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Featured Application:LC-MS/MSによるRaptor C18 を用いたマルチクラス動物用医薬品分析

Raptor C18を用いたLC-MS/MSによる抗生物質の一斉分析

  • 高い分離効率と短い分析時間を実現
  • 様々な種類の残留抗生物質に対応できる一斉分析条件と各クラスの抗生物質に最適化した定量用分析条件:

- マクロライド、リンコサミドおよびストレプトグラミン系抗生物質 (図 1)

- アンフェニコール、テトラサイクリン系抗生物質 (図 2)

- キノロン系抗生物質 (図 3)

- ペニシリン、セファロスポリンおよびテトラサイクリン系抗生物質 (図 4)

- サルファ剤 (図 5)
  (イオノフォア抗生物質にはRaptor Biphenylを使用 [図 6])

食用動物への抗生物質の使用は、薬剤耐性菌を増加させる可能性があるため、公衆衛生上および安全上の懸念事項です。欧州連合(EU) の多くの国やカ ナダでは、治療目的以外での抗生物質の使用は禁止されており、米国では抗生物質の成長促進剤としての使用を減らす政策を実施しています。動物用 抗生物質の適切な使用を規制するため、米国食品医薬品局(FDA)は、様々な動物組織および食品(21 CFR Part 556)に対して最大残留濃度(MRL)を設けて います。この規制を満たすためには、異なるクラスの抗生物質に対して高感度かつ効率的で信頼性の高い分析法が必要となり、Raptor C18はこの分析法 の理想的な選択肢です。


Restek Raptor LC Columns

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Desacetyl cephapirin0.70150382.03111.92124.21
2.Sulfanilamide0.85200172.9893.0775.23
3.Amoxicillin0.92100366.24349.10208.07
4.Cephapirin1.3650424.17292.08124.14
5.Tildipirosin1.38200734.59561.45204.15
6.Desfuroyl ceftiofur cysteine disulfide1.40300549.16183.02126.00
7.Lincomycin1.5050407.32359.23389.28
8.Sulfadiazine1.5720251.18156.0492.08
9.Cefquinome1.73200529.19134.10125.12
10.Ampicillin1.7850350.19106.07160.06
11.Sulfathiazole1.7910256.16156.0392.08
12.Marbofloxacin1.8110363.2072.11320.10
13.Cefalexin1.82100348.10158.05174.05
14.Sulfapyridine1.8610250.13156.1092.08
15.Norfloxacin1.9620320.23276.20233.13
16.Ofloxacin1.9810362.21318.20261.15
17.Sulfamerazine2.0020265.08156.0392.08
18.Cefalonium2.01100459.16337.03123.10
19.Oxytetracycline2.0225461.27426.15443.32
20.Ciprofloxacin2.0420332.18288.22245.15
21.Cefacetrile2.09300362.07258.08178.01
22.Tulathromycin A2.11100806.65577.42420.31
23.Tetracycline2.2125445.28154.07427.32
24.Danofloxacin2.2320358.22340.16314.21
25.Enrofloxacin2.3210360.29316.22245.13
26.Orbifloxacin2.3510396.22352.17226.12
27.Thiamphenicol*2.38200354.16290.04184.98
28.Sulfamethazine2.3910279.23186.08124.08
29.Sulfamethizole2.5210271.17156.02108.02
30.Sulfamethoxypyridazine2.5610281.14156.03126.07
31.Sarafloxacin2.5910386.20342.20368.15
32.Difloxacin2.6510400.23356.17299.13
33.Cefazolin2.75100455.10323.06295.09
34.Spiramycin2.96200843.64540.36699.48
35.Pirlimycin3.0520411.32363.18327.21
36.Chlortetracycline3.0825479.27154.07371.06
37.Sulfachlorpyridazine3.1220285.05156.03108.09
38.Gamithromycin3.28100777.63619.52601.45
39.Sulfadoxine3.3910311.17156.03108.09
40.Sulfamethoxazole3.4620254.18155.98147.06
41.Cefoperazone3.52100646.26143.07148.02
42.Florfenicol*3.55200356.10336.02184.98
43.Sulfaethoxypyridazine3.5620295.17267.07156.03
44.Tilmicosin3.64100869.72696.50522.42
45.Sulfisoxazole3.7620268.14156.03113.10
46.Oxolinic acid3.945262.10244.06215.96
47.Chloramphenicol*4.00200321.16151.99257.04
48.Ceftiofur4.1150524.14241.08125.24
49.Erythromycin4.3125734.64576.40558.38
50.Sulfadimethoxine4.3610311.17156.09108.09
51.Sulfaquinoxaline4.4220301.18156.04108.02
52.Tylosin**4.67100916.62772.49598.36
53.Penicillin G5.07100335.18176.07160.07
54.Flumequine5.345262.15244.11202.03
55.Penicillin V5.56100351.10160.06114.07
56.Oxacillin5.99100402.15160.05114.06
57.Virginiamycin M16.1350526.43508.31355.10
58.Tylvalosin6.19501042.71814.46640.39
59.Cloxacillin6.39100436.15277.06160.05
60.Nafcillin6.6025415.19199.09171.06
61.Dicloxacillin6.96100470.11160.05311.02
*Acquired in negative ion mode
Multiclass Veterinary Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0506
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:35 °C
Sample
Diluent:Water
Conc.:5–300 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
4.500.46535
7.000.44555
7.010.49010
9.000.49010
DetectorMS/MS
Ion Mode:ESI+/ESI-
Mode:Scheduled MRM
InstrumentUHPLC
Notes1. Positive and negative polarity data were collected simultaneously from a single injection.
2. Amphenicol compounds (chloramphenicol, thiamphenicol, and florfenicol) were detected with negative polarity.
3. The MRM was scheduled at +/- 20 to 30 seconds for each analyte.
4. Multiclass antibiotics include penicillin, cephalosporin, tetracycline, sulfonamide, macrolide, lincosamide, streptogramin, amphenicol, and quinolone.
**The retention time for Tylosin is noted in the peak list; however, it was not included in the chromatogram.

図 1: Raptor C18を用いたマクロライド、リンコサミドおよびストレプトグラミン系抗生物質のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Tildipirosin1.01200734.59561.45204.15
2.Lincomycin1.1150407.32359.23389.28
3.Tulathromycin A1.19100806.65577.42420.31
4.Spiramycin1.41200843.64540.36699.48
5.Pirlymycin1.4920411.32363.18327.21
6.Gamithromycin1.50100777.63619.52601.45
7.Tilmicosin1.58100869.72696.50522.42
8.Erythromycin1.8025734.64576.40558.38
9.Tylosin1.87100916.62772.49598.36
10.Tylvalosin2.30501042.71814.46640.39
11.Virginiamycin M12.4550526.43508.31355.10
Macrolide, Lincosamide, and Streptogramin Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0502
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:40 °C
Sample
Diluent:Water
Conc.:20–200 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
3.000.42080
3.010.49010
5.000.49010
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC

図 2: Raptor C18を用いたアンフェニコール系、テトラサイクリン系抗生物質のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Oxytetracycline1.2825461.27426.15443.32
2.Tetracycline1.3425445.28154.07427.32
3.Thiamphenicol*1.48200354.16290.04184.98
4.Chlortetracycline1.5625479.27154.07371.06
5.Florfenicol*1.86200356.10336.02184.98
6.Chloramphenicol*1.95200321.16151.99257.04
*Acquired in negative ion mode.
Amphenicol and Tetracycline Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0504
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:40 °C
Sample
Diluent:Water
Conc.:25–200 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
3.000.42080
3.010.49010
5.000.49010
DetectorMS/MS
Ion Mode:ESI+/ESI-
Mode:MRM
InstrumentUHPLC
NotesTetracyclines and amphenicols were analyzed with ESI+ and ESI- mode, respectively.

図 3: Raptor C18を用いたキノロン系抗生物質のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Marbofloxacin1.2810363.2072.11320.10
2.Norfloxacin1.3220320.23276.20233.13
3.Ofloxacin1.3210362.21318.20261.15
4.Ciprofloxacin1.3520332.18288.22245.15
5.Danofloxacin1.4020358.22340.16314.21
6.Enrofloxacin1.4410360.29316.22245.13
7.Orbifloxacin1.4710396.22352.17226.12
8.Sarafloxacin1.5510386.20342.20368.15
9.Difloxacin1.5710400.23356.17299.13
10.Oxolinic acid2.235262.10244.06215.96
11.Flumequine2.785262.15244.11202.03
Quinolone Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0505
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:40 °C
Sample
Diluent:Water
Conc.:5–20 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
3.000.44060
3.010.49010
5.000.49010
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC

図 4: Raptor C18を用いたペニシリン、セファロスポリンおよびテトラサイクリン系抗生物質のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Desacetyl cephapirin0.68150382.03111.92124.21
2.Amoxicillin0.89100366.24349.10208.07
3.Cephapirin1.0550424.17292.08124.14
4.Desfuroyl ceftiofur cysteine disulfide1.06300549.16183.02126.00
5.Cefquinome1.16200529.19134.10125.12
6.Ampicillin1.1850350.19106.07160.06
7.Cefalexin1.19100348.10158.05174.05
8.Oxytetracycline1.2650461.27426.15443.32
9.Cefalonium1.29100459.16337.03123.10
10.Tetracycline1.3250445.28154.07427.32
11.Cefacetrile1.37300362.07258.08178.01
12.Cefazolin1.50100455.10323.06295.09
13.Chlortetracycline1.5450479.27154.07371.06
14.Cefoperazone1.69100646.26143.07148.02
15.Ceftiofur1.8550524.14241.08125.24
16.Penicillin G2.18100335.18176.07160.07
17.Penicillin V2.33100351.10160.06114.07
18.Oxacillin2.44100402.15160.05114.06
19.Cloxacillin2.56100436.15277.06160.05
20.Nafcillin2.6325415.19199.09171.06
21.Dicloxacillin2.76100470.11160.05311.02
Penicillin, Cephalosporin, and Tetracycline Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0500
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:40 °C
Sample
Diluent:Water
Conc.:25–300 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
3.000.42080
3.010.49010
5.000.49010
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC

図 5: Raptor C18 を用いたサルファ剤のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Sulfanilamide0.83200172.9893.0775.23
2.Sulfadiazine1.4520251.18156.0492.08
3.Sulfathiazole1.6010256.16156.0392.08
4.Sulfapyridine1.6710250.13156.1092.08
5.Sulfamerazine1.7920265.08156.0392.08
6.Sulfamethazine2.0710279.23186.08124.08
7.Sulfamethizole2.1110271.17156.02108.02
8.Sulfamethoxypyridazine2.1610281.14156.03126.07
9.Sulfachlorpyridazine2.5520285.05156.03108.09
10.Sulfadoxine2.7510311.17156.03108.09
11.Sulfamethoxazole2.7820254.18155.98147.06
12.Sulfaethoxypyridazine2.8420295.17267.07156.03
13.Sulfisoxazole2.9820268.14156.03113.10
14.Sulfadimethoxine3.3710311.17156.09108.09
15.Sulfaquinoxaline3.4020301.18156.04108.02
Sulfonamide Antibiotics on Raptor C18 by LC-MS/MS
LC_FS0501
ColumnRaptor C18 (cat.# 9304A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:40 °C
Sample
Diluent:Water
Conc.:10–200 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.1% Formic acid in water
B:0.1% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.49010
3.500.46040
3.510.49010
5.000.49010
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC

図 6:Raptor Biphenyl を用いたイオノフォア抗生物質のLC-MS/MSによる分析

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Lasalocid A1.92100613.42377.28595.40
2.Monensin2.12100693.50675.44461.30
3.Salinomycin2.19100773.57431.24531.39
4.Maduramicin2.30100939.65877.58473.31
5.Narasin2.58100787.59431.27531.35
Ionophore Antibiotics on Raptor Biphenyl by LC-MS/MS
LC_FS0503
ColumnRaptor Biphenyl (cat.# 9309A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor Biphenyl EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9309A0252)
Temp.:40 °C
Sample
Diluent:Water:methanol (10:90)
Conc.:100 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:0.5% Formic acid in water
B:0.5% Formic acid in acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.53565
3.000.52575
3.010.53565
5.000.53565
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC