occurs at the carbon&#8211;carbon bond between the &#945; and &#946; carbon of the alkyne bond to generate a ...

Mass Spectrometry: Branched Alkane Fragmentation

This lesson delves into the mass spectrometry of branched alkane fragmentation. Branched alkanes possess secondary or tertiary carbon atoms, which generate relatively stable carbocations if the cleavage occurs at the branching point. The high stability of carbocations drives the instant fragmentation of branched alkanes. Accordingly, the branched alkane&#39;s molecular ion peak is very weak or invisible in the mass spectra, especially in comparison to a linear alkane.
<img alt="Figure1" src="/files/ftp_upload/13041/13041-PT_Figure_1.svg" style="text-align: center; height: 275px;" />
Figure 1. Fragmentation pathway of 2-methylbutane (top), neopentane (middle), and n-pentane (bottom) molecular ions.
Figure 1 shows the most feasible fragmentation pathway observed in 2-methyl butane, neopentane, and n-pentane molecular ions. 2-methyl butane and neopentane fragment to yield secondary and tertiary carbocations, respectively. The stability of these carbocations drives the fragmentation reaction, even though the co-produced methyl radical is relatively unstable. In contrast, the cleavage of n-pentane leading to the methyl radical is difficult since the stability of a primary carbocation is low.
<img alt="Figure2" src="/files/ftp_upload/13041/13041-PT_Figure_2.svg" style="text-align: center; height: 105px;" />
Figure 2. Fragmentation of 2,2-dimethylpentane.
As shown in Figure 2, the fragmentation of 2,2-dimethyl pentane involves the loss of either methyl or propyl radical, resulting in a tertiary carbocation. Here, the stability of the co-produced radical determines the cleaving bond. So, the cleavage that produces a propyl radical is favored, and the signal from the 2-methyl propyl carbocation becomes the base peak.

This lesson delves into the mass spectrometry of branched alkane fragmentation. Branched alkanes possess secondary or tertiary carbon atoms, which ...

Habituation: Studying Infants Before They Can Talk

Source: Laboratories of Nicholaus Noles, Judith Danovitch, and Cara Cashon&#8212;University of Louisville
Infants are one of the purest sources of information about human thinking and learning, because they&#8217;ve had very few life experiences. Thus, researchers are interested in gathering data from infants, but as participants in experimental research, they are a challenging group to study. Unlike older children and adults, young infants are unable to reliably speak, understand speech, or even move and control their own bodies. Eating, sleeping, and looking around are the only activities babies can perform reliably. Given these limitations, researchers have developed clever techniques for exploring infants&#8217; thoughts. One of the most popular methods makes use of a characteristic of attention called habituation.
Like adults, infants prefer to pay attention to new and interesting things. If they are left in the same environment, over time they become accustomed to their surroundings and pay less attention to them. This process is called habituation. However, the moment something new happens, infants are waiting and ready to pay attention again. This reengagement of attention following habituation is referred to as dishabituation. Scientists can use these characteristic changes in attention as a tool for studying the thinking and learning of young infants. This method involves initially presenting stimuli to infants until they are habituated, and then presenting them with different kinds of stimuli to see if they dishabituate, i.e., notice a change. By carefully choosing the stimuli shown to the infants, researchers can learn a great deal about how infants think and learn.
This experiment demonstrates how habituation can be used to study infant shape discrimination.

Habituação: Estudar bebês antes de poderem falar

Source: Laboratories of Nicholaus Noles, Judith Danovitch, and Cara Cashon&#8212;University of Louisville
Infants are one of the purest sources of ...

Using Your Head: Measuring Infants' Rational Imitation of Actions

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
One of the main challenges of infancy is to learn how to achieve one&#8217;s goals in the world, whether they are to pick up a toy or to express desires to another person, and one of the most powerful tools in this learning process is imitation. However, imitation is not always as simple as copying other people&#8217;s actions; it is also necessary to realize that goals and intentions guide behavior. The world is a complicated place, and the actions that babies imitate are not perfectly presented. For example, consider a baby watching their father drinking from a plastic water bottle. As he picks up the bottle, he accidentally drops it on the floor. He picks it up and dusts it off, before turning the cap and taking a drink. If the baby wants to drink from the bottle on their own, they have to decode this complex set of events and determine which actions are related to their goal. Do they have to drop it and dust it off, or can they simply turn the cap and take a drink?
One way to solve this problem is to view many examples of the same behavior, but some behaviors are rare or different each time they are performed. Thus, it is beneficial for infants to think more about the person they are imitating and less about the specific situation they are observing. If infants assume that others are rational and have goals they are efficiently pursuing, then they can observe a complex event and separate the goal-oriented actions from actions that are incidental or unrelated to their goals. Analyzing the reasons underlying a person&#8217;s behavior allows infants to decide whether it makes sense to imitate that person&#8217;s exact actions or to do things in a simpler or more effective way.
This experiment demonstrates how to measure infant&#8217;s rational imitation of an actor using the methods developed by Meltzoff1 and Gergely, Bekkering, and Kiraly.2

Using Your Head: Measuring Infants&#039; Rational Imitation of Actions

Usando sua cabeça: Medindo a Imitação Racional das Ações dos Bebês

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
One of the main challenges of infancy is to learn how to ...

The Rouge Test: Searching for a Sense of Self

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Humans are different from other animals in many ways, but one of the abilities that sets humans apart is their advanced ability to understand other people and simulate their thoughts and feelings, even when the thoughts and feelings do not align with their own. In scientific terms, these abilities are referred to as theory of mind, and this understanding is necessary for activities like giving compliments, working in groups, asking for favors, and telling white lies.
Humans are not born with a fully developed theory of mind. An individual&#8217;s understanding that they are separate from other people and that they have different desires and knowledge requires an established sense of self. Thus, developing self-recognition and self-awareness are some of the initial steps on the path to developing a mature theory of mind. Studying a child&#8217;s emerging sense of self is complicated, because children&#8217;s conceptual development exceeds their mastery of language. To solve this problem, researchers borrowed methods used to detect self-recognition in animals and applied them to young children. Thus, with a mirror and a bit of make-up, the rouge task was born. 
This video demonstrates how researchers assess self-awareness in children at different ages.

O teste do espelho: em busca de um senso de identidade

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Humans are different from other animals in many ways, but one ...

Numerical Cognition: More or Less

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
One of the goals of the modern education system is to teach children mathematical literacy. They are taught to add, subtract, multiply, and divide, and this base knowledge is used to support learning about geometry, algebra, calculus, physics, and statistics. School-aged children usually acquire these skills in formal educational settings, but the foundation of mathematical understanding is developed much earlier in life.
As infants, humans begin to form the rough representations that allow them to make judgments about number, and perhaps the first numerical concept that humans develop is the idea of less versus more. However, probing these concepts can be challenging, because even if babies have some understanding of number, they have very few ways of showing off what they know. What they can do is crawl, eat, cry, and sleep. Thus, researchers developed a task using this limited set of responses to investigate whether babies can mentally represent number.
This experiment demonstrates how researchers can creatively use food to study concepts of numerical cognition in infants using the method by Feigenson, Carey, and Hauser.1

Investigating Numerical Cognition More/Less in Infants

Cognição Numérica: Mais ou Menos

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
One of the goals of the modern education system is to teach ...

Mutual Exclusivity: How Children Learn the Meanings of Words

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Humans are different from other animals in many ways, but perhaps the most important differentiating factor is their ability to use language. Other animals can communicate and even understand and use language in limited ways, but trying to teach human language to a chimp or a dog takes a great deal of time and effort. In contrast, young humans acquire their native language easily, and they learn linguistic rules without explicit instruction, which is an accomplishment that even the smartest animals cannot match.&#160;
One advantage young humans have over animals is that the human brain is especially adapted to learn new words. With only a few exposures, young children can learn new words and remember them. Perhaps more impressively, children can use what they already know to guide their future learning. For example, children treat objects as if they have only one label. So, if a child has learned the word hammer, they won&#8217;t assume an unfamiliar tool has the same name. This is the principle of mutual exclusivity.1-2
This video demonstrates children&#8217;s ability to use mutual exclusivity to match words to objects in their environment.

Exclusividade Mútua: Como as crianças aprendem os significados das palavras

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Humans are different from other animals in many ways, but ...

How Children Solve Problems Using Causal Reasoning

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Imagine a young child hears an adult say, &#8220;I want to watch the news,&#8221; then watches the adult press a button on the remote control. A moment later, the television screen turns on. The next day, the child wants to turn on the television screen to watch cartoons. How does the child know what to do? Is it enough to say, &#8220;I want to watch cartoons,&#8221; or is pushing the button on the remote control necessary, too? Solving this problem requires children to use the information they observed (i.e., the adult&#8217;s behaviors) to come up with a solution. In their daily lives, children encounter many situations where they need to decode cause-and-effect from complex or ambiguous observations in order to accomplish a goal.
In order to examine children&#8217;s capacity for causal reasoning, psychologists set up tasks using causal scenarios to observe how children draw conclusions and test new hypotheses about the relationships between different types of objects. In these tasks, children are shown interactions involving individual objects or sets of objects. Then, they are asked to identify and use the links between the causes and the effects to solve a problem.
This video demonstrates how to measure children&#8217;s causal reasoning about novel objects based on the methods developed by Gopnik and Sobel1 and Gopnik, Sobel, Schulz, and Glymour.2

Como as crianças resolvem problemas usando raciocínio causal

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Imagine a young child hears an adult say, &#8220;I want to ...

Metacognitive Development: How Children Estimate Their Memory

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Human memory is fallible, and people often cannot accurately recall what they have seen or heard. Adults are aware of their limited memory capacity, so they use strategies, such as rehearsal and mnemonic devices, to improve their recall of important information. Because adults understand the limits of memory, they know it makes more sense to write down the items on their shopping list rather than to try to remember the items when they get to the store. This ability to think about one&#8217;s own memory is called metamemory. Metamemory is one component of a broader set of cognitive processes that allow humans to think about their own knowledge and thinking, which is called metacognition.
Although young children understand that people have thoughts and a limited amount of knowledge, they often have trouble acknowledging the limits of their own knowledge and cognitive skills. Children&#8217;s ability to accurately estimate their own memory capacity and abilities improves over the elementary school years. One common way of measuring metamemory and its development is by giving children an opportunity to predict how well they can remember something, and then observing how well their prediction matches their actual recall.
This experiment demonstrates how to measure children&#8217;s metamemory based on the methods developed by Shin, Bjorklund, and Beck.1

Desenvolvimento Metacognitivo: como as crianças estimam sua memória

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Human memory is fallible, and people often cannot accurately ...

Executive Function and the Dimensional Change Card Sort Task

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Infants are born with amazing cognitive resources at their disposal, but they don&#8217;t know how to use them effectively. In order to harness the power of their brains, humans must develop high-level cognitive processes that manage basic brain functions. These processes make up what psychologists refer to as executive function. Executive function is a key factor in many self-regulatory behaviors, including forming plans to solve problems, negotiating between desires and actions, and directing attention. For example, a child must use several executive processes to stop playing with toys and start cleaning their room. These processes include inhibition (to stop what they&#8217;re doing), planning (to determine what actions need to be performed to clean the room), and attentional control (to stay on task until the cleaning is done). A breakdown of executive function during any of these steps would lead to the room remaining dirty.
Developing executive function is one of the key challenges faced by children as they mature. Some elements of executive function can only be mastered with practice, and brain areas linked to executive function, specifically the prefrontal cortex, develop slowly throughout development, continuing to grow and organize until an individual reaches their twenties. Early demonstrations of executive function have been linked to self-control and behavioral outcomes in children, as well as successes later in life. Relatedly, executive function is impaired in children diagnosed with attention-deficit hyperactivity disorder (ADHD) and autism spectrum disorders.
This experiment demonstrates how to assess executive function in children using the Dimensional Change Card Sort Task, developed by Dr. Philip Zelazo and colleagues.1

Dimensional Change Card Sort Task and Executive Function in Children

Função executiva e a tarefa de classificação do cartão de mudança dimensional

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
Infants are born with amazing cognitive resources at their ...

Categories and Inductive Inferences

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
It might be possible for the human brain to keep track of each individual person, place, or thing encountered, but that would be a very inefficient use of time and cognitive resources. Instead, humans develop categories. Categories are mental representations of real things that can be used for a variety of purposes. For example, individuals can use the perceptual features of animals to place them into a given category. So, upon seeing a furry, four-legged, tail-wagging, barking animal, a person can determine that it is a dog. This is one of many examples where people use perceptual similarity to fit new experiences into their existing mental representations.
However, category membership is much more than skin deep, especially for representations of animals. Frank Keil demonstrated this by using a simple, yet powerful technique that focused on the differences between natural kinds and artifacts. Natural kinds include animals and other living things, while artifacts consist largely of nonliving things, such as tables or gold bricks. In his study, Keil told children stories about natural kinds and artifacts that underwent transformations causing them to cross categorical boundaries. For example, he described a step-by-step process by which a raccoon was transformed into a creature that resembled a skunk in every way. At the end of the story, the raccoon was black with a white stripe, and it had implanted glands that made it smell like a skunk, too. He asked the children to determine whether the resulting animal was a raccoon or a skunk. He used a similar method to describe the transformation of a tire-an artifact-into a shoe. Children&#39;s responses revealed interesting developmental changes into how people think about artifacts and natural kinds.
This video demonstrates Frank Keil&#39;s transformation study.1

How Children Categorize Objects and Make Inductive Inferences

Categorias e Inferências Indutivas

Source: Laboratories of Nicholaus Noles and Judith Danovitch&#8212;University of Louisville
It might be possible for the human brain to keep track of each ...

The Costs and Benefits of Natural Pedagogy

Source: Laboratories of Nicholaus Noles, Judith Danovitch, and Asheley Landrum&#8212;University of Louisville
Children have many tools they use over the course of development to learn from adults. Perhaps the earliest tool is imitation, simply copying what they see an adult do or say. However, children actually learn much more effectively than one might expect if they were only imitating. This is because, when it comes to learning and teaching, children and adults have a special relationship. Children treat adults as if they are helpful and knowledgeable teachers, and adults teach children information in a manner that is usually efficient and effective. Through these interactions, children can learn much better than if they were simply using trial-and-error or copying adults exactly. This way of interacting is referred to as natural pedagogy, and it is one of the reasons that young humans are gifted learners.
One of the most impressive aspects of natural pedagogy is that no one teaches adults how to be good teachers, and children treat adults as teachers without having to be trained to do so. However, natural pedagogy also entails costs. Children are curious and intrinsically motivated to explore, so children do some of their best learning when given opportunities to learn and explore on their own. Thus, the result of natural pedagogy is that children learn information taught to them very effectively, but explicit teaching restricts their curious, exploratory behaviors. There is a tradeoff between efficient learning and self-driven exploration.
This video demonstrates the method by which Elizabeth Bonawitz, Patrick Shafto, and colleagues1 showed the effects of natural pedagogy on young learners.

Investigating the Benefits and Limitations of Natural Pedagogy

Os Custos e Benefícios da Pedagogia Natural

Source: Laboratories of Nicholaus Noles, Judith Danovitch, and Asheley Landrum&#8212;University of Louisville
Children have many tools they use over the ...

Piaget's Conservation Task and the Influence of Task Demands

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Jean Piaget was a pioneer in the field of developmental psychology, and his theory of cognitive development is one of the most well-known psychological theories. At the heart of Piaget&#8217;s theory is the idea that children&#8217;s ways of thinking change over the course of childhood. Piaget provided evidence for these changes by comparing how children of different ages responded to questions and problems that he designed.
Piaget believed that at age 5, children lack mental operators or logical rules, which underlie the ability to reason about relationships between sets of properties. This characteristic defined what he called the preoperational stage of cognitive development. One of Piaget&#8217;s classic measures of children&#8217;s ability to use mental operations is his conservation task. In this task, children are shown two identical objects or sets of objects. Children are first shown that the objects are the same on one key property (number, size, volume, etc.). Then, one of the objects is modified so it appears different than the other one (e.g., it is now longer, wider, or taller), but the key property remains the same. Following this transformation, children are asked to judge if the two objects or sets of objects are now the same or different with respect to the original key property.
Piaget reported that children in the preoperational stage (approximately ages 2-7) typically judged the objects to be different after the transformation, even though the key property had not changed. He attributed children&#8217;s incorrect responses to their excessive focus on the change, rather than on the fact that the key property remained the same. However, over the years, researchers have argued that Piaget&#8217;s conservation task is an invalid measure of children&#8217;s reasoning skills. These critics have suggested that children&#8217;s poor performance is due to task demands, such as assumptions about the experimenter&#8217;s goals and expectations when the question about the key property is repeated.
This video demonstrates how to conduct Piaget&#8217;s classic conservation task,1&#8211;2 and how a small modification in the task design can dramatically change children&#8217;s accuracy (based on the methods developed by McGarrigle and Donaldson3).

Piaget&#039;s Conservation Task and the Influence of Task Demands

A Tarefa de Conservação de Piaget e a influência das demandas de tarefas

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Jean Piaget was a pioneer in the field of developmental ...

Children's Reliance on Artist Intentions When Identifying Pictures

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Children are not the best artists. Sometimes it&#8217;s easy to pick out the characteristic triangular head, whiskers, and tail of a cat, but children often describe elaborate scenarios that they depict as a beautifully unrecognizable mess. Thus, given children&#8217;s questionable artistic talent, how do they know what their drawings, and the drawings of others, represent? One way children identify pictures is by relying on resemblance. If it looks like a cat, then it&#8217;s a cat. However, some pictures do not clearly resemble any real object. In this situation, children must use other means to figure out what the picture represents, including their understanding of what the person who created the picture intended it to represent.
By their first birthday, children are sensitive to the intentions of other people. They know that people&#8217;s actions are driven by their goals, and they can infer a person&#8217;s intentions even if the goal-directed action is not successful (e.g., they understand a person struggling to turn a lid intends to open a jar, even if they never see them succeed in opening it). By about age 3, children can use this understanding of intention to guide their interpretation of drawings and other pictorial representations. They apply this understanding both to identifying their own drawings, and to interpreting drawings created by another person.
This experiment demonstrates how to measure children&#8217;s use of intention to interpret otherwise ambiguous pictures based on the methods developed by Bloom and Markson.1

Children&#039;s Reliance on Artist Intentions When Identifying Pictures

Confiança das crianças nas intenções dos artistas ao identificar imagens

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Children are not the best artists. Sometimes it&#8217;s easy ...

Measuring Children's Trust in Testimony

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
How does a person learn about the world around them? One way is through direct observation and exploration. However, not every piece of information can be observed firsthand. Instead, a person must often rely on other people as information sources. This is particularly true for children who have so many questions about the world around them, yet have limited means of accessing the answers. Thus, children must rely on other people to provide answers to their questions.
There is a popular viewpoint that children are gullible and that they believe everything they hear. However, recent research has shown this is not the case. Children as young as age 3 evaluate what other people say and show selective trust in other people&#8217;s testimony. Children pay attention to and use their knowledge about an individual&#8217;s prior behavior and characteristics to judge whether that individual is a trustworthy informational source. &#160;
This video demonstrates how to measure children&#8217;s trust in testimony based on methods developed by Birch, Vauthier, and Bloom1 and Koenig, Clement, and Harris.2&#160;

Measuring Children&#039;s Trust in Testimony

Medindo a confiança das crianças em depoimento

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
How does a person learn about the world around them? One way ...

Are You Smart or Hardworking? How Praise Influences Children's Motivation

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Imagine teaching two children how to skate. It is a hard task for both of them, and they fall down frequently. After falling down for the first time, one child says that skating is too hard and wants to go home. The other child seems to enjoy the challenge and eagerly gets back up after falling down each time. Why do the children have such different attitudes about the same task? One reason may be that they have different mindsets or beliefs about the nature of their ability.
According to psychologist Carol Dweck, some people have a fixed mindset, and some people have a growth mindset. People with a fixed mindset believe that intelligence or abilities are fixed and cannot change. When these people face a challenge, like learning how to skate, they tend to believe that if a new skill does not come easily, then they are simply no good at it. They do not see their skills as capable of changing, and thus they decide that it&#8217;s useless to continue trying. People with a growth mindset have the opposite attitude. They believe that abilities can be developed through hard work, and they continue trying to improve even if they do not initially succeed.
How do these different mindsets develop? One factor that influences children&#8217;s persistence and motivation to succeed is the way their success is described by other people. Specifically, the kind of praise children receive from adults, such as parents and teachers, can have a powerful effect on their subsequent motivation to perform a challenging task.
This video demonstrates how to measure the effect of praise on children&#8217;s motivation based on the methods developed by Mueller and Dweck.1

Investigating How Praises Influence Children&#039;s Motivation

Você é inteligente ou trabalhador? Como o elogio influencia a motivação das crianças

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
Imagine teaching two children how to skate. It is a hard task ...

Memory Development: Demonstrating How Repeated Questioning Leads to False Memories

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
A person is defined as a unique individual based on the people and events they encounter in their lives. Thus, creating, storing, and recalling memories are essential elements of the human experience. However, memory, as adults experience it, takes time to develop. Although young children can learn facts and remember details of their lives from moment-to-moment and day-to-day, they do not create autobiographical memories or detailed memories of events that happen in their lives until age 3 or older.
Even after age 3, children&#8217;s memories differ from those of adults in important ways. Children are less effective at evaluating their own memories than adults, which makes it difficult for them to determine, for example, whether or not their memories are accurate. False memories are a problem for both children and adults, as it is quite easy to create a false memory with a poorly-worded question or a story repeated over and over. However, young children are more susceptible to creating false memories than either older children or adults.
This video demonstrates children&#8217;s vulnerability to false memories using a method developed by Steven Ceci and his collaborators.1-2

Demonstrating How Repeated Questioning Leads to False Memories

Desenvolvimento da memória: demonstrando como o questionamento repetido leva a falsas memórias

Source: Laboratories of Judith Danovitch and Nicholaus Noles&#8212;University of Louisville
A person is defined as a unique individual based on the people ...

Education Developmental Psychology

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Education Developmental Psychology

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