RAPID MEASUREMENT OF 1H 90 PULSE IN THE SOLID STATE NMR VIA CROSS POLARIZATION

Pellegrino Conte

pellegrino.conte@unina.it

Dipartimento di Scienze del Suolo, della Pianta e dellAmbiente Universit di Napoli Federico II Via Universit 100, 80055 Portici (Na), ITALY

Solid state NMR spectroscopy is widely applied in many fields:

1. Analysis of structures and molecular dynamics of technologically relevant polymers 2. Analyses of environmentally significative organic and inorganic materials 3. Control of the quality of industrial products such as pharmaceutical ones and polymer blends 4. Studies of reactions in the solid state

Solid state NMR spectroscopy in Soil Science

1. Analyses of the organic and inorganic soil components 2. Quantification of the different chemicals belonging to natural organic matter (humic substances) 3. Evaluation of the transformation of soil organic matter as affected by soil management, soil formation, soil transformation and so on

Classical CPMAS

13C

NMR sequence

Positive NMR Signals

1

Null NMR Signals

0 0

-1

Negatve NMR Signals

Classical 1H 90 pulse calibration on glycine

2.96 s

The 1H 90 pulse is that obtained by halving the 180 pulse measured by inversion recovery

Alternative method to measure 1H 90 pulse: P360 sequence

O
176 ppm 43 ppm

Glycine
OH
176

NH2
43

Classical CPMAS 13C spectrum with 1H 90 pulse of 2.96 s

P360 CPMAS 13C spectrum. 1H 360 pulse: 4 x 2.96 s= 11.84 s

300

200

100 Chemical Shift (ppm)

-100

Theoric pulse length

Pulse length affected by circuit impedance

Due to the circuit impedance, the inversion recovery experiment on glycine provided 1H 90 pulse that was too long. In fact, the P360 experiment revealed a residual NMR signal of both carbons of glycine.

4x90 pulse sequence to measure 1H 90 pulse

As suggested by the NMR producers for the liquid state NMR, distortion of pulses due to circuit impedances can be overcome by applying a pulse train made by small pulses

4x90 pulse sequence to measure 1H 90 pulse

Bruker language Classical CP sequence 4 x 90 sequence

O
176 ppm 43 ppm

Glycine
OH
176

NH2
43

Classical CPMAS 13C spectrum with 1H 90 pulse of 2.96 s P360 CPMAS 13C spectrum. 1H 360 pulse: 4 x 2.96 s= 11.84 s 4x90 CPMAS 13C spectrum. 1H 90 pulse: 2.80 s

300

200

100 Chemical Shift (ppm)

-100

O O

H2 O C
79 ppm

33 ppm

S O

C H2

H2 C 24 ppm CH3
14 ppm

SDS
33

n=9
24 14

79

Classical CPMAS 13C spectrum with 1H 90 pulse of 2.96 s P360 CPMAS 13C spectrum. 1H 360 pulse: 4 x 2.96 s= 11.84 s 4x90 CPMAS 13C spectrum. 1H 90 pulse: 2.80 s

300

200

100 Chemical Shift (ppm)

-100

OH

CMC
*

62 ppm 178 ppm

O

6
O

O H2C

74 C2-C6

4 5
HO

2 3
O

O O

n
104 ppm

104 178

62

A. CP spectrum of CMC obtained with a proton pulse of 2.80 s B. CMC spectrum acquired with the 4 x 90 pulse sequence and a 2.80 s proton pulse C. as in B. but with a proton pulse of 2.78 s;
300 200 100 Chemical Shift (ppm) 0 -100

CH3 H3C H3C CH3 CH3 CH3

HMB
A. CP spectrum of HMB with 1H pulse of 2.80 s B. 4 x 90 HMB spectrum with 1H pulse of 2.80 s C. 4 x 90 HMB spectrum with 1H pulse of 2.83 s.

Car
132

17

-CH3-

300

200

100 Chemical Shift (ppm)

-100

pepsine

71 74 92 172 107 61 54 40 33 25

A. CP spectrum of pepsine obtained with a proton pulse of 2.80 s B. pepsine spectrum acquired with the 4 x 90 pulse sequence and a 2.80 s proton pulse;

128

300

200

100 Chemical Shift (ppm)

-100

HA
33

129 102 173

52 72

A. CP spectrum of HA with 1H pulse of 2.80 s B. 4 x 90 HA spectrum with 1H pulse of 2.80 s C. 4 x 90 HA spectrum with 1H pulse of 2.78 s.

300

200

100 Chemical Shift (ppm)

-100

CONCLUSIONS 1. The standard calibration by inversion recovery provided a too long 1H 90 pulse 2. Only few attempts were needed to find the correct 1H 90 pulse by 4x90 pulse sequence 3. The 4x90 pulse sequence was successfully and rapidly applied on very complex organic systems such as a protein (pepsine) and a humic acid 4. The 4x90 pulse sequence can efficaciously replace the standard calibration with the inversion recovery routine