Proton (7H) NMR finds use for both qualitative analyses and quantitative analyses; in this section we briefly consider each of these areas. Identification of Compounds Proton NMR is an essential tool for the qualitative analysis of organic, inorganic, and biochemical compounds. Figure 19.4.119.4.1 provides a simple example that shows the relationship between structure and a "H NMB's peaks. The spectra in this figure are for a set of four simple organic molecules, each of which has a chain of three carbons and an oxygen: 1-propanol, CHCH:CH:0H, 2-propanol, ‘CH3CH(OH)CHs, propanal, CHsCH2CHO, and propanoic acid, CH3CH2COOH. The first two of these molecules are alcohols, the third is an aldehyde, and the last is an acid. The main spectrum runs from 0-14 ppm, with insets showing the spectra over a narrower range of 0-5 ppm. Each of these molecules has a terminal -CHs group that is the most upfield peak in its spectrum, appearing between 0.94 ~ 1.20 ppm. Each of these molecules has a hydrogen that either is bonded to an oxygen or a hydrogen bonded to the same carbon as the oxygen. The hydrogens in the ~OH groups of the two alcohols have similar shifts of 2.16 ppm and 2.26 ppm, but the aldehyde hydrogen in the -CHO group and the acid hydrogen in -COOH are shifted further downfield appearing at 9.793 ppm and 11.73 ppm, respectively. The hydrogens in the two ~CH.~ groups of 1-propanol have very different shifts, with the one adjacent to the -OH group appearing more downfield at 3.582 ppm than the one next to the ~CHs group at 1.57 ppm. Not surprisingly, the -CH~ hydrogen in 2-proponal, which is adjacent to the -OH group appears at 4.008 ppm, Comparisons such of this make it possible to build tables of chemical shifts—see Table 19.2.1 in Chapter 19.2 for an example—that can help in determining the identify of the molecule giving rise to a particular NMR spectrum. As this receives extensive coverage in other courses, particularly courses in organic chemistry, we will not provide a more extensive coverage here.Figure 19.4.119.4.1.H NMR spectra for 1-propanol, 2-propanol, propanal, and propanoic acid, The full spectrum are shown using the scale at the bottom of each figure. The insets show close-ups of the NMIR spectra from C— 5 PPM. The original data used to construct these spectra are found here. The spectrum for propanal sar recorded on a 300 MH2 instrument; the other three spectra were recorded ‘ona 90 MHz instrument. Quantitative Analysis ‘quantitative analysis requires a method of standardization, which for NM ovally makes use of an internal standard, A good internal standard should have tigh purty and should have a relatively simple NMR spectrum with peaks that do rot overlap with the analyte or other species present in the sample. f we are jaterested in only the relative concentrations of the analyte and the internal standard, then we can use the following formula Me ta Mu Mu Ta No where M is the molar concentration of the analyte or internal standard, | is the intensity of the NMR peak for the analyte or internal standard, and Nis the ‘number of nuclei giving rise to the NMR peak for the analyte and the internal standard, Even if we don't know the exact concentration of the internal standard, if we know that its concentration is the same in all samples, then we can determine the relative concentration of analyte in a colle of samples.lf we are interested in determining the absolute concentration of analyte in a sample, then we must know the absolute concentration of the internal standared; when true, then Equation 19.4.119.4.1 becomes M Ww _ pate Nin, Ma Wn yp 0= 7° NM Wa where W is the weight of the internal standard or the sample that contains our analyte,‘Te cugg conan nepenen ct magne fe stengh bcos sae byte magnets fll of ante ces, rote Spero ragnet Threte, qustes ne reqieny) art pm cna st) inane mae a rn sna a2 5 oman prtan we ak te pit vo by te psn a ppm. Becaue ne magn of iisaconis te rae he ping walshave re sae coil casi Mz par The pec woul ave wo sore each erg 3. doublet. {rte cit poke pn a cote ye Ch paw. on peak m+ 98HZ ane al pom 3 SHE sping ox ouPing In consequence the CH peak at 2.5 ppm wl be spl twice by each proton from the CH. The fist proton wil split the peak info two equal infensites and wil go from one peek at 2.5 ppm to two peaks, one at 2.5 ppm + 3.5 Hz and the other st 2.5 pom - 3.5 H2—each having equal intensities. However these will be spit again by the second proton. The frequencies wil change accordingly: + The25 ppm + 3.5 He signal wil be split ino 2.5 ppm +7 Hz and 2.5 ppm The28 ppm. S.6He eignal willbe epitinto 26 ppm and 2.6 ppm 7 He ‘The net results nat a signal consisting of 4 peaks but three: one signal at 7 Hz above 2.6 ppm, wo signals occur at 25 ppm, and a final one at 7 Hz below 2.5 ppm. The ratio of height between them is ‘12:1, This is known as a triplet and isan indicator thatthe protons three-bonds from a CH group, ‘This can be extended to any CH, group, When the CH.-CH group is changed to CH.-CH., keeping | the chemical eit and coupling constants identical, the folowing changes ere observed 1+ The relative araas between the CH, and CH, subunits will be 3:2 | “The CH coupled to two protons into @1:2:1 triplet around 1 ppm. ‘+The CH. is coupled to three protons Something spit by three identical protons takes a shape known as a quartet, each peak having relatve intensies of 13:3:1 peaks spit nKdencal protons inte compenens whose sizes aren the rao of the th row of Pascale tanglea Name Row 0 singlet 1 1 doublet 1" 2 Wiplet 124 3 quartet 1334 4 quintes 14644 5 sotet 15101081 7 octet 172135352171 8 nonet 1828567056281 Because the ri ow has 1 components his type of epiting Is suid to flow the net proton with n neighbors appears as a cluster of nei peake hn Z-methypropane, (CH,),CH, as anaher example: the CH pots is atached to twee dered ‘methy! groups containing a total 9 identical protons. The CH sia! in te spectrum would be spit no ten peaks according tothe (n+ 1) fle of Muli. Belo are AM agra cero ase several simple mules of ts type: Not that he outer ines ofthe not (arch ceri ne hah as those of te Second peak can baraly bo Seen, Ging spatial eammisear yy eee I | AM sll. LLL ni scan coupled to two ferent protons then the coupling constants ae ely to ‘ferent and instead ofa tipot omen? Of doublets wil be seen, Siiay. a protons ecopledto two other protons of one type, and a thd of another type with a itferont, smaller coupling constant, then a triplet of doublets is seen. In the example below, the triplet coupling constant is larger than the doublet one. By convention the pattern created by the largest coupling constant is incicated frst and the splitting pattems of smaller constants are named in tur. In the casa below i would be erroneous to refer to the quartet of triplets as a triplet of quartets. The analysis of such multiplets (which can be much more complicated than the ones shown here) provides important clues fo the structure of the molecule being studied | | SW a oe ll Doublet of doublets Triplet of doublets Quartet of tips ii } Wh, Wy ‘The simple rules for the spin-spin spliting of NMR signals described above apply only if the chemical shifts of the coupling partners are substantially larger than the coupling constant between them. Otherwise there may be more peaks, and the intensities of the individual peaks will be distorted (second-order effects).Ethyl acetate H Ho on | | H-¢--0-¢-¢-H | ' HH | Chemical shift (ppm) Example 1H NMR spectrum (1-dimensional) of ethy te plotted as cignat intensity vs. chemical shift. There are three different types of H atoms in ethyl acetate regarding NMR. The hydrogens (H) on the CHsCOO- (acetate) group are not coupling with the other H atoms and appear as a singlet. but the GHz and -CHs hydrogens of the ethy! group (CHzCHs) are coupling with each other, resulting in a quartet and tnplet respectivelyLUMEN: MCC Organic Chemistry Application of Proton NMR OBJECTIVE ‘After completing this section, you should be able to use data from 'H NMR spectra to distinguish between two (or more) possible s:ructures for an unknown organic compound. There will be cases in which you already know what the structure might be. In these cases: + You should draw attention to pieces of data that most strongly support your expected structure. This approach will demonstrate evaluative understanding of the data; that means you can look at data and decide what pails are more crucial than others. + You should also draw attention to negative results: that is, peaks that might be there if this spectrum matched another, possible structure, but that are in fact missing, One of the most complicated problems to deal with is tre analysis of a mixture. This situation is not uncommon when students run reactions in lab and analyse the data. Sometimes the spectra show a little starting material mixed in with the product. + Sometimes solvents show up in the spectrum. ‘As you might expect, the minor component usually shows up as ‘smaller peaks in the spectrum. If there are fewer molecules present, then there are usually fewer protons to absorb in the spectrum. 'n this case, you should probably make two completely separate sets of data tables for your anal lysis, one for each compound, forthe main compound and one for impurities," T° OnRemember that integration ratios are really only meaningful within 2 single compound. if your NMR sample contains some benzene (CeH) and son acetone (CH:COCHS), and there is a peak at 7.15 thal integrates to 1 proton and a peak at 2.10 ppm integrating to 6 protons, it might mean there are 6 protons in acetone and 1 in benzene, but you can tell that isn't true by looking at the structure. There must be six times as many acetone molecules as benzene molecules in the sample. ‘There are six protons in the benzene, and they should all show up near 7 ppm. There are six protons in acetone, and they should all show up near 2 ppm. Assuming that small integral of 1H for the benzene is really ‘supposed to be 6H, then the large integral of 6H for the acetone must also represent six times as many hydrogens, too. It would be 36 H. There are only six hydrogens in acetone, so it must represent six limes as many acetone molecules as there are benzenes. Similarly, if you have decided that you can identify two sets of peaks in the 'H spectrum, analysing them in diferent tables makes it easy to keep the integration analysis completely separate too ; 1 H in one table will not be the same size integral as 1 H in the other table unless the concentrations of the two compounds in the sample are the same. However, comparing the ratio of two integrals for two different compounds can give you the ratio of the two compounds in solution, just as we could determine the ratio of benzene to acetone in the mixturs described above We will look at two examples of sample mixtures that could arise in lab. Results like these are pretty common events in the labin the first ‘example, a student tried to carry out the following react on, a borohydride reduction of an aldehyde. The borohydride should give a hydride anion to the C=O carbon; washing with water should then supply a proton to the oxygen, giving an alcohol.From this data, she produced the table below. it leugasin kn) maine maapicty pat sbuctte assignment i, "opm tm tage ® & B8pom zm wk aH © 8156 omen aH ay 420m 0mm Moa © 780m 9mm mh ay 1 83pm 2m H 9 4750 sme may ee te ae Vane sem sem ge) : ans et SIRO gg 262 een pa 47 om 4/29 een TENSES SES OMENS preteen pom 4/2993 mane TOMER 2185. sete some 4 rasa pan Fees eiThey are toth 2H in her those two peaks are equal, pee oe she notes that within each ee i ee the second represents 2H. at mean’ a 7 Sees 4 as there are molecule 2. That way, there many of , wwouls be ee Gri=0, and its integral would be the same as the 1 x CH2-0 in the other molecule. ‘One way to approach this kind of problem is to: |. choose one peak from each of the two compounds you want fo compare, | decide how many hydrogens each peak is supposed to represent in a molecule. Is it supposed to be a CHe, a CH, a CHs? «divide the integral value for that peak by that number of hydrogens it is ‘supposed to represent in a molecule. «compare the two answers (integral A / ideal # H) vs (integral B / ideal #4). «the ratio of those two answers is the ratio of the two molecules in the sample. So there is twice as much aldehyde as alcohol in the mixture. In terms of these two compounds alone, she has 33% alcohol and 56% aldehyde. That's ( 1/(1+2) ) x100% for the alcohol, and ( 2/(1+2) ) x100% forthe aldehyde. That calculation just represents the amount of individual ‘component divided by the total of the components she wants to compare. ‘There are a number of things to take note of here. Her reaction realy didnt work very wll. She stil has major material, not product. peat eels) She will get a good grade on this lab. Although the ex; ‘periment didn't work well, she has good data, and she has analyzed it very clearly, She has Separated her data table into cnt diferent sections for cron Sometretht mate oaser anaes tage + She has noted the Shs ce ne ct egal data he may have measured he ruler) and also Converted ino arth 2 more convenient sed onthe niga fora peak thal she fall colar aon«She went one step further, and indicated the internal integration rat within each individual compound. i ion usit integral data She calculated the % completion of the reaction using the in for the reactant and product, and she made clear what part of the data she used for that calculation, A similar procedure could be done if a student were just trying to separate two components in a mixture rather than carry out a reaction. She also calculated the overall purity of the mixture, including a solvent impurity that she failed to remove. + However, CHCls is not included in her analysis of purity. CHCl really isn’t part of her sample; it was just present in the NMR solvent, so it doesn't represent anything in the material she ended up with at the end of lab. Another student carried out a similar reaction, shown below. He also finished the reaction by washing with water, but because methanol is soluble in water, he had to extract his product out of the water. He chose to use dichloromethane for that purpose. aes oro o on He obtained the following data.fa, he constructed the following table From this dat en peuioncee met ‘There are some things to learn about this table, too. + Does the integration ratio really match the integral data? Oris this just wishful thinking? ‘This table might reflect what he wants to see in the data, But what else could be in the data? + CHC! is often seen in NMR spectra if CDCl is used for the NMR sample, It’s there, at 7.2 ppm. + “Leftover or residual solvent is vary commen in real lab data. There it is, CHeCle from the extraction, at 5.4 ppm. + What about water? Sometimes people don't dry their solutions property before evaporating the solvent. There is probably water around 1.5 to 1.6 ppm here. This student might not get a very good grade; the sample does not even show up in the spectrum, so he lost it somewhere. But his analysis is also oor, so he will really get a terrible grade. EXAMPLE Three students performed a synthesis of a fra ts p nthesis of a fragrant es SiNCHCO:CH:CHs During their reactions, they each used a eiffecnt sober Th a ons were ofle (© 82° peaks in the NUR spectrum for To make their NMR sortak® OF Cheroform (CHCI, in the CDChs they + See the first student's spectrum, See the second student's spectrum, ster, ethyl propanoate,+ See the third student's spectrum. 1 in ‘They were also able to determine that they had some ltover solar their samples by consulting a useful table of solvent impure {which ty found in Goldberg etal, Organometalics 2010, 29, 2176- 2179) 1. What is the ratio of leftover solvent to ethyl propanoate in each sample? 2. What is the percent of each sample that is leftover solvent Additional NMR Examples For each molecule, predict the number of signals in the 'H-NMR and the “C-NMR spectra (do not count split peaks - eg. a quartet counts as only one signal). Assume that diastereotopic groups are non-equivalent °1 the number of predict the numb common amino acids, trum, wupled "C-NMR spe 5.2: For each of the 2 ps casa he ern NN cout tna eto, D9 ST sm obtained on a 300 MHz instrument obtained on a 100 MHz instrument P54: Co Miz instrument. The ch ne easily recognizable splitting pattern for the eromatic proton P5.5: Or ras from disubstituted benzene structures is a pair of doublets. Does is pattorn indicate ortho, meta, or para substitution? P5.6 :Match spectra below to their corresponding structures A-F,Spectrum 4 8 sping 4a ‘ 245 t a Quintet tar t Spectrum 2 : splitting 368 s 290 : vo quintet Spectrum 3 integration 2 2 1ata 3.22 427 4.13 Spectrum 5 6 4.18 1.92 1.23 ost ‘Spectrum 6 6 3.69 2.63 splitting splitting splitting q q t t splitting s integration 1 1 15 integration integration integration 15 1PS.7: Match spectra 7-12 below to their coresponding structures Gl ‘Structures: ‘Spectrum 8: 9.36 655 splitting (0-0 genial ving ress) integration 1 126 1.99 0.96 Spectrum 9 8 957 6.30 6.00 1.84 Spectrum 10: 3S 9.83 227 4.07 ‘Spectrum 11: 975 2.30 221 0.98 splitting s 8 8 s splitting t 4 splitting t cr integration 1 1 1 3 integration 1 2 9 integration 1 2 1integratio 1 een below to their corresponding Ho, 0 Ht N O. © M ° ° l ° ° on Oj a Cy 2 Ae | P Q R ' ‘Spectrum 13; 3 splitting integration 8.15 7 1633 d Spectrum 14: 1-723C (structure O) 8 6.05 224 ‘Spectrum 15: 857 7.89 6.30 2.28 splitting s s spliting s(b) d a s splitting s(b) s s s(b) integration 1 3 integration 1 integration 1 1 15 splitting 4.03 s 2.51 t 2.02 t 5.9: Match the ‘H-NMR spectra 19-24 below to their corroay structures S-X, Structures’ integration 1 1 1 spondingv Spectrum 19: 8 9.94 717 7.31 2.43 ‘Spectrum 20: 8 10.14 8.38 87 775 a splitting splitting s 4 integration 1 enn integration 2 1 22.42 ‘Spectrum 23: 7.40 6.86 3.78 3.61 242 Spectrum 24: splitting integratio integratic 25 15 integration 15 15 integratic7.23-7.30 3.53 P5.10: Match the 'H-NMR spectra 25-30 below to their corresponding structures AA-FF. Structures: Ph. ° Ph AA prot doesnot O;N, Stow ald poten). = N HOY CHs ° DD Spectrum, 5 splitting 9.96 s 779 d 7.33 d 272 4 1.24 EE FF integration 1 2 2 2 3integrat 2 2 6 integrati 10 fl 3 3 splitting integration 8.08 s 1 7.29 d 2 6.87 d 2 5.1 5 . 2 3.78 s 3 ‘Spectrum 29: 3 splitting integration718 d 1 6.65 m 15 32 q 2 1.13 t 3 ‘Spectrum 30: 8 splitting integration 832 s 1 4.19 t 2 2.83 t 2 2.40 s 3 5.11: Match the ‘H-NMR spectra 31-36 below to their corresponding structures GG-LL sucuss n ve tr ue ° 2 “OL Ch hoe I . t u ce aH un F672 653 481 3.18 224 1.22 ‘Spectrum 33: 7.08 splitting d integration 2671 6.54 3.69 3.54 ‘Spectrum 34: 9.63 7.45 677 3.95 2.05 1.33 ‘Spectrum 35: 9.49 7.20 6.49 482 1.963 ‘Spectrum 36: splitting splitting integration eennn integration 1 2 2 2 3splitting d(d=t Hz} d(d-t Hz) q integration 1 1151.77 132.99 20.91 11.92 'b) Molecular formula: CrH14O2 JHENMI 5 splitting 3.85 a 2.32 q 1.93 m 1.14 t 0.94 a CHe CHe CHa integration 2 2 1 DEPT CHe cH CHe CHs CHs32.61 26.04 4) Molecular formula: CicHy20 ‘H-NMR: 5 splitting 7187.35 m 3.66 s 2.44 q 1.01 , CHe CHs DEPT integratic 25 1 fl 18208.79 134.43 129.31 128.61 126.86 49.77 35.16 175 5.13: a) 461.12 65.54 21.98 10.381 b) cH cH cH CHe CHe CHe IMR. data ie given for the molecules shown below. Complete the peak Iment column of each NMR data table. carbon #171.76 60.87 58.36 24.66 14.14 8.35 carbon #d) 173.45 155.01 130.34 125.34 115.56 2.27 40.27 e) 147.79 129.18 115.36 CH CH carbon # carbon #111.89, CH 44.29 cH. 12577 CH: 5:14: You oblain the following data for an unknown sample. Deduce its structure, HNMR:Prue) Mass Spectrometry: a- mz P5.15:You take a 'H-NMR spectrum of a sample that comes from a bottle of 1-bromopropane. However, you suspect that the bottle might be contaminated with 2-bromopropane. The NMR spectrum shows the following peaks: 5 splitting integration 43 septet 0.0735 34 triplet 0.6610.685 0.441 1.00 fically, what percent of the c this chapter include a signal due Explain the spitting pattern for ° ye aot H, HOH aa) 7 (ee NN cH Ny ne? NA 1 a » BUM the products 2A and 28 have identical or diferent "H-NMR spectra? Explain, b) Suggest a "H-NMR experiment that co ud be used to determine what Percent of starting material (1) got turned into product (2A and 28), ©) With purified 2/28, the researchers carried shown below to make 3A, and 3B, known as ‘M 38 have identical or different "H-NMR spectra’ ‘ut the subsequent reaction. losher’s esters’. Do 3A and ? Explain4d) Explain, very specifically, how the researchers could use ‘H-NMR to determine the relative amounts of 2A and 2B formed in the reaction catalyzed by yeast enzyme. es EXERCISE Question How can H' NMR determine products? For example, how can you tell the difference between the products of this reaction? co Hel a cor SOLUTION ae Show AnswerYes, you are able to determine the difference in the spectra. For the 2- chloro compound will have multiple quartets while the 1-chloro compound will only have a quintet and a triplet for the signals in the ring: