Aela Agela Technologies

Advance search
Quick Order
See your previous items:
You have 0 product(s) in cart

Determination of Antibiotics in Water using Cleanert LDC (PEP-2)

Application Introduction

Antibiotics were called as anti-bacteria elements before. Actually they not only can kill bacteria, but also have excellent bacteriostatic and killing effects on moulds, mycoplasmas, chlamydiae,

Favorite

Introduction

Antibiotics were called as anti-bacteria elements before. Actually they not only can kill bacteria, but also have excellent bacteriostatic and killing effects on moulds, mycoplasmas, chlamydiae, spirochaete, rickettsiae and other pathogenic microorganisms, so anti-bacteria elements are renamed as antibiotics. Antibiotics may be the substances generated during the growth and reproduction of certain microorganisms. For the curative antibiotics, a part of them are directly extracted from the microorganisms, the other part of them are completely or partially artificially synthesized. Popularly speaking, antibiotics are the drugs for treating various non-virus infections. However, clinical application has shown many side effects of antibiotics. The heavy usage of antibiotics would bring relatively toxic side effects and direct damage to human bodies, especially for children’s hearing. The abuse of antibiotics would result in drug-resistance of bacteria and kill a lot of normal bacteria in human bodies, consequently leading to dysbacteriosis of normal flora in human bodies.

Therefore monitoring antibiotics in water through effective experimental technical means is of great importance to ensuring people’s health.

Based on the detection methods for antibiotics in water, this experiment establishes a LC/MS/MS method for confirming and quantitatively detecting various antibiotics in water meanwhile by optimizing the pretreatment method.

Experimental

Materials and Reagents

Chromatographic solvent: acetonitrile; experimental water: ultrapure water; Ammoniated methanol (V/V: 5%); Formic acid aqueous solution (V/V: 0.1%);

Cleanert® LDC (PEP-2): 1 g/200 mL;

Sample Preparation

Sample 500 mL of water; add 0.25 g of EDTA to it; adjust the pH value with hydrochloric acid to 4.0; store the mixed solution for further purification.

Activate the Cleanert LDC (PEP-2) special columns for large-volume water samples (1 g/200 mL) with 9 mL of methanol, 5 mL of formic acid aqueous solution (V/V: 0.1%) in turn; load the aforesaid solution to be purified to the columns at the flow rate of 90 mL/min, leach the columns with 5 mL of water, and keep pumping them for 15 min; elute the columns with 20 mL of ammoniated methanol (V/V: 5%), and collect the eluent; blow the eluent with nitrogen to dry at 45℃, and dissolve it with 1 mL of acetonitrile aqueous solution (V/V: 20%) to the specified volume; filter a proper amount of supernatant with a 0.22 µm syringe filter for the next LC/MS/MS detection.

Dilute the high-concentration pesticide-mixed standard solution with blank sample matrix solution to the matrix-mixed standard working solution (concentration: 0.01 µg/mL).

Instrumentation

HPLC Column: Unisol C18, 150mm × 2.1 mm, 5 μm, 100 Å

Mobile Phase: A: 0.1 % formic acid-Water; B: Acetonitrile

Injection volume: 10 µL

Column Temperature: 30 °C

Scanning mode: Positive ion scanning;

Ion source: ESI source;

ESI voltage: 5500 V;

Atomized gas pressure: 55 psi;

Curtain gas pressure: 20 psi;

Auxiliary gas pressure: 50 psi;

Ion source temperature: 650 °C;

Collecting mode: MRM.

Scanning mode: Negative ion scanning;

Ion source: ESI source;

ESI voltage: -3500 V;

Atomized gas pressure: 60 psi;

Curtain gas pressure: 15 psi;

Auxiliary gas pressure: 50 psi;

Ion source temperature: 600 °C;

Collecting mode: MRM.

Table 1 HPLC Gradient Elution Conditions

Time/min

Flow/(mL/min)

A/%

B/%

1.00

0.30

90

10

3.00

0.30

40

60

5.00

0.30

5

95

7.00

0.30

5

95

8.00

0.30

90

10

20.00

0.30

90

10

Table 2 HPLC Gradient Elution Condition on Negative Ion Mode

Time/min

Flow/(mL/min)

A/%

B/%

1.50

0.30

85

15

5.00

0.30

30

70

5.01

0.30

10

90

6.00

0.30

10

90

6.01

0.30

85

15

10.00

0.30

85

15

Table 3 MS Parameters of Antibiotics

Antibiotics

Q1

Q3

DP/V

CE/V

Erythrocin

734.6

158.1

81

49

576.8

86

49

Josamycin

828.3

174.3

121

45

109.3

126

45

Kitasamycin

772.4

215

116

41

174.1

121

41

Oxolinic   Acid

262

244

41

27

216

41

41

Flumequine

262

244

41

27

202

41

45

Nalidixic   Acid

233

215

40

19

187

40

35

Sulfapyridine

250

156

65

24

184

65

24

Sulfachloropyridazine

285

156

50

23

108

50

35

Sulfamethazine

265

156

60

25

172

60

25

Sulfamethizole

271

156

50

20

107

50

32

Sulfmethoxypyridazine

281

156

70

25

215

60

25

Sulfametoxydiazine

281

156

70

25

108

70

35

Florfenicol

356

336

-78

-23

185

-78

-23

Results and Discussion

As is shown in Table 1, the standard addition recoveries of in-water antibiotics detected by LC/MS/MS method based on SPE technology are between 60% and 120%, and the variable coefficients of the antibiotics are less than 20%, conforming to the detection requirements. As is shown in Figure 1, 2, 3, 4, 5, 6, 7 and 8, the detection results show the good purification effect of the Cleanert LDC (PEP-2) as well as the good peak shapes and stable retention time of the antibiotics detected with Unisol C18.

Table 1 Recoveries and Reproducibility of Antibiotics in Water Samples (0.02 µg/L Spiked Sample)

Antibiotics

Recoveries/%

CV/%

RT/min

Erythrocin

74.8

14.2

5.91

Josamycin

82.3

2.0

6.19

Kitasamycin

100.7

15.0

6.05

Oxolinic   Acid

92.6

14.0

7.11

Flumequine

104.2

6.3

6.58

Nalidixic   Acid

107.8

8.6

7.08

Sulfapyridine

94.9

10.7

5.65

Sulfachloropyridazine

75.5

0.3

6.31

Sulfamethazine

75.1

12.4

5.81

Sulfamethizole

95.5

14.1

5.98

Sulfmethoxypyridazine

80.6

7.9

6.01

Sulfametoxydiazine

68.3

4.6

6.19

Florfenicol

86.3

3.9

7.07

 

Figure 1 LC-MS/MS Chromatogram of 0.01 μg/mL Standard Solution Mixture in Positive Ion Mode

Figure 2 LC-MS/MS Chromatogram of 0.01 μg/mL Standard Solution Mixture in Negative Ion Mode

Figure 3 LC-MS/MS Chromatogram of Blank Sample in Positive Ion Mode

Figure 4 LC-MS/MS Chromatogram of Blank Sample in Negative Ion Mode

Figure 5 LC-MS/MS Chromatogram of 0.01 μg/mL Standard Solution Mixture in Water Matrix in Positive Ion Mode

Figure 6 LC-MS/MS Chromatogram of 0.01 μg/mL Standard Solution Mixture in Water Matrix in Negative Ion Mode

Figure 7 LC-MS/MS Chromatogram of 0.02 μg/L Spiked Water Sample in Positive Ion Mode

Figure 8 LC-MS/MS Chromatogram of 0.02 μg/L Spiked Water Sample in Negative Ion Mode

Conclusion

This experiment establishes a LC/MS/MS detection method for antibiotics in water and determines the contents of antibiotics in water on the basis of SPE technology. The sample added with standard solution (0.02 µg/L) is detected, and the resultant recoveries are between 60% and 120%, conforming to the requirement. The high stability of SPE method and good reproducibility of chromatographic columns show this LC/MS/MS detection method can be used for detecting the residues of antibiotics in water.

Ordering Information

Products

Specification

Cat.No

Cleanert® LDC (PEP-2)

1 g/200 mL

LPE000100-2

Unisol C18

2.1 × 150 mm, 5 μm, 100 Å

UO950502-2

SPE-M08 Positive   Pressure SPE Device

8 Positions

SPE-M08

Nitrogen blowing instrument

Maximal 15 sample

NV15-G

Guard cartridge holder

Suitable for 4.6×10 mm and 2.1×10 mm

SH-100

Direct-connected Guard Column

5 μm,100 Å, 2.1×10 mm

VA950102-0S

1.5 mL vials

Screw neck vials, 12 × 32 mm

AV1001-6

Caps and Septa

Screw neck cap, center hole; red silicone/white PTEE septa, slitted

AV2200-0

Syringe Filter (Nylon)

0.22 μm, diameter 13 mm, 200/pk

AS021320

Disposable Syringe

2 mL, 100/pk

LZSQ-2ML

 


Material Name Material Type Date
Modify product number success!
Total Price:

Close

Email To Sombody

X
Email
Subject
Content

Login

X
Email address
Password
Register Forgotten your password?