Simultaneous Determination of Jasmonic Acid and 12-Oxo- Phytodienoic Acid in Different Plant Materials by HPLC-MS/MS

doi:10.11983/CBB16156

Abstract

Abstract: Jasmonic acid is one of the important phytohormones responding to abiotic and biotic injury. Detecting the acid is difficult because of the low content in plant material. 12-oxo-phytodienoic acid is an important precursor in the biosynthesis of jasmonic acid. In this research, we established an HPLC-MS/MS method to determine the content of these two target chemicals simultaneously in 4 types of plant materials. An external standard curve was used to calculate the content in plant material. The regression coefficiency factor of the standard curve reached 0.991 9 to 0.999 5. Spiked recovery was from 91.91% to 101.56%. The method is sensitive and available; The limits of detection of two target chemicals were 3.218 and 34.129 pmol. This method was used to determinate the contents of these two target chemicals in 4 types of plant materials.

Key words: jasmonic acid, 12-oxo-phytodienoic acid, HPLC-MS/MS

Shuangyu Zhang, Qi Qi, Xiaohui Li, Ying Gai, Xiangning Jiang. Simultaneous Determination of Jasmonic Acid and 12-Oxo- Phytodienoic Acid in Different Plant Materials by HPLC-MS/MS[J]. Chinese Bulletin of Botany, 2017, 52(5): 631-636.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB16156

https://www.chinbullbotany.com/EN/Y2017/V52/I5/631

Figures/Tables 10

Table 1 HPLC gradient program

Time (min)	Formic acid (0.1%)	Methanol
0	90	10
3	90	10
37	0	100
42	0	100
45	90	10
50	90	10

Table 2 Mass spectrum parameters of two target chemicals

Compound	Scan mode	Precursor ion	Product ion (MS2)
Jasmonic acid	+	211	181
12-oxo-phytodienoic acid	-	292	273

Figure 1 The suitable collision energy (eV) for daughter ion of two target chemicals(A) Relative content of daughter ion 273 and parental ion 292 of 12-oxo-phytodienoic acid (OPDA); (B) Relative content of daughter ion 181 and parental ion 211 of jasmonic acid (JA)

Figure 2 Chromatogram of two standard chemicals(A) MS2 total ion chromatogram (TIC); (B) Selected ion chromatogram (SIC). The retention time of jasmonic acid (JA) was 26.11 min, and that of 12-oxo-phytodienoic acid (OPDA) was 38.82 min.

Figure 3 Chromatogram of two target chemicals in riceThe retention time of jasmonic acid (JA) was 26.11 min, and that of 12-oxo-phytodienoic acid (OPDA) was 38.82 min, which was the same with that of standard chemicals.

Figure 4 Mass spectrum and possible fragmentation of two standard chemicals(A) 12-oxo-phytodienoic acid (OPDA); (B) Jasmonic acid (JA)

Figure 5 Mass spectrum of two target chemicals in plant sample(A) 12-oxo-phytodienoic acid (OPDA); (B) Jasmonic acid (JA)

Table 3 Standard curve and limit of detection (LOD) of two target chemicals

Compound	Standard equation	R²	Limit of detection (pmol)	Linear range (pmol)
Jasmonic acid	Y=24802.4X	0.9995	3.218	3.218-824.232
12-oxo-phytodienoic acid	Y=1.70573e+006X	0.9919	34.129	34.129-341.297

Table 4 Spiked recoveries of two target chemicals (n=3)

Compound	Spiked recovery (%)
Compound	7.5 ng	15 ng	30 ng
Jasmonic acid	92.68±3.46	95.34±2.31	91.91±7.49
12-oxo-phytodienoic acid	92.33±9.75	96.10±5.76	101.56±7.41

Table 5 Contents of two target chemicals in 4 species of plant materials (n=3)

Plant sample	Content (ng∙mg^-1DW)
Plant sample	Jasmonic acid	12-oxo-phytodie- noic acid
Eucommia ulmoides (leaf)	2.184±0.318	0.542±0.030
Viola yedoensis (flower)	1.069±0.065	0.166±0.031
Oryza sativa (root)	2.269±0.061	0.041±0.005
Ziziphus jujuba (fruit)	2.999±0.245	0.018±0.002

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