Volatile Organic Compounds from Bacillus subtilis Czk1: Optimization of Extraction Conditions Based on HS-SPME-GC-MS

doi:10.11924/j.issn.1000-6850.casb2020-0331

Abstract

Abstract:

The aim is to establish a rapid method for determining volatile organic compounds of Bacillus subtilis Czk1. The volatile organic compounds of Bacillus Subtilis Czk1 were identified by using headspace solid phase microextraction gas chromatogram mass spectrometry with the total peak number and total peak area as indicators, and the optimal extraction conditions were determined by single factor and orthogonal test. In the end, the volatile organic compounds of Czk1 were analyzed under these conditions. The results showed that the optimum extraction condition was the SPME fibers of 50/30 μm DVB/CAR/PDMS, extraction temperature of 40°C, extraction time of 40 min, desorption time of 7 min, and temperature program condition of 4. 33 kinds of volatile organic compounds were gained under the optimized condition, and the main components included ketones (28.65%), phenols (13.51%), esters (9.17%), aldehydes (15.60%), alcohols (12.08%), hydrocarbons (4.49%), and other substances (10.58%). The extraction method can provide a reference for the analysis of volatile organic compounds in Bacillus subtilis Czk1.

Key words: Bacillus subtilis Czk1, volatile organic compounds, headspace solid phase microextraction (HS-SPME), gas chromatography-mass spectrometry (GC-MS), optimization of extraction conditions

CLC Number:

S432.1

Liang Yanqiong, Li Rui, Wu Weihuai, Tan Shibei, Xi Jingen, Zheng Jinlong, Lu Ying, He Chunping, Yi Kexian. Volatile Organic Compounds from Bacillus subtilis Czk1: Optimization of Extraction Conditions Based on HS-SPME-GC-MS[J]. Chinese Agricultural Science Bulletin, 2021, 37(11): 24-31.

Figures/Tables 11

References 32

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萃取头类型	纤维涂层	pH	老化温度	老化时间/h	分析对象
100 μm PDMS,Fusedslilica/SS	PDMS	2~10	250	0.5	非极性挥发对象
7 μmPDMS,Fusedslilica/SS	PDMS	2~11	320	1	非极性挥发对象
85 μm Polyacrylate,Fusedslilica/SS	polyacrylate	2~11	280	0.5	极性挥发对象
50/30 μmDVB/CAR/PDMS StableFlex/SS	DVB/CAR/PDMS	2~11	270	0.5	极性/半极性挥发对象
65 μm DVB /PDMS StableFlex/SS	PDMS/DVB+OC	2~11	250	0.5	极性/半极性挥发对象

萃取头类型	纤维涂层	pH	老化温度	老化时间/h	分析对象
100 μm PDMS,Fusedslilica/SS	PDMS	2~10	250	0.5	非极性挥发对象
7 μmPDMS,Fusedslilica/SS	PDMS	2~11	320	1	非极性挥发对象
85 μm Polyacrylate,Fusedslilica/SS	polyacrylate	2~11	280	0.5	极性挥发对象
50/30 μmDVB/CAR/PDMS StableFlex/SS	DVB/CAR/PDMS	2~11	270	0.5	极性/半极性挥发对象
65 μm DVB /PDMS StableFlex/SS	PDMS/DVB+OC	2~11	250	0.5	极性/半极性挥发对象

序号	升温程序
1	40℃,保持2 min,以4℃/min上升到150℃,保持1 min,再以10℃/min上升到250℃,保持4 min,280℃后运行2 min。
2	50℃,保持2 min,以4℃/min上升到150℃,保持1 min,再以10℃/min上升到250℃,保持4 min,280℃后运行2 min。
3	40℃,保持1 min,以5℃/min上升到90℃,保持1 min,再以8℃/min上升到200℃,保持1 min, 最后以30℃/min上升到250℃,保持1 min,280℃后运行2 min。
4	50℃,保持1 min,以5℃/min上升到90℃,保持1min,再以8℃/min上升到200℃,保持1 min, 最后以30℃/min上升到250℃,保持1 min,280℃后运行2 min。
5	50℃,保持2 min,以4℃/min上升到100℃,保持4 min,再以10℃/min上升到200℃,保持5 min, 最后以25℃/min上升到250℃,保持5 min,280℃后运行2 min。

序号	升温程序
1	40℃,保持2 min,以4℃/min上升到150℃,保持1 min,再以10℃/min上升到250℃,保持4 min,280℃后运行2 min。
2	50℃,保持2 min,以4℃/min上升到150℃,保持1 min,再以10℃/min上升到250℃,保持4 min,280℃后运行2 min。
3	40℃,保持1 min,以5℃/min上升到90℃,保持1 min,再以8℃/min上升到200℃,保持1 min, 最后以30℃/min上升到250℃,保持1 min,280℃后运行2 min。
4	50℃,保持1 min,以5℃/min上升到90℃,保持1min,再以8℃/min上升到200℃,保持1 min, 最后以30℃/min上升到250℃,保持1 min,280℃后运行2 min。
5	50℃,保持2 min,以4℃/min上升到100℃,保持4 min,再以10℃/min上升到200℃,保持5 min, 最后以25℃/min上升到250℃,保持5 min,280℃后运行2 min。

试验号	因数					总峰面积×10⁸	总峰个数/个
试验号	A萃取温度		B萃取时间	C解吸时间	D升温程序	总峰面积×10⁸	总峰个数/个
1	1	0		1	-1	3.69d	79b
2	1	1		-1	0	3.02d	82b
3	0	-1		1	0	7.77a	101a
4	0	1		0	-1	4.23cd	94a
5	0	0		-1	1	5.63bc	99a
6	-1	1		1	1	4.23cd	89a
7	-1	-1		-1	-1	3.56d	98a
8	1	-1		0	1	6.23b	96a
9	-1	0		0	0	4.56cd	90a