基于固体核磁共振多次交叉极化的定量检测优化技术

doi:10.11938/cjmr20223039

摘要/Abstract

摘要：

多次交叉极化（multiCP）是一种耗时短、可行性高的固体核磁共振定量方法，近年来受到了广泛的关注．通过研究发现，multiCP实验参数的设置依赖于样品及基团的属性．对于样品属性差异较大的体系，其获取定量结果时的实验参数条件较苛刻．针对这一问题，本文结合LGCP（Lee-Goldburg cross polarization）技术，提出了一种multiCP的优化方案，命名为MLGCP-1．本文以L-丙氨酸、L-缬氨酸、L-丙氨酸/L-缬氨酸的混合物作为模型样品，并通过与multiCP进行对比，探讨MLGCP-1方法定量分析的可行性与优势．首先，通过对L-丙氨酸、L-缬氨酸样品基团比例的测量，发现MLGCP-1实现定量检测的参数—交叉极化接触时间（t_p）范围较multiCP更宽，可由1.0~1.3 ms增至0.8~2.0 ms．此外，通过对L-缬氨酸、L-丙氨酸/L-缬氨酸中特定基团间积分比值的分析发现，MLGCP-1与multiCP相似，同样受到¹³C-¹H交叉弛豫时间（T_CH）差异度的影响．即T_CH差异度越大，可定量的t_p参数范围越小．但与multiCP相比，MLGCP-1对t_p的宽容度更高．即对于相同的样品体系，MLGCP-1可定量的t_p范围更宽．总之，与multiCP相比，MLGCP-1可实现定量表征的实验参数范围更宽，更适用于表征属性差异较大的样品体系．

关键词: 固体核磁共振（SSNMR）, 交叉极化, 多次交叉极化, 定量方法, 氨基酸

Abstract:

Recently, multiple-cross polarization (multiCP) has attracted much interest owing to its favorable performance as a solid-state nuclear magnetic resonance quantitative method. Relating investigations revealed that the setup of multiCP parameters relies on the properties of the samples. Diverse types of samples require different parameters. To improve the tolerance to sample properties, an improved method named MLGCP-1 was proposed in this work, which employed Lee-Goldburg cross polarization technique. L-alanine, L-valine and their mixtures were chosen as model samples to evaluate the performance of MLGCP-1 method. multiCP experiments were also conducted for comparison. Based on the test of molecular group ratio, it was revealed that the range of contact time t_p of MLGCP-1 was larger than that of multiCP, which improved from 1.0~1.3 ms to 0.8~2.0 ms. Moreover, according to the study of L-valine and mixtures, it was revealed that the range of t_p was influenced by the difference of cross relaxation time T_CH. Large T_CHdifference limited t_prange for quantification. This manner was in accordance with multiCP. However, the t_prange of MLGCP-1 was markedly enlarged in comparison with multiCP, presenting higher tolerance to the sample properties.

Key words: solid-state nuclear magnetic resonance (SSNMR), cross polarization, multiCP, quantitative method, amino acid

中图分类号:

O482.53

董洪春,张志兰,王宁,唐丹丹,裘子慧,舒婕. 基于固体核磁共振多次交叉极化的定量检测优化技术[J]. 波谱学杂志, 2023, 40(2): 136-147.

DONG Hongchun,ZHANG Zhilan,WANG Ning,TANG Dandan,QIU Zihui,SHU Jie. The Improved Solid-state NMR Quantitative Method on the Bases of Multiple-cross Polarization Technique[J]. Chinese Journal of Magnetic Resonance, 2023, 40(2): 136-147.

图/表 10

图1

图2

表1

L-丙氨酸和L-缬氨酸分子中各基团在ramp-CP和LGCP条件下的TCH和 T 1 ρ H值

Sample	Carbons	T_1,H/ms	ramp-CP		LGCP
Sample	Carbons	T_1,H/ms	T_CH/ms	$T 1 ρ H$ /ms	T_CH/ms	$T 1 ρ H$ /ms
L-alanine	1	977.1	0.170	1.7	0.387	12.8
	2		0.042		0.174
	3		0.360		0.331
L-valine	4	718.5	0.174	2.5	0.414	24.3
	5		0.036		0.157
	6		0.033		0.120
	7		0.290		0.240
	8		0.154		0.374

表1

图3

图4

图5

图6

图7

图8

表2

参考文献 34

[1]	KOMOROSKI R A. High resolution NMR spectroscopy of synthetic polymers in bulk[M]. Florida: VCH. 1986.
[2]	MEHRING M. Principles of high resolution NMR in solids[M]. Berlin: Springer-Verlag. 1983.
[3]	DUER M J. Introduction to solid state NMR spectroscopy[M]. Oxford: Blackwell Science. 2004.
[4]	MAO J D, CAO X Y, OLK D C, et al. Advanced solid-state NMR spectroscopy of natural organic matter[J]. Prog Nucl Magn Reson Spectrosc, 2017, 100: 17-51. doi: 10.1016/j.pnmrs.2016.11.003
[5]	ZHANG L L, CHEN Q, HANSEN E W. Morphology and phase characteristics of high-density polyethylene probed by NMR spin diffusion and second moment analysis[J]. Macromol Chem Phys, 2005, 206(2): 246-257. doi: 10.1002/(ISSN)1521-3935
[6]	ZHANG L L, LIU Z, CHEN Q, et al. Quantitative determination of phase content in multiphase polymers by combining spin-diffusion and CP-MAS NMR[J]. Macromolecules, 2007, 40(15): 5411-5419. doi: 10.1021/ma0707786
[7]	LIU H W, ZHOU X Y, CHEN Q, et al. Accurate quantitative and maximum cross polarization via multiple ramped contacts[J]. Chem Phys Lett, 2017, 679: 233-236. doi: 10.1016/j.cplett.2017.05.009
[8]	SHU W F, ZHANG S M. Relaxation compensated and intensity recovered dynamics of cross polarization in the frame of reciprocity relation[J]. Chem Phys Lett, 2011, 511(4-6): 424-426. doi: 10.1016/j.cplett.2011.06.037
[9]	HOU G J, DING S W, ZHANG L M, et al. Breaking the T-1 constraint for quantitative measurement in magic angle spinning solid-state NMR Spectroscopy[J]. J Am Chem Soc, 2010, 132(16): 5538-5539. doi: 10.1021/ja909550f
[10]	HOU G J, DENG F, DING S W, et al. Quantitative cross-polarization NMR spectroscopy in uniformly C-13-labeled solids[J]. Chem Phys Lett, 2006, 421(4-6): 356-360. doi: 10.1016/j.cplett.2006.01.105
[11]	GU J L, ZHANG T T, ZHAO H P, et al. Time-saving and highly applicable quantitative method based on solid-state nuclear magnetic resonance techniques[J]. Chem J Chin Univ, 2018, 39(3): 463-469. doi: 10.1002/cjoc.v39.2
	顾佳丽, 张田田, 赵辉鹏, 等. 耗时短、通用性强的固体核磁共振交叉极化定量检测方法[J]. 高等学校化学学报, 2018, 39(3): 463-469. doi: 10.1002/cjoc.v39.2
[12]	SHU J, CHEN Q, ZHANG S M. Quantification of cross polarization with relaxation compensated reciprocity relation in NMR[J]. Chem Phys Lett, 2008, 462(1-3): 125-128. doi: 10.1016/j.cplett.2008.07.026
[13]	JOHNSON R L, SCHMIDT-ROHR K. Quantitative solid-state C-13 NMR with signal enhancement by multiple cross polarization[J]. J Magn Reson, 2014, 239: 44-49. doi: 10.1016/j.jmr.2013.11.009
[14]	DUAN P, SCHMIDT-ROHR K. Composite-pulse and partially dipolar dephased multiCP for improved quantitative solid-state C-13 NMR[J]. J Magn Reson, 2017, 285: 68-78. doi: 10.1016/j.jmr.2017.10.010
[15]	HOU G J, DENG F, YE C H, et al. Towards uniform enhancement in solid-state cross polarization magic angle spinning NMR: A scheme incorporating cross polarization with rotational resonance[J]. J Chem Phys, 2006, 124(23): 234512. doi: 10.1063/1.2206787
[16]	GERSTEIN B C, DYBOWSKI C R. Transient techniques in NMR of solids: An introduction to theory and practice[M]. San Diego: Academic Press. 1985.
[17]	ZHANG S M, WU X L, MEHRING M. Successive polarization under mismatched Hartmann-Hahn condition[J]. Chem Phys Lett, 1990, 166(1): 92-94. doi: 10.1016/0009-2614(90)87056-W
[18]	RAYA J, PERRONE B, HIRSCHINGER J. Chemical shift powder spectra enhanced by multiple-contact cross-polarization under slow magic-angle spinning[J]. J Magn Reson, 2013, 227: 93-102. doi: 10.1016/j.jmr.2012.12.006 pmid: 23314257
[19]	RAYA J, HIRSCHINGER J. Sensitivity enhancement by multiple-contact cross-polarization under magic-angle spinning[J]. J Magn Reson, 2017, 281: 253-271. doi: S1090-7807(17)30167-2 pmid: 28662486
[20]	BERNARDINELLI O D, LIMA M A, REZENDE C A, et al. Quantitative C-13 MultiCP solid-state NMR as a tool for evaluation of cellulose crystallinity index measured directly inside sugarcane biomass[J]. Biotechnol Biofuels, 2015, 8(110): 1-11. doi: 10.1186/s13068-014-0179-6
[21]	SAIDI F, TAULELLE F, MARTINEAU C. Quantitative C-13 solid-state NMR spectra by multiple-contact cross-polarization for drug delivery: From active principles to excipients and drug carriers[J]. J Pharm Sci, 2016, 105(8): 2397-2401. doi: 10.1016/j.xphs.2016.05.025
[22]	DUARTE R M B O, DUAN P, MAO J D, et al. Exploring water-soluble organic aerosols structures in urban atmosphere using advanced solid-state C-13 NMR spectroscopy[J]. Atmos Environ, 2020, 230: 117503. doi: 10.1016/j.atmosenv.2020.117503
[23]	CHU W Y, CAO X Y, SCHMIDT-ROHR K, et al. Investigation into the effect of heteroatom content on kerogen structure using advanced C-13 solid-state nuclear magnetic resonance spectroscopy[J]. Energ Fuels, 2019, 33(2): 645-653. doi: 10.1021/acs.energyfuels.8b01909
[24]	ZHANG Y L, LI L J, YAO S H, et al. Distinct changes in composition of soil organic matter with length of cropping time in subsoils of a Phaeozem and Chernozem[J]. Eur J Soil Sci, 2018, 69(5): 868-878. doi: 10.1111/ejss.2018.69.issue-5
[25]	WANG X Y, HAN X X, YOU Y L, et al. Molecular characterization of Dachengzi oil shale kerogen by multidimensional solid-state nuclear magnetic resonance spectroscopy[J]. Fuel, 2021, 303(6): 121215. doi: 10.1016/j.fuel.2021.121215
[26]	ZHANG Z L, WANG N, TANG D D, et al. Experimental set-up and application research of solid-state nuclear magnetic resonance Multiple-CP technique[J]. Chem J Chin Univ, 2021, 42(3): 784-793. doi: 10.7503/cjcu20200698
	张志兰, 王宁, 唐丹丹, 等. 固体核磁共振Multiple-CP定量技术的参数优化与应用研究[J]. 高等学校化学学报, 2021, 42(3): 784-793. doi: 10.7503/cjcu20200698
[27]	GOLDBURG W I, LEE M. Nuclear magnetic resonance line narrowing by a rotating RF field[J]. Phys Rev Lett, 1963, 11(6): 255-258. doi: 10.1103/PhysRevLett.11.255
[28]	LEE M, GOLDBURG W I. Nuclear-magnetic-resonance line narrowing by a Rotating RF field[J]. Phys Rev, 1965, 140(4a): 1261-1271.
[29]	PEERSEN O B, SMITH S O. Rotational resonance NMR of biological membranes[J]. Concepts Magn Reson, 1993, 5(4): 303-317. doi: 10.1002/(ISSN)1099-0534
[30]	BENNETT A E, RIENSTRA C M, AUGER M, et al. Heteronuclear decoupling in rotating solids[J]. J Chem Phys, 1995, 103(16): 6951-6958. doi: 10.1063/1.470372
[31]	MORCOMBE C R, ZILM K W. Chemical shift referencing in MAS solid state NMR[J]. J Magn Reson, 2003, 162(2): 479-486. pmid: 12810033
[32]	LADIZHANSKY V, VEGA S. Polarization transfer dynamics in Lee-Goldburg cross polarization nuclear magnetic resonance experiments on rotating solids[J]. J Chem Phys, 2000, 112(16): 7158-7168. doi: 10.1063/1.481281
[33]	FU R, HU J, CROSS T. Towards quantitative measurements in solid-state CPMAS NMR: A Lee-Goldburg frequency modulated cross-polarization scheme[J]. J Magn Reson, 2004, 168(1): 8-17. pmid: 15082244
[34]	JOHNSON R, SCHMIDT K. Quantitative solid-state ¹³C NMR with signal enhancement by multiple cross polarization[J]. J Magn Reson, 2014, 239: 44-49. doi: 10.1016/j.jmr.2013.11.009

样品组分	t_p/ms	MLGCP-1百分误差/%			multiCP百分误差/%
样品组分	t_p/ms	2/5	2/6	3/6	2/5	2/6	3/6
1:3	0.5	1.29	0.94	14.4	4.54	4.40	18.2
	1.0	1.80	4.57	4.53	0.85	8.35	10.2
	1.5	4.91	3.88	4.05	0.22	12.5	28.5
	2.0	3.63	1.27	10.1	0.58	18.7	51.0
1:1	0.5	1.24	1.84	9.67	0.49	1.31	20.1
	1.0	1.33	1.40	3.16	1.14	2.14	3.96
	1.5	1.09	1.13	3.54	0.83	2.63	12.7
	2.0	0.77	0.85	4.40	3.71	0.70	23.1
3:1	0.5	0.72	2.09	6.99	2.31	0.46	11.9
	1.0	2.14	1.76	2.34	0.68	4.99	4.09
	1.5	0.06	1.05	1.10	0.72	7.29	9.63
	2.0	0.92	1.48	0.74	0.74	9.98	14.2