Determination of Lignin Monomer Crosslinking Structures in Wheat Stems by UPLC-MS/MS

doi:10.13560/j.cnki.biotech.bull.1985.2022-0002

Abstract

Abstract:

A method for the qualitative and quantitative determination of lignin monomer crosslinking structures in wheat stems by UPLC-MS/MS was established，providing a scientific basis for the development and utilization of lignin. The chromatographic separation was carried out on an ACQUITY UPLC^® BEH C18（2.1 mm×100 mm，1.7 μm），ultrapure water and acetonitrile were used as the mobile phase to conduct gradient elution，and MRM（multiple reaction monitoring）were employed for detection. The results showed that the crosslinking structures of 9 lignin monomers had good linearity in the linear correlation range（R²=0.998 9），the recovery was 91.67%-100%，and the relative standard deviations was 0.24%-2.10%. The method was applied to detect the crosslinking structure characteristics of 9 lignin monomers in wheat stems 14 d after jointing. The results revealed that the retention times of the crosslinking structures of 9 lignin monomers in the sample were the same as that of the standard one. G（8-8）G was the crosslinking structure of lignin monomers with the highest mass concentration，and the mass concentration range was 9.53-10.12 ng/g. In conclusion，the method is of strong specificity and high sensitivity. It is suitable for accurate qualitative and quantitative analysis of the crosslinking structures of nine lignin monomers in the wheat stems.

Key words: UPLC-MS/MS, wheat, stem, crosslinking structure of lignin monomer, recovery

SUN Shu-fang, LUO Yong-li, LI Chun-hui, JIN Min, XU Qian. Determination of Lignin Monomer Crosslinking Structures in Wheat Stems by UPLC-MS/MS[J]. Biotechnology Bulletin, 2022, 38(10): 66-72.

Figures/Tables 7

References 29

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时间Time/min	流速Flow/（mL·min^-1）	流动相A Mobile phase A/%	流动相B Mobile phase B/%	梯度变化Gradient
Initial	0.4	95.0	5.0	线性Linear
0.50	0.4	95.0	5.0	线性Linear
3.00	0.4	75.0	25.0	线性Linear
3.50	0.4	10.0	90.0	线性Linear
4.00	0.4	10.0	90.0	线性Linear
4.10	0.4	95.0	5.0	线性Linear
6.00	0.4	95.0	5.0	线性Linear

时间Time/min	流速Flow/（mL·min^-1）	流动相A Mobile phase A/%	流动相B Mobile phase B/%	梯度变化Gradient
Initial	0.4	95.0	5.0	线性Linear
0.50	0.4	95.0	5.0	线性Linear
3.00	0.4	75.0	25.0	线性Linear
3.50	0.4	10.0	90.0	线性Linear
4.00	0.4	10.0	90.0	线性Linear
4.10	0.4	95.0	5.0	线性Linear
6.00	0.4	95.0	5.0	线性Linear

待测物 Analyte	ESI模式 ESI model	保留时间 Retention time/min	母离子 Parent ion/(m/z)	子离子 Daughter ion/(m/z)	锥孔电压 Cone/V	碰撞能量 Collision/V
S（8-O-4）S（8-8）S	-	4.08	643.00	225^*/417.31	35	20/16
S（8-O-4）G（8-O-4）S	-	4.00	631.00	209.17^*/421.08	40	25/12
G（8-O-4）S（8-5）G	-	4.04	583.00	195.02^*/369.21	33	18/25
S（8-O-4）S（8-5）G	-	4.04	613.00	225.15^*/369.16	35	20/19
G（8-O-4）S（8-8）S	-	4.08	613.52	195.12^*/417.03	45	25/22
G（8-O-4）G（8-5）G	-	4.01	553.00	194.88^*/339.00	50	23/19
S（8-O-4）S	-	4.01	435.00	198.08^*/225.00	44	24/18
G（8-8）G	-	4.03	357.00	357.00	35	25/23
G（8-5）H	-	4.05	327.00	309.00^*/190.73	40	26/24

待测物 Analyte	ESI模式 ESI model	保留时间 Retention time/min	母离子 Parent ion/(m/z)	子离子 Daughter ion/(m/z)	锥孔电压 Cone/V	碰撞能量 Collision/V
S（8-O-4）S（8-8）S	-	4.08	643.00	225^*/417.31	35	20/16
S（8-O-4）G（8-O-4）S	-	4.00	631.00	209.17^*/421.08	40	25/12
G（8-O-4）S（8-5）G	-	4.04	583.00	195.02^*/369.21	33	18/25
S（8-O-4）S（8-5）G	-	4.04	613.00	225.15^*/369.16	35	20/19
G（8-O-4）S（8-8）S	-	4.08	613.52	195.12^*/417.03	45	25/22
G（8-O-4）G（8-5）G	-	4.01	553.00	194.88^*/339.00	50	23/19
S（8-O-4）S	-	4.01	435.00	198.08^*/225.00	44	24/18
G（8-8）G	-	4.03	357.00	357.00	35	25/23
G（8-5）H	-	4.05	327.00	309.00^*/190.73	40	26/24

待测物Analyte	含量Content/（ng·g^-1）
S（8-O-4）S（8-8）S	0.03
S（8-O-4）G（8-O-4）S	0.04
G（8-O-4）S（8-5）G	0.55
S（8-O-4）S（8-5）G	0.05
G（8-O-4）S（8-8）S	0.29
G（8-O-4）G（8-5）G	0.03
S（8-O-4）G	0.05
G（8-8）G	9.79
G（8-5）H	0.15