CHM 319L Organic Laboratory, Dr. Laurie S. Starkey Introduction to Mass Spectrometry Mass spectrometry is used to determine a sample's molecular mass and molecular formula, Some structural information can also be determined by mass spec. This teclinique is especially valuable when used in conjuction with gas chromatography (called GC-MS); the GC separates the components of a mixture and then the mass spec analyzes cach component. To obtain a mass spectrum, a sample is vaporized and then bombarded with a high energy beam of electrons (this technique is called electron-impact ionization, or ED). ‘This ionizes the sample by knocking an electron out of the structure. A nonbonded or bonded electron is removed, resulting in radical cation species. Although it is now missing an electron, the intact molecule still has the same molecular mass and is known as the molecular ion (M*), or the parent ion. In addition, this high-energy environment causes the molecular ion to fragment and the various pieces can be analyzed to learn something about the original structure. All charged species are separated by mass as they are passed through a magnetic field and a detector records how many pieces of each mass have been formed. Note that because this technique does not involve the absorption or emission of energy, it is not called spectroscopy. blast with & electron beam fragmentation occurs magnetic n defle — 7 separate ‘charged ® [ie icles im mass-to-charge sample of a e loss of an mixture of charged ratio, m/z neutral molecule electron results in and uncharged (charge is is vaporized aradical cation particles usually +1) (no change in mass) detector records the frequency of @ > & each mass; the taller the peak, the more abundant the fragment lower mass higher mass os MASS SPECTRUM Qxkret 2 eaeer) eee base peak (most abundant) defined as 100% Spectral Analysis In this spectrum, the sample has a molecular mass (M) of 100 amu. 80- molecular ion, M* S to set scale : The most abundant a 60- Stathe| a anteat (highest m/z) fragment, called the base 5s (rag >| gives molecular peak, has a m/z of 43 a mass (in amu) amu, or (M-57). This 3 40- fragment was formed é when a 57 amu segment of the molecule broke Ps off. Many other fragments were formed; not all will be analyzed. 0.0 20 40 60 80 100" 120 mz noe that a small peak is observed spectra have been reproduced from the NIST Chemistry cat (M1) 7 WebBook hitp:/rebbook.nist gov/chemistry 7What is the mass of a single molecule? e cH, Mass= C 44g) = 1% + A0)= Ty om (MN) but ~1% of carbon atoms exist as 3C isotope! Mas= "CO 44gy = 13+ 40) = [7 ae Cnet) peat = mie a M* peak What ratio is expected for the molecular ion peaks of C)H,? ta CaeClts wht on CHLCH, acma (mel) » 2% A loo 1 ' the number of carbon atoms affects the relative height of the M+1 peak CH, Other isotopes of high abundance C1545 ann) atoms are 76% 25Cl and 24M BIL Mee : me Br (79.90 amu) atoms ate 51% "Brand 40% 8tpy ow (3 WS MA 4 fragments containing Cl or Br have significant M+2 peaks ‘What ratios are expected for the molecular ion peaks of CHpBr,? m2 ia im 7 possible masses? CH, Br Br total amu | sce \ WW 7474 2 Ltd for gi 74 tae 14 Wezel cree p94, in cael wy ut Bh ; Sry Beaiaa] , u ¥ Obi. 2 « ey te n a i ) 7 é IR sb eras aaa i mass spec of chloroethane (M* 64) mass spec of broniobutane (M* 137) mass spec of dibrorimethane (M* 172) Determining molecular formula from high-resolution mass spectrometry (high-res mass spec) exact masses of various elements: CyHgN2O 98.0480 amu "1.007825 ama ea these molecules all CsH6O, 98.0368 amu "°c 12,000000 amu have a molecular etic pr reas '4N 14.003050 amu mass of 98 glmot pete 3 CgHjpO 98.0732 amu 0 15.994914 amu CH aren the molecular formula ean be determined from precise molecular mass LMass Spee Features of Various Functional Groups Fragmentation of the molecular ion Which is more stable, a carbocation C* Hee ben om ys. ora radical Re? coe e- yp mere MWe Was st (clever 0 ecto?) fragmentation is more likely if it gives more stable carbocations and radicals Be> 2 7 ly rytthoyl pid ms CH3—CH,—CH,—CH,—CH,—CH,; o> [CHe— ‘CH)—CH)—CH,—CH, =CHs | hexane ———— — iaament | te~CHe= CH= CH,-CHy : |. fragment (CHs—CH,—CH,—CH,-+.CH,—CH fF inde 57 (M29) a break a bond to give two fragments: ‘one cation and one radical CHy—CH—CH>—CH, mle 29 (MST) 4, fragment a CHp~CHp—CH, |* —> CH3—CH)—CH, ¢ (CHy—CH,—CH CH)—CH,—CHg mlz 43 (M-43) voi) int JH ‘Cry—ct OHO CH2—CH»—CHp oH, Ge ragment CHy—CHp—CHp—CHy— ct-+ors |! pid mi TL (MEAS) ad + * CHg—CHy—CH>—CHy—CHp CH m/z 15 (M-71) note: hexane has peaks at m/z15, 29,43, 57, 71, 86 ei ke Branched Alkanes 204 ‘S Explain why the base peak of 2-methythexane is at m/z43 (M57). yc oo Be He : Rs Cly—CHy—CHy— CH} CH-CHy 20- t Ld | cnt Tay 00 Es 4 & 80 100 wie Why are the peaks at m/z 15 and 71 so small? ‘ by—clh, * Urs Lily crynontth Che Map tna sable Wnt mde ane 3Alkenes remove . " OHy~CH~CHp—CH=CH—CH, 12> [CHy—CH»—CHp—CH=CH—CH,|* allylic cleavage gives stable carbaeation Nyrt beat bowed en E fragment bp Ln CH,—CH,—+-CH,—CI ra H—CH, YOY + a CH,—CH,4CH,—-CH—CH—CH, | CHa CHe 2, high-energy pi electron —_—> OacH ie is most likely removed i Bae allylic carbocation is resonance-stabilized Aromatic ie bie 241 oncontba {> : =e Beh) eS C be Cly \umyyle a ee. be FEAL OAS O wopylium cation mz91 Alcohols 2 D : cleavage | x (, [eHs—cH,-tcH,—on]" SSR | Gy-GH <> Ch Zvi yea stabbed CO ‘OH CHs—CH,~“CH>~- high-energy nonbonded electron is most likely removed 3kalso, loss of HyO (M-18) may be observed (multistep mechanism first involves an intramolecular proton transfer)‘O-CH,—CHy—R! < Ethers a [Roti fcr,—0-Cr$ cher] a. 4 R—CH,—CH,—O~CH, also, loss of RO+ may be observed “ + Yo OReleavage oy CHa di Cu -oe) + [ory —cHp—cHe ton] eal a 0k -45 che Amines arcleavage : ) ® [ors —cre korea! enti be & chen | feomance 2}.Nitrogen Rule: M* is odd if there are an odd number of nitrogens! odd molecular mass indicates one nitrogen (or 3 or S or 7 nitrogens...) even molecular mass indicates zero nitrogens (or 2 or 4 or 6 nitrogens...) Aldehydes & Ketones e.. Go 0: i Ww \ R. +C6—R mR mlz (M-R) + acleavage acylium ion is resonance-stabilized loss of larger group more likely (more stable R+ } McLafferty rearrangement is a common fragmentation for carbonyl-containing molecules es + ka aH el MeLafferty ZH . HR oF GouA rearrangement a aie SEEDRESuEEigl Cx Roo, CHe i CL NUCH. REET | vorable membered ; + ring in transition state = i‘ clam ad \ rake molecular ion cleaves li ebgereed between cand B carbons Esters Oo esters can undergo a cleavage, McLafferty i rearrangement, or loss of RO r4-ctorYue 25- 30 mune ee poy ens Spectrometry Diseyssion Questions ae For the given molecule (M=8%4), do you expect the more abundant peak to be m/z 15 omiz 433 Explain. o : 2 ob CHeH ch “ey & 5 Becher | bth For the given molecule (M=74), which peak do you exps - to be most abundangé m/z 21 45 or m/z 592 Explain, 2 el Son, o- ae HO-CH,4CHy CHS CH, —~ fig iy 2 Ke (43) Explain why the mass spectra of methyl ketones typically have a peak at m/z 43, Provide the structure of this fragment. oe Cy eee Cho We WS CHy$-CHp—CHp—CHs -24 -\S In the mass spectrum of the given molecule (M=88), account for the peaks at m/z 45 and m/z 57. eons a - Bl CHy-O#CH4CH,—CH»—CHy Fach 4257 Cn ) ae & cto Se mle 4s (m-43) How could you use mass spectrometry to distinguish between the following two compounds (M=73)? Provide strictures (and m/z values) for the significant fragments expected CHy—CHy- CHE CH NH and CCH Loty—NH— Chis ao wh By NA b 32 wh 1 -\>d e m2 wy > “677 Cc What would be the m/z ratio forthe fragment resulting from a McLaffesty sage for the following molecule (M=4469? What fragment accounts for its base peak at m/z aff (in - _ Ae sa tS tS aby Fat las ches ~57 — = 72 Clty? “ts ue | : 7 oc Wade CAIRG Ba tO Wr]. y [sgt con