Restek
Home / Resource Hub / Technical Literature Library / 60を超えるマルチクラス抗生物質の分析

60を超えるマルチクラス抗生物質の分析

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

articleBanner
  • 高い分離効率と短い分析時間を実現
  • 残留抗生物質分析の汎用性とグローバルな適用性―定量用に各クラスのパネルの最適化が可能:
    • 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])

 

食用動物に対する抗生物質の使用は、薬剤耐性菌を発生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。EUおよびカナダの多くの国では、治療以外の目的での抗生物質の使用を禁止しており、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に、米国食品医薬品局(FDA)は、様々な動物組織および食品について最大残留限界(MRL)を設定しています(21CFR Part556)。この規則を満たすためには、さまざまな種類の抗生物質の高感度、効率的、信頼性の高い分析法が必要であり、Raptor C18LCカラムが理想的な選択肢です。

cgarm-img
LC_FS0506
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
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
Standard/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.

Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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

cgarm-img
LC_FS0502
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
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
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
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
Standard/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
NotesWant even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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

cgarm-img
LC_FS0504
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.
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
Standard/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.

Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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

cgarm-img
LC_FS0505
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
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
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
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
Standard/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
NotesWant even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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

cgarm-img
LC_FS0500
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
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
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
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
Standard/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
NotesWant even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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

cgarm-img
LC_FS0501
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
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
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
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
Standard/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

cgarm-img
LC_FS0503
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
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
Standard/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
NotesWant even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.
FSSS2276A-JP