C8953 NMR strukturní analýza seminář Identification of an unknown compound Jan Novotný 17 6003@is.muni.cz April 1, 2020 Task 0: Classification of an unknown substance jlL*1_ A k A/UlA A 1 V a /A aA*.. . , I JAJl JÜU_UlJ_L -i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—r T 5 Ondřej Jurček JL-IL. i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—r~ 1 á i fiP >2 4 4 11 i W= >0 o m^rV 26, 2S7 ^)3V(^ Task 0: Classification of an unknown substance Terpene Carbohydrate Streroid Peptide a í AAaaA Alkaloid 1 Jul Coumarine I II 111 I Jk_ U -i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—r r 3 2 1 ppm 3|= Ondrej Jurček m>n 26, 2M7 03V<^ Task 1: Ci0Hi3NO 5[ppm] Multiplicity Integral 1.05 triplet 3 1.75 singlet 3 3.70 quartet 2 7-7.60 complex multiplet 5 Ondřej Jurček m>rV 26, 2m7 0*V<^ Task 1: Ci0Hi3NO 5[ppm] Multiplicity Integral 1.05 a triplet 3 f A-1\| 1.75 c singlet 3 \,_// \_ 3.70 b quartet 2 e d // 7-7.60 d-f complex multiplet 5 O C / Ondřej Jurček m^rV 26, 2fL7 "O^V^ Task 2: CnHi5N02 5[ppm] Multiplicity -/(Hz) Integral 1.30 triplet 7 3 3.00 singlet - 6 4.25 quartet 7 2 6.65 dublet 8 2 7.80 dublet 8 2 26, 2m7 050/(9^ Task 2: CnHi5N02 5[ppm] Multiplicity J(Hz) Integral ^ -^ O 1.30 b triplet 7 3 N 3.00 e singlet - 6 4.25 a quartet 7 2 6.65 d dublet 8 2 7.80 c dublet 8 2 Ondřej Jurček T^ri' 26, 2fL7 ^3V<^ Task 3: Ci2Hi7NO - ^/COSY JH NMR (300 MHz, CDCI3) i>«ii'i,»iiimLi>«»J»» Ondřej Jurček F2 (ppm): 1 I I I I I MM MM--MM MM--MM I I I I--lili I I I I--1 I I I MM--1 TT Mil--1 I I I n-i--1 i ! i 140 120 100 80 60 40 20 0 Fl (ppm) m^rV 26, 2fL7 ^07Q/(^ Task 3: Ci2Hi7NO - ^-"C/HSQC.HMBC HMBC (300 MHz, CDCI3) - F2 (ppm) 1- 2 3" I | I I I I | 1 I I I I 1 I I | I I I I I I I I | I I I I I I 1 I | I I I I I I I I | I I I I I I I 1 | I I I I II I I | I I I I I I I I [ I 1 I I I I II | I I I I I I I I | I I I I 180 160 140 120 100 80 60 40 20 Ondřej Jurček m^rV 26, 2fL7 ^07Q/(^ XH NMR (300 MHz, CDCI3) I 111 1111 111111 1111 111 111 111 1111 | í I 111 I H I | M 111 11! I | í | fll ll| lili llllj II ll|MII niljll lljll II jllll IIII |llll| 7.7 7.5 7.3 7.1 ppm 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 ppm 0.92 1.08 1.84 1.00 2.08 2.05 2.91 2.94 2.02 COSY(300 MHz, CDCI3) F2 (ppm): I" T--1 I I 1 i i i--r—r L_L-Jul i si í ' 1 ' 1 I 9876543210 Fl (ppm) Ondřej Jurček m m^rV 26, 2fL7 ^ypj®' General comments • inspect molecular formula CmHh0oNnXx: Degree of unsaturation m + 1 — 0.5(h + x — n) 9 identify signals of CH3 and exchangeable protons in ID 1H sectrum 9 arbitrary numbering (e.g., from lower to higher value of chemical shift) of resolved resonances in all spectra • identification of the individual spin systems using DQF-COSY • resolve geminal protons using HSQC • connect molecular fragments/isolated spins using HMBC, NOESY specify the stereochemistry (relative configuration) by means of J— and NOE interaction • in ID spectrum bottom blue numbers are integrals, labels in violet frames contains the arbitrary label (A-N), multiplet specification (use with caution, automatically determined), and position of a signal in ppm 9 UnHa-UnHb in 2D reffers to correlation of protons a and b of unknown compound Un 010/0^ ID *H of CioH200 4.0 3.5 3.0 2.5 fl (ppm) 2.0 1.5 1.0 0.5 Jan Novotný < a t3 - - 1H-13C HSQC and NOESY 2CH UnC1-Ha UnC2-Hd > UnC3-Hqs^^ 30i E ü 50i 60i 70i UnC4 _ UnC5-HE— j UnC6-HhL UnC4-Hf UnC7-Hj \ i UnC7-Hb UnC8-Hk UnC8-Hc / UnC9-Hg UnC10-Hm \ 20 30 40 50 60 70 H E Ä 2 3i 4i 4 2 C02-1H (ppm) 6 _i_ uhHa-Hn 4 —i— _2 UnHd UnHk-Hr hA UnHd-Hk UnHe-Hm llnHa Hm Unŕla"HI f UnHb-Hj , m UnHa-Hm i f _ UnHb-^i UnHa-Hm i / UnHd-H^l UnHg-Hn UnHf-H UnHg y UnHf-Hj ^UnHf^ UnHh-Hm / ^ UnHh-Hk Bi © - chiral carbon 9 E a 2 4 _i_ UnHg-Hm -1- 4 UnHb-Hh UnHb-Hj I UpHb-Hf UnHc-Hk \ UnHb-Hi| 3 2 Q)2- 1H (ppm) Jan Novotný >0 0,0 4/5 Task 1: J-con necti vity of C10H20O 5 4 3 2 1 Task 1: J-connectivity of C10H20O • CH 9g has crosspeaks with deshielded 10m =>• OH group (n) • CH 9g has two crosspeaks with diastereotopic protons 4if 5 4 3 2 1 5 4 3 2 1 Q)2- 1H (ppm) Task 1: J-con necti vity of C10H20O Task 1: J-connectivity of C10H20O • CH 10m connected with CH2 8ck • CH2 4if connected with CH2 7bj X 5 4 3 2 1 Q)2- 1H (ppm) Task 1: -/-connectivity of C10H20O Task 1: J-connectivity of C10H20O • CH2 8ck weakly coupled with CH2 7bj =>- closing ring • protons b and k coupled to CH 6h which is connected to methyl 3e • other expected crosspeaks in DQ-COSY c rowd ed / ove r I a p p ed, found topology confirmed in HMBC (3/4Jhc) a,, A H 2d b M E & 2 3 4 _i_ UnHc-Hm, DnHg-Hm" UnHk-Hm"" TTnTTm—TTn - 3 2 Q)2- 1H (ppm) Jan Novotný >0 0,0 4/5 Task 1: J-connectivity of C10H20O 3(H 35 A E Q. O. O co 40H 45H 5(H Jan Novotný _i_i_i_ j_i_i_ HMBC «nC6-Hin^\ W6-Hb UnC6-Hk—----v T UnC7-He^ UnC6-Hf h 30 UnC7-Hk h 35 UnC7-Hj UnC7-Hb UnC7-Hi h 40 UnC8-He UnC8-Hm UnC8-Hn UnC8-Hk „ft „. \ UnC8-Hb UnC8-Hh \ ^u*iv*o—au h 45 UnC9-Hn UnC9-Hm \ \ UnC9:H1 UnC9-Hd "n^"f \ UnC9-Hi \ Un""Ha UneQ-HgS1 □ S - h 50 Task 1: J-connectivity of C10H20O • CH2 8ck weakly coupled with CH2 7bj =^> closing ring • protons b and k coupled to CH 6h which is connected to methyl 3e • other expected crosspeaks in DQ-COSY c rowd ed / ove r I a p p ed, found topology confirmed in HMBC (3/4JHc) H MENTHOL 3CH3 Jan Novotný 30- 35- E Q. Ü CO 40 45 50 UnC6-Hh UnC6-He HMBC UnC6-Hi UnC6-Hk UnC7-Hk UnC6-Hb UnC7-He^ UnC6-Hf UnC7-Hb - 35 UnC7-Hi -40 UnC8-Hn UnC8-He UnC8-Hm UnC8_Hk \ kUnC8-Hh -Hn \ xUnC8-Hh \ * \ \ \\V tTnP9-Hl UnC8-Hc UnC9-Hn UnC9-Hm uncyjii UnC9_Hd 1 \ \ UnC9-Hi\Un^ \ \ \ - \ ^/ UnC9-Hg UnC9-Ha 30 45 50 4 2 C02-1H (ppm) 4/5 Task 1: Stereochemistry of menthol C10H20O • lo2: homonuclear/heteronuclear couplings • large couplings preserved in ID slices of HSQC: axial H - 2 visible interactions (geminal and vicinal) x equatorial H - only geminal • ID TOCSY: selective decoupling =^ simplification of complex multiplets • DQF-COSY: analysis of phase sensitive spectrum • lo3: NOE contacts (axial strong) hx hb 3JX.A = small 3JX.B = large C hb 3JHC= 6-8 Hz 2JHr = 5-7 HZ hb 3JX.A = small 3JX.B = small C = 1-3 HZ R 2JHr = 0-2 Hz Jan Novotný 5/5 Task 1: Stereochemistry of menthol C10H20O • lo2: homonuclear/heteronuclear couplings • large couplings preserved in ID slices of HSQC: axial H - 2 visible interactions (geminal and vicinal) x equatorial H - only geminal • ID TOCSY: selective decoupling =^ simplification of complex multiplets • DQF-COSY: analysis of phase sensitive spectrum • lo3: NOE contacts (axial strong) Jan Novotný >0 0,0 5/5