环球仪器公司C18 SPE 固相萃取柱的应用

我们实验室在摸索了10种磺酰脲除草剂的前处理方法后，确定了采用固相萃取技术（SPE）小柱对土壤基质进行净化，方法建立了以后，与环球仪器公司合作使用其公司制作的C18 SPE 固相萃取小柱作为净化过程中使用的柱子。

其应用主要集中在单个磺酰脲除草剂的不同土壤基质中的前处理过程中，前处理过程如下： 提取:取10克土壤（过20目筛后风干）于50ml具塞的离心管中，添加不同浓度的1ml磺酰脲类除草剂的混标于离心管中,放置10min。加提取液PB7.8:乙腈=8:2 体积为10ml,涡旋2min,超声5min,离心4000~8000r/min 10min。反复提取3次,合并上清液.用磷酸480µl左右调节pH值在2.5左右,(在空白样品中测得磷酸体积,以免提取的样品损失)。

活化:用5ml乙腈浸泡C18 6ml小柱30min,用调节到2.5的提取液5ml预淋洗。

上样:在小柱液面未干之前上样,控制流速为0.5ml/min,上样1个小时,抽真空10min,除去多余水分。

洗脱:用乙腈:PB7.8=9:1的溶剂5ml洗脱,N2吹,定容到5ml,低浓度的用3ml洗脱,N2吹至1ml以下,并定容至1ml。加大浓缩力度，可使浓度的检出限更低.

方法建立初始采用的10种磺酰脲除草剂的结构式和相关性质

产品名称 原药含量%

结构式

Mr

分子式 性质

氯磺隆 92% 357.8 C12H12ClN5O4S

381.36 C14H15N5O6S

甲磺隆 98.8%

395.4

苯磺隆 95%

C15H17N5O6S

甲嘧磺隆95% 364.4 C14H14N4O5S

吡嘧磺隆99% 414.4 C14H18N6O7S

胺苯磺隆 98% 410.4 C15H18N6O6S

387.4

噻磺隆 95%

C11H11N5O6S2

氯嘧磺隆96% 414.82 C15H15ClN4O6S

烟嘧磺隆95.6% 410 C15H18N6O6S

苄嘧磺隆97% 410.40 C16H18N4O7S

做了3个水平的添加.实验结果如下表: Recovery 1 2 3 4 5 6

10ppm 1.04 1.11 1.12 1.14 0.92 1.10

1ppm 0.97 0.87 0.88 0.94 0.86 0.93

0.1ppm 0.95 0.88 1.04 0.93 0.91 1.03

7 8 9 10

0.37 0.85 0.94 1.16

0.17 0.83 0.86 0.86

线性关系与相关系数

0.35 0.90 1.01 0.87

Table 7 Linear equations and correlation coefficients

Herbicide Linear range/(mg/ L) Linear r

equation 3

nicosulfuron 0.1~10.0 y=13.028x+5.0699 0.9989 thifensulfuron methyl 0.1~10.0 y=17.851x+3.8571 0.9986 metsulfuron methyl 0.1~10.0 y=17.851x+3.8571 0.9986 sulforneturon-methyl 0.1~10.0 y=10.051x+1.7622 0.9986 chlorsulfuron 0.1~10.0 y=5.6007x+1.9487 0.9985 ethametsulfuron 0.1~10.0 y=6.8508x+2.4757 0.9979 bensulfuron methyl 0.1~10.0 y=11.35x+2.4985 0.9979 tribenuron methyl 0.1~10.0 y=19.875x+4.5357 0.9986 pyrazosulfuron-methyl 0.1~10.0 y=12,401x+2.1205 0.9986 chlorimuron ethyl 0.1~10.0 y=25.577x-3.9065 0.9964

Y: peak area ; X: mass concentration , mg/ L.

选取1ppm添加回收的质谱图 标样 1ppm

Intens.

7x101.25

05060302.D: TIC +All MS

1.00

0.75

0.50

0.25

0.00

681012141618Time [min]

样品 1ppm

Intens.

7x101.25

05060302.D: TIC +All MS

1.00

0.75

0.50

0.25

0.00

9101112131415161718Time [min]

后来应用到了单个药品的实际样品的检测，主要做了三个药，甲嘧磺隆，烟嘧磺隆和噻吩磺隆。前处理过程相同，实验结果和谱图如下： 甲嘧磺隆的东北土壤测定 0.2ppm standard 甲嘧磺隆

Intens.x1070.750.500.25x105

2

0.750.500.250.00

12345Time [min]CX100812.D: EIC 364.4 +All MS, Smoothed (1.7,1, GA)CX100812.D: TIC +All MS

Intens.x1051.50

+MS, 3.7-4.3min #(274-318)

1.25

1.00

158.9

300.9

326.9

430.60.75

0.50

149.0

0.25

114.00.00

100150200250300350400450m/z132.9

177.0

204.9224.0

267.0253.0

279.0

348.8338.1

364.9

386.8413.0

446.6

有草稻田 1和2

Intens.x1070.80.60.40.2x104

CX100813.D: TIC +All MS

CX100813.D: EIC 364.4 +All MS420

1234567Time [min]

Intens.x1071.000.750.500.25x10443210

123456CX100814.D: TIC +All MS

CX100814.D: EIC 365; 387 +All MS7Time [min]

结果表明在高灵敏度质谱条件下有草稻田中未检出甲嘧磺隆没有被污染。 无草稻田 3和4

Intens.x1071.000.750.500.25x10443210

1234567Time [min]CX100815.D: EIC 365 +All MSCX100815.D: TIC +All MS

Intens.x1071.0

CX100816.D: TIC +All MS

0.5

x10443210

123456CX100816.D: EIC 365 +All MS7Time [min]

结果表明施药的稻田也未检出甲嘧磺隆，说明已经降解。

烟嘧磺隆的北京土壤测定 烟嘧磺隆 标样 1ppm TIC

Intens.x1061.4YMHL0301.D: TIC +All MS, Smoothed (3.6,1, GA)1.21.00.812345678Time [min]

EIC

Intens.x1043YMHL0301.D: EIC 432.8 +All MS, Smoothed (3.0,1, GA), Smoothed (4.2,1, GA)21012345678Time [min]

MS

Intens.x10454321050100124.9150181.7212.7226.8200.8200277.7+MS, 4.4min #439, Background Subtracted454.7410.8432.8255.7250308.7300338.9350367.9400450m/z

标样1ppm的响应值很高 样品作了两次重复

烟嘧磺隆 样品1 TIC Intens. 6 x10

86

YMHL1031.D: TIC +All MS

4

2

2

4

681012

Time [min]

EIC

Intens.x1044YMHL1031.D: EIC 432.8 +All MS, Smoothed (3.3,1, GA)321024681012Time [min]

MS

Intens.x1042.01.51.00.50.05069.184.8100114.7150200250300350400450m/z148.7212.6198.6277.6261.9342.9400.7428.7432.8454.6486.8289.8+MS, 4.4min #494, Background Subtracted

烟嘧磺隆 样品2 TIC

Intens.x1068YMHL1032.D: TIC +All MS64224681012Time [min]

EIC

Intens.x104YMHL1032.D: EIC 432.8 +All MS, Smoothed (3.8,1, GA)4321024681012Time [min]

MS

Intens.x1046289.9+MS, 4.3min #488, Background Subtracted4280.898.650100120.7146.6166.8185.8150200230.8261.7250300330.9368.9354.8388.9350400432.8454.8450492.9m/z0

能看到一个很小的峰，说明有很少量的检出，但无法准确定量检出太小。

噻吩磺隆试验的土壤是湖南土，湖南土属于红壤土有机质含量较高，采用C18 SPE 小柱净化也仍然有显著效果实验结果和图谱如下 土壤中1ppm添加回收实验

S1 1ppm添加

Intens.500040003000200010000200015001000500024681012141618Time [min]106051101.D: EIC 166.6 +MS2(387.7), Smoothed (3.9,1, GA), Smoothed (5.8,1, GA)06051101.D: TIC +All MSn

Intens.1.1500+MS2(387.7), 10.7-12.1min1000166.6500368.8050100140.6150200250300342.7350400450m/z S2

Intens.600040002000015001100006051102.D: TIC +All MSn06051102.D: EIC 166.6 +MS2(387.7), Smoothed (5.8,1, GA)50002Intens.1.10004681012141618Time [min]

+MS2(387.7), 10.9-11.9min166.6800600400368.8200140.6050100150200250300350400450m/z342.7386.6

S3

Intens.6000400020000125010007505002500

24681012141618Time [min]1

06051103.D: EIC 166.6 +MS2(387.7), Smoothed (5.8,1, GA)06051103.D: TIC +All MSn

Intens.800600400200050100146.6150166.6+MS2(387.7), 11.4min #355368.7300.8276.7250300391.9204.7200231.7350.8350400450m/z S4

Intens.600040002000

0125010007505002500

24681012141618Time [min]1

06051104.D: EIC 166.6 +MS2(387.7), Smoothed (5.8,1, GA)06051104.D: TIC +All MSn

Intens.1.1000800600400200050100140.6150200250270.7300340.7350400368.8166.6+MS2(387.7), 10.6-12.5min450m/z Standard 1ppm 20ul

Intens.300020001000

06004002000

24681012141618Time [min]06051107.D: EIC 166.7 +MS2(387.7), Smoothed (7.7,1, GA), Smoothed (11.6,1, GA)1

06051107.D: TIC +All MSn

Intens.1.400166.6300200100314.7050100150200250300350400340.7+MS2(387.7), 9.0-12.5min368.8450m/z recovery

S1 1.04 S2 1.02 S3 0.972 S4 0.950

0.1ppm 土壤添加回收实验 0.1ppm std

Intens.x1041.010.80.60.40.20.03456789101112Time [min]EIC 431.6 +All MS, Smoothed (2.3,1, GA)

Intens.x104+MS, 8.8min #938, Background Subtracted1.25431.61.000.75494.90.50166.6148.5107.5326.9184.6122.8100150204.4222.7246.5261.9271.8300.0387.7346.8412.90.2596.456.80.005080.6457.9475.0200250300350400450m/z

t-1

Intens.x10410.80.60.40.20.002468101214Time [min]

Intens.x104378.9+MS, 9.0min #567, Background Subtracted1.5300.8412.8166.61.0431.6292.8184.70.5148.4174.5159.6232.70.0100150200250300350400450m/z192.6210.5238.7255.7273.8264.8284.8313.9333.7372.4421.1350.9356.4364.7390.9441.8

t-2

Intens.x1041CX060105.D: EIC 431.6 +All MS, Smoothed (4.3,1, GA)0.80.60.40.20.06Intens.x1043.078910111213Time [min]

+MS, 9.1min #574, Background Subtracted2.5378.92.01.5310.91.0148.5166.568.880.7105.6118.6100150203.5185.4200217.4213.9250413.0431.6240.5269.3302.8318.6300350344.1457.5391.9370.8400450488.1477.70.5358.60.0m/z

t-3

Intens.1CX060106.D: EIC 431.6 +All MS, Smoothed (4.4,1, GA)800060004000200006Intens.x10478910111213Time [min]

+MS, 9.1min #558, Background Subtracted2.5378.92.01.51.0148.5413.0431.6166.6300.8250.6174.5196.6150200216.7238.7268.7250300350400450m/z276.8320.9334.8349.0369.0441.9420.7487.0476.00.5120.582.695.00.0100134.5

0.1ppm玉米添加回收实验 y-1

Intens.1CX060107.D: EIC 431.6 +All MS, Smoothed (4.9,1, GA)800060004000200005Intens.x104678910111213Time [min]

+MS, 9.1min #581, Background Subtracted1.51.0118.5431.6184.50.594.672.678.60.0100150200250300350400134.5148.5166.7204.6226.5250.7269.4284.5306.9331.0348.7387.2412.9448.8440.0471.2484.4450m/z

y-2

Intens.x1041CX060108.D: EIC 431.6 +All MS, Smoothed (4.1,1, GA)0.80.60.40.26Intens.x1041.25789101112Time [min]

+MS, 9.1min #580, Background Subtracted413.01.00184.50.75145.5161.5250.7269.5333.9293.8386.9431.5444.6471.8344.90.50129.5118.5208.6232.896.588.568.8281.8323.7365.7483.80.250.00100150200250300350400450m/z

y-3

Intens.8000160004000200004567891011Time [min]

Intens.x1041.50206.61.25+MS, 7.8min #422, Background Subtracted412.9431.61.00162.50.75250.7268.8308.8333.9226.6236.70.2597.70.00100150200250300350400450m/z423.0419.6451.9286.7373.8396.0444.9275.8317.8366.90.50130.5145.5170.5182.6

std

0.1ppm 174653t-1

125720

t-2 143501t-3 148775

0.719827 0.0691640.821635 0.851832 rsd

y-1

130239

y-2 145714y-3 125049

0.745701 0.061554 0.834306 0.715985

rsd