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Research

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 ...

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: 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 ...

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 ...

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 ...

&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 ...

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 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 ...

&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 ...

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 ...

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 ...

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 ...

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 ...

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: 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: 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: 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: 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: 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: 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: Long-Chain Alkane Fragmentation

The molecular ions of linear alkanes prefer to fragment at the carbon-carbon bond away from the end of the chain since the cleavage of an inner bond creates a stable carbocation and a stable radical. Consequently, the mass signals of linear alkanes feature intense peaks in the middle of the mass-to-charge ratio plot with weaker peaks on either end. The fragmentation of each carbon-carbon bond with the release of a methyl group in each splitting leads to prominent peaks in the mass spectra separated by 14 u. Smaller mass signals near each prominent peak result arise from secondary fragmentation at the carbon-hydrogen bond.
For example, consider the fragmentation pattern of n-hexane shown in Figure 1 and the corresponding mass spectra in Figure 2.
<img alt="Figure1" src="/files/ftp_upload/12299/12299-PT_Figure_1.svg" style="text-align: center; height: 500px;" />
Figure 1: Fragmentation of n-hexane molecular ion.
<img alt="Figure2" src="/files/ftp_upload/12299/12299-PT_Figure_2.svg" style="text-align: center; height: 275px;" />
Figure 2: Mass spectrum of n-hexane.
The signals at a mass-to-charge ratio of 71, 57, 43, 29, and 15 indicate that the cleavage can occur at all carbon-carbon bonds in the molecule. However, the relative abundance of these peaks varies. The difference between molecular ion signal and signal at 71 indicates the separation of methyl groups (M.W. = 15 u) group from molecular ions. Further release of a CH2 group from that fragment (M.W. = 14 u) results in a signal at 57. The signals at 43, 29, and 15 are due to the subsequent releases of CH2 groups. The abundance of signals at 15 and 71 is minimal due to the least probability for the formation of unstable methyl carbocation or a methyl radical, respectively. The base peak at 43 indicates the most preferred fragmentation, which yields a pair of stable carbocation and radical.

The molecular ions of linear alkanes prefer to fragment at the carbon-carbon bond away from the end of the chain since the cleavage of an inner bond ...

Material Constants

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In contrast to the production of cars or toasters, where millions of identical copies are made and extensive prototype testing is possible, each civil engineering structure is unique and very expensive to reproduce (Fig.1). Therefore, civil engineers must extensively rely on analytical modeling to design their structures. These models are simplified abstractions of reality and are used to check that the performance criteria, particularly those related to strength and stiffness, are not violated. In order to accomplish this task, engineers require two components: (a) a set of theories that account for how structures respond to loads, i.e., how forces and deformations are related, and (b) a series of constants that differentiate within those theories how materials (e.g. steel and concrete) differ in their response.
<img alt="Figure 1" src="/files/ftp_upload/10363/10363fig1v2.jpg" /> 
Figure 1: World Trade Center (NYC) transportation hub.
Most engineering design today uses linear elastic principles to calculate forces and deformations in structures. In the theory of elasticity, several material constants are needed to describe the relationship between stress and strain. Stress is defined as the force per unit area while strain is defined as the change in dimension when subjected to a force divided by the original magnitude of that dimension. The two most common of these constants are the modulus of elasticity (E), which relates the stress to the strain, and Poisson&#39;s ratio (&#957;), which is the ratio of lateral to longitudinal strain. This experiment will introduce the typical equipment used in a construction materials laboratory to measure force (or stress) and deformation (or strain), and use them to measure E and &#957;&#160;of a typical aluminum bar.

Elasticity Theory; Modulus of Elasticity and Poisson&#039;s Ratio

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In contrast to the production of cars or ...

Stress-Strain Characteristics of Steels

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
The importance of materials to human development is clearly captured by the early classifications of world history into periods such as the Stone Age, Iron Age, and the Bronze Age. The introduction of the Siemens and Bessemer processes to produce steels in the mid-1800s is arguably the single most important development in launching the Industrial Revolution that transformed much of Europe and the USA in the second half of the 19th century from agrarian societies into the urban and mechanized societies of today. Steel, in its almost infinite variations, is all around us, from our kitchen appliances to cars, to lifelines such as electrical transmission networks and water distribution systems. In this experiment we will look at the stress-strain behavior of two types of steel that bound the range usually seen in civil engineering applications - from a very mild, hot rolled steel to a hard, cold rolled one.

Uniaxial Tensile Test and Stress-Strain Characteristics of Steels

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
The importance of materials to human ...

Stress-Strain Characteristics of Aluminum

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Aluminum is one of the most abundant materials in our lives, as it is omnipresent in everything from soda cans to airplane components. Its widespread use is relatively recent (1900AD), primarily because aluminum does not occur in its free state, but rather in combination with oxygen and other elements, often in the form of Al2O3. Aluminum was originally obtained from bauxite mineral deposits in tropical countries, and its refinement requires very high-energy consumption. The high cost of producing quality aluminum is another reason why it is a very widely recycled material.
Aluminum, especially when alloyed with one or more of several common elements, has been increasingly used in architectural, transportation, chemical, and electrical applications. Today, aluminum is surpassed only by steel in its use as a structural material. Aluminum is available, like most other metals, as flat-rolled products, extrusions, forgings, and castings. Aluminum offers superior strength-to-weight ratio, corrosion resistance, ease of fabrication, non-magnetic properties, high thermal and electrical conductivity, as well as ease of alloying.

Uniaxial Tensile Strength and Stress-Strain Test of Aluminum

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Aluminum is one of the most abundant materials ...

Charpy Impact Test of Cold Formed and Hot Rolled Steels Under Diverse Temperature Conditions

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
One of the more insidious types of failures that can occur in structures are brittle fractures,which are mostly due to either poor quality materials or poor material selection. Brittle fractures tend to occur suddenly and without much material inelasticity; think of a bone fracture, for example. These failures often occur in situations where there is little ability for the material to develop shear stresses due to three-dimensional loading conditions, where local strain concentrations are high, and where a logical and direct force path was not provided by the designer. Examples of this type of failure were observed in the aftermath of the 1994 Northridge earthquake in multi-story steel structures. In these buildings, a number of the key welds fractured without displaying any ductile behavior. Fractures tend to occur near connections, or at interfaces between pieces of base materials, as welding tends to introduce local discontinuities in both, materials and geometry, as well as three-dimensional stresses due to cooling.
When specifying materials for a structure that will see very low operating temperatures (i.e., the Alaska pipeline) many cycles of loading (a bridge on an interstate highway), or where welding is used extensively, it is necessary to have a simple test that characterizes the material&#39;s robustness, or resistance to fracture. In the civil engineering field that test is the Charpy V-notch test, which is described in this lab. The Charpy V-notch test is intended to provide a very simplistic measure of the material&#39;s ability to absorb energy when subjected to an impact load.

Charpy Impact Test of Cold Formed and Hot Rolled Steels

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
One of the more insidious types of failures ...

Rockwell Hardness Test and the Effect of Treatment on Steel

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Hardness testing is one of the most universally valuable mechanical tests available to engineers, as it is both simple and relatively inexpensive for the wealth of information and data it produces. Hardness testing, generally in the form of a surface penetration test, is both quicker and less destructive than tensile testing. Hardness provides a linear relationship with tensile strength over a wide range of strengths for many materials, such as steel. Hardness tests are empirical, rather than derived from theory, as the results conflate effects from many different materials properties (Young&#39;s modulus, yield strength, etc.).
Hardness is a characteristic of a material used to describe how much plastic deformation (yield) that a material will undergo when a known force is applied). One can characterize hardness in three manners: scratch, indentation, and rebound hardness. A common early example of a hardness (scratch) test is the Mohs scale (1820), derived for minerals, and in which talc has a value of 1 and diamond a value of 10. In indentation testing using the Rockwell approach, small indenters are used with different loads. The most common are the Rockwell Hardness B (HRB), which uses a 1/16 in. hardened steel ball indenter along with a 100 kg weight, and the Rockwell Hardness C (HRC), which uses a diamond cone indenter along with a 150 kg weight. HRB tests are conducted for materials with low range hardness, such as aluminum, brass, and soft steels, whereas HRC tests are used for materials with high range hardness, such as harder steels. Smaller weights (15 to 45 kg) are used for the Rockwell superficial hardness test such as the HR15W, which uses a 1/8 in. steel ball with a 15 kg weight. With its lower load and shallow impression, a superficial Rockwell hardness test is ideal for very thin or brittle materials. An example of a rebound test is the Schmidt hammer, used to measure the strength of concrete. In this test, a steel mass is shot at the surface with a known force and the rebound of the ball is measured. In all types of hardness testing, it is imperative to conduct extensive calibrations if reliable results are to be obtained.
For this experiment, we will be examining the Rockwell hardness test, which measures indentation hardness of untreated and heat-treated aluminum.

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Hardness testing is one of the most ...

Buckling of Steel Columns

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In the design of civil works, it is important to deliver structures that are not only safe under unexpected loads, but also provide excellent performance under everyday loads at a reasonable economic cost. The latter is often tied to minimum use of materials, ease of fabrication, and rapid construction in the field. Structures made of steel members can be very economical because of the great strength of the material and the extensive prefabrication of their members and connections, which help maximize the speed of construction on site. Generally, the skeleton of a steel structure will be very slender as compared to a reinforced concrete one. While its behavior in tension is governed primarily by the strength of the material, steel in compression is governed by another failure mode common to all materials- buckling. This behavior is easily demonstrated by pressing down on a slender wooden ruler, which under a compressive load will suddenly move sideways and lose load carrying capacity. This phenomenon will occur in any slender member of a structure. In this lab, we will measure the buckling capacity of a series of slender aluminum columns to illustrate this failure mode, which over time has led to many catastrophic failures including that of the Quebec River Bridge, which was erected in 1918.

Steel Column Buckling and Measuring the Buckling Capacity

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In the design of civil works, it is important ...

Dynamics of Structures

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
It is rare nowadays that a whole year goes by without a major earthquake event wreaking havoc somewhere around the world. In some cases, like the 2005 Banda Ache earthquake in Indonesia, the damage involved large geographic areas and casualties in the six figures. In general, the number and intensity of earthquakes is not increasing, however, the vulnerability of the built environment is rising. With increasing unregulated urbanization around seismically active areas, such as the Circum-Pacific &#34;belt of fire,&#34; sea rising in low-laying coastal area, and increasing concentrations of both energy production/distribution and digital/telecommunication network critical nodes in vulnerable areas, it is clear that earthquake-resistant design is key to future community resilience.
Designing structures to resist earthquake damage has progressed greatly in the last 50 years, primarily through work in Japan following the 1964 Niigata Earthquake, and in the United States following the 1971 San Fernando Valley Earthquake. The work has advanced along three parallel tracks: (a) experimental work aimed at developing improved construction techniques to minimize damage and loss of life; (b) analytical studies based on advanced geometrical and non-linear material models; and, (c) synthesis of the results in (a) and (b) into design code provisions that improve the ability of structures to resist unexpected loads.
Seismic testing in a laboratory setting is often difficult and expensive. Testing is primarily carried out using the following three techniques:
<ol>
	<li class="jove_content">Quasi-static testing (QST), where parts of a structure are tested using slowly applied and equivalently predetermined lateral deformations with idealized boundary conditions. This technique is particularly useful to assess the effects of structural detailing on the toughness and deformation capacity of particular parts of structures.</li>
	<li class="jove_content">Pseudo-dynamic testing (PSDT), where loads are also applied slowly, but the dynamic effects are taken into account by solving the equations of motion as the test progresses and by utilizing direct test feedbacks (primarily the instantaneous stiffness) to assess the actual stiffness and damping characteristics of the structure.</li>
	<li class="jove_content">Shake tables, where scale models of complete structures are subjected to input motions using a hydraulically actuated base or foundation. Shake tables represent a more faithful testing technique, as the structure is not artificially restrained, the input is true ground motion, and the resulting forces are truly inertial ones, as one would expect in a real earthquake. However, the power requirements are enormous, and only a few shake tables capable of working at nearly full-scale exist around the world. Globally, there is only one large shake table capable of carrying out tests on full-scale structures, which is the shake table at the E-Defense facility in Japan, built in the aftermath of the 1985 Kobe earthquake.</li>
</ol>
In this experiment, we will utilize a small shake table and model structures to study the dynamic behavior characteristics of some structural models. It is these dynamic characteristics, principally the natural frequency and damping, as well as the quality of the structural detailing and construction, which make structures more or less vulnerable to earthquakes.

Quasi-static Testing, Pseudo-dynamic Testing and Shake Tables

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
It is rare nowadays that a whole year goes by ...

Fatigue of Metals

Source: Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
The importance of studying metal fatigue in civil infrastructure projects was brought into the spotlight by the collapse of the Silver Bridge in Point Pleasant, West Virginia in 1967. The eyebar-chain suspension bridge over the Ohio River collapsed during evening rush hour, killing 46 people as a result of the failure of a single eyebar with a small 0.1-inch defect. The defect reached a critical length after repeated loading conditions and failed in a brittle fashion causing the collapse. This event garnered attention in the bridge engineering community and highlighted the importance of testing and monitoring fatigue in metals.
Under normal service conditions, a material can be subjected to numerous applications of service (or everyday) loads. These loads are typically at most 30%-40% of the ultimate strength of the structure. However, after the accrual of repeated loadings, at magnitudes substantially below the ultimate strength, a material can experience what is termed fatigue failure. Fatigue failure can occur suddenly and without significant prior deformation and is linked with crack growth and rapid propagation. Fatigue is a complex process, with many factors affecting fatigue resistance (Table 1). This complexity underscores the integral need for routine and thorough inspection of structures subjected to repeated loadings such as bridges, cranes, and almost all types of vehicles and aircrafts.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Stressing conditions</td>
			<td>Material properties</td>
			<td>Environmental conditions</td>
		</tr>
		<tr>
			<td>
			<ul>
				<li>Type of stress</li>
				<li>Stress amplitude</li>
				<li>Mean stress</li>
				<li>Frequency</li>
				<li>Combined stresses</li>
				<li>Stress history</li>
				<li>Stress concentrations (notches)</li>
				<li>Rolling contact</li>
				<li>Size</li>
			</ul>
			</td>
			<td>
			<ul>
				<li>Type of material</li>
				<li>Surface conditions</li>
				<li>Grain size</li>
			</ul>
			</td>
			<td>
			<ul>
				<li>Temperature&#160;</li>
				<li>Corrosion</li>
			</ul>
			</td>
		</tr>
	</tbody>
</table>
Table 1. Factors affecting fatigue

Metal Fatigue and Testing Cycles to Failure

Source: Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
The importance of studying metal fatigue in civil ...

Tension Tests of Polymers

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Polymeric materials are widely used in civil structures, with uses ranging from very soft sealants to more rigid pipes in water and wastewater systems. The most basic definition of a polymer is a molecular structure with repeating subunits. The term polymer comes from Greek, where &#34;poly&#34; means many, and &#34;-mer&#34; means basic unit. Monomers, or single mers, are the specific repeating units. With polymers, the structure, including the length of the carbon backbone and the varying flexibility, will dictate the properties of the polymer. Polymers are classified into 3 subcategories: plastics, elastomers, and rigid rod polymers. Plastics are further subdivided into thermosets, which do not soften on heating, and thermoplastics, which do soften when heated and harden on cooling. Additionally, thermoplastics are mostly linear or branched polymers with little to no cross-linking, whereas thermosets exhibit 3D structure and have extensive cross-linking. Elastomers, or rubbers, are long, coiled chains and can be stretched to twice the original length, but will contract back to the original size when released, whereas rigid rod polymers do not stretch and are strong, crystalline structures.
In this laboratory, we will look at several different polymeric materials, including high density polyethylene (HDPE), polyvinyl chlorides (PVC), nylon, and methyl methacrylate (acrylic) in order to understand the breadth and diversity of the stress-strain curves for these materials and how their mechanical properties affect their performance.

Uniaxial Tensile Test Strength for Polymers

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Polymeric materials are widely used in civil ...

Tension Test of Fiber-Reinforced Polymeric Materials

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Fiber-reinforced polymeric materials (FRP) are composite materials that are formed by longitudinal fibers embedded in a polymeric resin, thereby creating a polymer matrix with aligned fibers along one or more directions. In its simplest form, the fibers in FRP materials are aligned in an orderly, parallel fashion, thus imparting orthotropic material characteristics, meaning that the material will behave differently in the two directions. Parallel to the fibers, the material will be very strong and/or stiff, whereas perpendicular to the fibers will be very weak, as the strength can only be attributed to the resin instead of the whole matrix.
An example of this unidirectional configuration is the commercially available FRP reinforcing bars, which mimic the conventional steel bars used in reinforced concrete construction. FRP materials are used both as stand-alone structures such as pedestrian bridges and staircases, and also as materials to strengthen and repair existing structures. The thin, long plates are often epoxied to existing concrete structures to add strength. In this case, the FRP bars act as external reinforcement. The FRP bars and plates are lighter and more corrosion resistant, so they are finding applications in bridge decks and parking garages, where de-icing slats lead to rapid deterioration of conventional bars.
In this laboratory exercise, the tensile behavior of a unidirectional specimen will be studied, with emphasis on its ultimate strength and deformation capacity. The behavior of the specimen is expected to be elastic until failure, which is expected to occur in a sudden and explosive manner. This behavior should be contrasted with those of ductile steels, which exhibit extensive deformation capacity and strain hardening before failure.

Uniaxial Tension Test of Fiber-Reinforced Polymeric Materials

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Fiber-reinforced polymeric materials (FRP) are ...

Aggregates for Concrete and Asphaltic Mixes

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Concrete and asphalt are by far the most common construction materials used today. Concrete is a composite material consisting of cement, water, air, coarse aggregate, and fine aggregates. Fine aggregates are typically sands and coarse aggregates are natural or crushed rocks. Chemical admixtures to modify certain specific properties are also commonly used (i.e., superplasticizers to make the concrete fluid during casting). Asphaltic mixes consist primarily of asphalts, coarse aggregates, and fine aggregates, in addition to a number of emulsifiers and other additives used to improve viscosity during placement.
In both concrete and asphaltic mixes, aggregates make up a very significant portion of the mix volume, as economy requires that the amount of cement and asphalt be minimized. Two types of aggregates are commonly recognized: coarse aggregates, defined as particles larger than about 4.75mm (rocks), and fine aggregates, consisting of smaller particles (sands). Other important characteristics of aggregates are that they be rigid, durable, and chemically inert with respect to the concrete mortar or asphalt. Aggregates are intended to be fillers, but they are not intended to play a key role in the behavior of either material. However, the stiffness and strength of the aggregates needs to be higher than the concrete mortar or asphalt, so as not to be the controlling phase.
For effective performance, several characteristics of the aggregates, ranging from their mechanical and chemical properties to their size distribution, need to be taken into consideration in the aggregate mix design. Moreover, both concrete mixes undergo very different behavior when being placed, with the materials resembling a Newtonian fluid, and when in their hardened configuration, with the materials resembling an elastic solid. Additionally in the case of asphalt, the service temperature range is very important, as the properties of asphalts are temperature-dependent within the usual serviceability temperature range.
In this laboratory, we will examine the basic properties of aggregates that are needed to develop successful concrete mix designs. The properties needed for asphalts are very similar, but sometimes utilize different testing techniques. The primary characteristics that we will look at are the size distribution, specific gravity, absorption, moisture content, and bulk density, all of which will be described and measured in this laboratory exercise. Other important characteristics that will not be addressed in this module are the shape and angularity of the particles, abrasion and impact resistance, chemical stability, as well as the soundness and presence of harmful organics.

Sieve Analysis of Aggregates for Concrete and Asphaltic Mixes

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Concrete and asphalt are by far the most ...

Compression Tests on Hardened Concrete

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
There are two distinct stages in a construction project involving concrete. The first stage involves batching, transporting, and casting fresh concrete. At this stage, the material is viscous, and the workability and finishability are the key performance criteria. The second stage occurs when the hydration process begins shortly after the concrete is placed in the form, and the concrete will set and begin to harden. This process is very complex, and not all of its phases are well understood and characterized. Nevertheless, the concrete should achieve its intended design strength and stiffness at about 14 to 28 days after casting. At this point, a series of tests will be conducted on concrete cylinders cast at the time of placement to determine the concrete&#39;s compressive and tensile strengths, as well as on occasion, its stiffness.
The objectives of this experiment are threefold: (1) to conduct compressive cylinder tests to determine the 7-, 14-, and 28-day strength of concrete, (2) to determine the modulus of elasticity at 28 days, and (3) to demonstrate the use of a simple non-destructive test to determine in situ concrete strength.

Compression and Schmidt Hammer Tests on Hardened Concrete

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
There are two distinct stages in a ...

Tests of Hardened Concrete in Tension

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In a previous laboratory focused on concrete in compression, we observed that concrete can withstand very large stresses under uniaxial compressive forces. However, the failures observed were not compressive failures but failures along shear planes where maximum tensile forces occur. Thus, it is important to understand the behavior of concrete in tension and particularly its maximum strength as that will govern both its ultimate and service behavior. From the ultimate standpoint, combinations of tension and shear stresses will lead to cracking and immediate and catastrophic failure. For that reason, concrete is seldom if ever used in an unreinforced condition in structural applications; most concrete members will be reinforced with steel so that these cracks can be stopped and the crack widths limited. The latter is important from the serviceability standpoint because controlling crack widths and distribution is the key to durability, as this will impede deicing salts and similar chemicals from penetrating and corroding the reinforcing steel.
The objectives of this experiment are threefold: (1) to conduct tensile split cylinder tests to determine concrete tensile strength, (2) to conduct beam tests to determine concrete tensile strength, and (3) to demonstrate the influence of steel reinforcement on behavior by comparing the behavior of lightly reinforced beam with an unreinforced one.

Split Tension Test and Beam Tension Test for Hardened Concrete

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
In a previous laboratory focused on concrete ...

Tests on Wood

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Wood is a ubiquitous material that has been used in construction from the earliest times. Wood is a renewable, sustainable material with great aesthetic value. Today, there are probably more buildings constructed with wood than any other structural material. Many of these buildings are singlefamily residences, but many larger apartment buildings, as well as commercial and industrial buildings, also use wood framing.
The widespread use of wood in construction has appeal from both an economic and aesthetic basis. The ability to construct wood buildings with a minimal amount of equipment has kept the cost of woodframe buildings competitive with other types of construction. On the other hand, where architectural considerations are important, the beauty and warmth of exposed wood is difficult to match with other materials.
The objectives of this experiment are to conduct tensile and compressive tests on three types of wood to investigate their stress-strain behavior, and to conduct a four-point bending test on a wood beam to ascertain its flexural performance. In a four-point bending test, a simply-supported beam is loaded with two equal point loads at its third points, resulting in a central portion with constant moment and zero shear. This is an important test because wood structural elements are often used in floor systems and are thus primarily loaded by bending stresses.

Compression Tests, Tension Tests and Bending Tests on Wood

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Wood is a ubiquitous material that has been ...

Tests on Fresh Concrete

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Concrete consists of two phases, a cement paste phase comprised of cement, water, and air, and an aggregate phase comprised of coarse and fine aggregates. There are two key considerations when designing a concrete mix. First, the concrete must be workable and easy to cast in the forms in its fresh condition, even when the forms are packed with steel reinforcement. In this condition, it is the rheology of concrete that is important. Second, the mix must produce a hardened concrete of specified strength at 28 days (or similar specified time) that is durable and provides good serviceability.
In this laboratory exercise, a method of concrete mixture proportioning, named the trial batch method, will be explored. The concrete produced will be used in conducting typical tests to determine the principal characteristics of fresh concrete, including slump, flowability, air content, and density. The trial batch method is a simple, empirical approach to mixture design.
The objectives of this experiment are fourfold: (1) to use the trial batch mix method to determine optimum proportions of aggregates, cement, and water for concrete to meet specified slump requirements, (2) to learn concrete mixing practice in a laboratory environment, (3) to observe the characteristic properties of fresh concrete, and (4) to prepare 4&#34;x8&#34; concrete cylinders for subsequent evaluation.

Testing Fresh Concrete by the Trial Batch Method

Source:&#160;Roberto Leon, Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA
Concrete consists of two phases, a cement ...

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