0 Bewertungen0% fanden dieses Dokument nützlich (0 Abstimmungen)

53 Ansichten13 Seitennmr relaxation uiuc

Sep 21, 2017

© © All Rights Reserved

PDF, TXT oder online auf Scribd lesen

nmr relaxation uiuc

© All Rights Reserved

Als PDF, TXT **herunterladen** oder online auf Scribd lesen

0 Bewertungen0% fanden dieses Dokument nützlich (0 Abstimmungen)

53 Ansichten13 Seitennmr relaxation uiuc

© All Rights Reserved

Als PDF, TXT **herunterladen** oder online auf Scribd lesen

Sie sind auf Seite 1von 13

Experiment 6:

Determination of NMR relaxation times

Overview

The goal of this lab is to measure the longitudinal and transverse relaxation times of a

Quinine/CDCl3 sample. Quinine is a molecule that has a variety of uses in medicine. The sample

is a standard that is kept in the 145 RAL, which is the location of the UIUC NMR lab. There

will be no sample preparation required and the TA will perform the procedure, however you are

expected to come prepared to observe and ask questions. This experiment is designed to provide

insight into the use of NMR spectroscopy to study important chemical properties.

You MUST sign up for a time slot with the TA during the NMR mini lecture. You will perform

the experiment on that day and will only report to your regular lab day to turn in assignments.

We will meet in Noyes Lab 157, and then walk together to the NMR facility.

Required Readings

1. Introduction to Nuclear Magnetic Resonance Spectroscopy. University of Illinois. 2014.

http://www.scs.illinois.edu/nmr/handouts/getting_started/NMR-intro-DLO.pdf

2. Inside an NMR Magnet. Massachusetts Institute of Technology. 2014

http://web.mit.edu/speclab/www/Facility/shim-probe-sample.html

3. What is a Probe? University of Colorado. 2014.

http://chemnmr.colorado.edu/moreinfo/instruments/probe_whatis.html

4. Reich H.J., Relaxation in NMR Spectroscopy. University of Wisconsin. 2014.

http://www.chem.wisc.edu/areas/reich/nmr/08-tech-01-relax.htm

1

5. Download MNova according to emailed instructions. Any questions or problems can be

addressed at the mini lecture.

Background

You have likely seen NMR spectroscopy before as a tool for organic structure determination.

While it is true that it is very useful in this setting, NMR spectroscopy can be used for many

other applications as well. These include kinetic rate determination, protein structures and

interactions, temperature studies, and molecular property characterization. In particular, this lab

will focus on molecular property characterization. However, first we must understand the basics

of NMR spectroscopy, and then we can conduct and understand specialized experiments.

Nuclei with non-zero spins align when they are placed in an external magnetic field. When they

align with the field they are said to be in a low energy state, and when they align against the field

they are said to be in a high energy state. A diagram depicting this is shown below in Figure 1.

Figure 1. Energy levels for different spin states are shown. Splitting occurs for nuclei with non-

zero spins. A higher energy indicates an alignment against the magnetic field.

molecules in a strong magnetic field. When a nucleus is exposed to radiation of a suitable

energy, such as from a radiofrequency pulse, it undergoes a transition from the lower-energy-

state to the higher-energy-state. After a relaxation time, these nuclei release the absorbed energy

and return to their lower-energy-state. This relaxation time can be either longitudinal (1 ),

transverse (2 ), or a combination of the two. They are dependent on various system factors and

will be described in detail below. These relaxation times are characteristic of nuclei in different

chemical environments. They can be used to study molecular interactions and dynamics, as well

as be used for characterization. In this lab, we seek new ways of exploring chemical systems

using NMR. For this purpose, we will determine the longitudinal and transverse relaxation times

for a standard sample.

Before relaxation times can be determined, however, the 90o pulse width must be found. It is

normally written as pw90, and represents the length of a radiofrequency pulse, in microseconds,

that produces the maximum emission of energy from a nucleus of interest. An array of pulse

2

width values is created to determine the pw360, or the null pulse. At this point, there is zero

emission from the nucleus of interest. A figure illustrating such an experiment is shown below.

The pw90 is simply found by dividing the pw360 by 4.

Figure 2. Pulse width array. The pw360 is determined because it is the easiest to detect, and the

pw90 is calculated from this value.

1 relaxation occurs when a nucleus in the higher energy state passes precessional energy to the

surrounding material which is referred to as the lattice. The 1 relaxation process is a non-

radiative process, and is possible due to the molecular motion and energy of the lattice which can

interact with the spinning nucleus of the sample. The average amount of time that a nucleus

spends in the higher-energy-state is what we measure as the longitudinal relaxation time. It is

dependent on the identity of the lattice, as well as the magnetogyric ratio of the absorbing

nucleus. Because these processes are all based on molecular motion, 1 is shorter for lattices of

low viscosity and small nuclei with flexible bonds.

1 relaxation can be measured through an inversion-recovery experiment, depicted below in

Figure 3. Essentially, a pulse is applied, which puts the nucleus into the opposite spin state,

and after a delay time, a 2 pulse is applied. The system will naturally undergo relaxation to the

original orientation. However, the intensity of the observed NMR signal is dependent on the

fraction of the molecules still in the excited state.

3

Figure 3. Inversion-recovery experiment2. From this figure, it can be seen that various time

constants of relaxation delay produce different intensity values.

An array of delay times is constructed to determine the null time,1 , which is derived in the

following way. In general,

0

= ( ) (Equation 1)

1

represents the magnetic intensity at a certain time. 0 is the initial magnetic field signal.

Equation 1 can be integrated and solved to gain the magnetization expression dependent on time.

= 0 1 (Equation 2)

In an inversion recovery experiment, the effects of 2 are removed because no energy is lost in

the x-y plane, and the following are true:

(0) = (Equation 3)

= (1 2 1 ) = (1 2 ) (Equation 4)

4

The system can be modeled as a type of exponential fit using the quality on the right side of the

equation. The exponential constant is related to longitudinal relaxation in the following way:

1

= (Equation 5)

1

Once the data are fit using this equation, 1 can be easily determined. However, Microsoft Excel

does not have a natural function to model this system, so we will use the Solver Add-in. To add

this to Microsoft Excel, go to File, Options, Add-ins, Go (near the bottom where it says Manage

Add-ins), and select Solver Add-in. Press okay, and it will be added to the Data tab in the

Analysis section. Solver requires that we set up a system as shown in Figure 4. Enter in the

appropriate data in the first two columns and enter the formulas with the correct cell references

into the remaining columns. We will minimize the Sum of Square Difference by modifying the B

and F cells. Plot the Formula values as a trendline over your Intensity data points and include

the formula on the plot.

Figure 4. Setup required for use of the Solver tool in Microsoft Excel.

The simplest way to quantify error for this process is to calculate an 2 value. Fortunately, the

sum of difference squared is part of the calculation, so only two more equations are needed. The

variance, Var, is equal to the sum of the difference squared values between the observed

intensity values, , and average intensity value, .

2

= ( ) (Equation 6)

2 = 1 (Equation 7)

5

In contrast to 1 relaxation, 2 relaxation is not as simple to determine because it is dependent on

several effects that broaden NMR lines. 2 values are very small for viscous liquids, but

comparable to 1 for low-viscosity liquids. This prevents their use for high-quality spectra

unless additional compounds are added to increase relaxation times. When two neighboring,

identical nuclei have identical precession rates but different magnetic quantum states, they can

exchange these energy states. This causes the lower-energy-state to become excited and the

higher-energy-state to become relaxed. This decreases longitudinal relaxation time and increases

line broadening, and is referred to as 2 . 2 values decrease as molecular motion decreases. 2

can be determined through the Hahn Spin Echo sequence, shown below in Figure 5.

Figure 5. Cartoon depiction of the Hahn Spin Echo sequence used to determine 2 2.

A 2 pulse is applied, and some spins slow down due to local magnetic field inhomogeneities. A

pulse is then applied, which causes slower spins states to go ahead of the main momentum and

the faster spins to lag. Eventually, spin rephasing occurs which eliminates the differences in spin

states. 2 is measured as the spin echo, which causes the dephasing back to the original spread

of phases. Again, we will look change the delay time to look at the intensity changes for the spin

echo. This follows the same exponential curve as Equation 3. However, the inversion recovery

steps are removed for observation, giving the following equation.

= 0 2 = (Equation 8)

By modeling the exponential fit of the graph, 2 is determined. The exponential constant is

related to transverse relaxation in the following way:

1

= (Equation 9)

2

6

The determination of T2 from your collected data will be done in MNova. Detailed instructions

on how to do this can be found here:

http://www.scs.illinois.edu/nmr/handouts/general_pdf/T2Determination.pdf

The data file from the NMR will be emailed to you. You should process the data in this file.

You should follow the instructions on the second page of the document. On Step 3 of the

instructions, you need to select Peak Graph. Remember to copy the data from the generated

chart into your lab report as a table. To access the data go to Advanced > Data Analysis > Show

Table. You should copy the data into excel first and then into word so you can properly format

the table. Be sure to include this in the SI of your report.

The graph will need to be edited. The axes of the graph can be formatted in MNova by right

clicking on it and selecting properties. From there, you can change the horizontal and vertical

axis labels. Please do so to reflect what each axis represents. You can also uncheck the boxes in

the Basic tab under Grid to get rid of the gridlines. Below is an example of what your graph

should generally look like. It is suggested that you attempt to plot your data soon after collection

so you have plenty of time to get help if needed!

trend line from MNova

7

Prelaboratory Assignment

1. Describe the following components of an NMR spectrometer and describe why they are

necessary: super-cooled magnet, shim coils, and sample probe.

2. What is pulse width (pw)? What is pw90 and why is it necessary for determining T1 and

T2?

3. Which relaxation time can be found using an inversion recovery experiment? Which two

pulses are used? Briefly explain the experiment.

4. Why is T2 relaxation often referred to as spin-spin relaxation?

5. How do molecular size and viscosity affect T1 and T2?

Experimental Methods

Equipment:

Quinine/CDCl3 sample

UNITY 400 NB High-Resolution FT NMR Spectrometer

Determine the pw90 for the sample:

Before pw90 is determined, one parameter must be changed to prepare for the future

experiments.

gain = 50 (lowers gain to defined value)

Gather an initial, properly shimmed spectrum. Then, move the parameters and change

experiments using the following:

mp(1,2) moves parameters from experiment 1 to experiment 2

jexp2 joins experiment 2

We will use an array of pulse width values in order to determine the true value.

array a macro that requires the following answers to the prompted questions

Parameter to be arrayed: pw pulse width

Enter number of steps in array: 30 array size

Starting value: 0 (This changes for different probes and different instruments.)

Array increment: 5 T1 values are close

8

These commands set of the remainder of the experiment.

pw[1]=1 replaces first array element, 27, with the value 1.

da displays current array values for pw

d1=10 first delay, given in seconds (allows equilibration)

ai vp=70 sets absolute intensity mode; places spectrum about half-way up on the display.

ga begins acquisition

Repeat the array experiment with lower number of steps and a smaller increment around the

pw360 to get a more accurate result

We can use the following commands to observe the spectra and quit when we have finished.

dssh view accumulated spectra

pw360 occurs when the peak of interest is null.

(NOTE: if the second spectrum is already positive, reset the array with a smaller starting value.)

Finally, we return to the original experiment and set our determined pw90 value.

jexp1 returns to experiment 1

pw90=(the numeric value for the 360 found above)/4 gives pw90 a value

pw=pw90 sets the instrument pw as the determined pw90

Gather an initial, properly shimmed spectrum. Then, move the parameters, change experiments,

and lower the gain using the following:

mp(1,2) moves parameters from experiment 1 to experiment 2

jexp2 joins experiment 2

dot1 sets up the 1 experiment, and prompts for the following:

Enter minimum 1 expected (sec): 5 (a good starting value)

Enter maximum 1 expected: 10 (5 to 10 sec are typical)

9

Enter number of transients (nt): 1 (use minimum nt)

ga begins acquisition

dssh displays completed spectra horizontally

Expand around the signal of interest and set a minimum threshold for the line.

ds(#) displays the last spectrum in the array

dll displays listed line frequencies for the expanded region of interest

fp measures the intensity of each line in the dll for the entire array and outputs the data

t1 executes an exponential curve fitting to determine T1 and outputs the data

printon t1 printoff prints the data into a chart

Write down the observed intensity data. Use these data to create a curve in Microsoft Excel

using the Solver tool in order to determine 1 .

jexp1 returns to experiment 1

First, gather an initial, properly shimmed spectrum. Then, move the parameters and change

experiments using the following:

mp(1,2) moves parameters from experiment 1 to experiment 2

jexp2 joins experiment 2

doT2 loads the pulse sequence for a 2 experiment

10

T2setup sets up the 2 experiment, and prompts for the following:

Enter minimum 2 expected (sec): 1 (0.1 1 sec is good to start)

Enter maximum 2 expected: 8 (5 to 10 sec are typical)

Enter experiment time (hr): 0.2 (will be slightly different than actual time)

ga begins acquisition

Expand around the signal of interest and set a minimum threshold for the line.

ds(#) displays the last spectrum in the array

dll displays listed line frequencies for the expanded region of interest

fp measures the intensity of each line in the dll for the entire array and outputs the data

t2 executes an exponential curve fitting to determine T2 and outputs the data

printon t2 printoff prints data into a chart

Write down the observed intensity data. Use these data to create an exponential curve in

Microsoft Excel to determine 2 .

jexp1 returns to experiment 1

Tables: include the observed time and intensity values used to determine both relaxation time

constants. These should make up the two tables that you need in the body of the report.

Simply report pw90. Make sure that you tabulate all the data that you use in producing your

graphs.

Plots: use Microsoft Excel for the T1 plot and MNova for the T2 plot. Be sure to label axes

and title the graphs in a descriptive manner. The graphs should be easy to read. You should

have two plots: observed intensity vs time for both time constants. Plot the formula values

from solver over the observed intensity as a black trendline on the T1 graph and display the

formula with the correct constants on the plot using a text box, as well as the calculated R2

value. For the T2 graph use a text box to display the exponential trend line on the graph. You

dont need to modify the MNova graph to make it look like the excel graph but be sure you

have all of the required labels.

11

Calculations: show how you calculated the following,

pw90

1

2

Attach your original copy of your lab notebook in the supplemental information (SI) section

of your report. Also in the SI include the table from Excel Solver that you used to obtain T1

and the Table from MNova that you used to obtain T2.

For the discussion section, make sure you include answers to the questions below, doing so

in a discussion format (i.e. DO NOT put Question 1, then Answer 1, etc.). This is required

in addition to normal discussion procedures. The idea is not to only answer the questions

below but answer them as support for your normal discussion. The chemical engineering

question should be placed according to the lab format guidelines which can be found on

Compass. Warning: Be sure to cite ALL of your sources appropriately. Including

information that comes from an outside source and is not cited will result in point deductions.

Questions

1. What is your pw90 and what effect would it have on the data collected if it were different?

2. What are your relaxation constants? Based on the properties of Quinine, are they what you

would expect?

3. You used two different software programs to process the acquired NMR data. Which gave you

a better understanding of the trends in the data? Why? (There is not only one right answer to

this question, I want you to think critically about the data processing methods used.)

4. Quantify sources of error. What are some common sources of error in NMR spectroscopy in

general?

1. NMR spectroscopy, though a very useful technique, is plagued with many issues such as

high costs and low sensitivity. Despite these drawbacks, NMR spectroscopy is frequently

12

used in many areas of chemistry. NMR is also used in industry. Discuss an example of an

industrial application of NMR spectroscopy that stands out as particularly interesting

and/or useful to you.

13

## Viel mehr als nur Dokumente.

Entdecken, was Scribd alles zu bieten hat, inklusive Bücher und Hörbücher von großen Verlagen.

Jederzeit kündbar.