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| Tetracyclines Extraction from Musculature by SPE |
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Method:
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Matrix
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Application-No.:
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SPE
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muscles
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302030
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| Substances: |
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tetracycline; oxytetracycline; chlortetracycline |
| Column: |
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CHROMABOND® Tetracycline, 6 ml, 500 mg |
| Phase: |
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CHROMABOND® Tetracycline |
| MN catalog number: |
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730315 |
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| Sample pretreatment: |
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Weigh 10 g of a cut up sample in a centrifuge glass and add 93 g succinate
buffer 5.0 g succinic acid anhydride in 1 l dist. water, adjust with 1 M NaOH
to pH 4). Mix intensively (ultra turrax, 2 min), homogenise in an ultrasonic
bath (3 min) and centrifuge (5000 g, 15 min).
Aspirate 50 ml of this extract through a Cu-loaded chelating sepharose column.
Wash the column with 10 ml dist. water, 30 ml methanol and 2 x 10 ml dist.
water. Elute (4 ml/min) with 50 ml EDTA-succinate buffer (37.2 g Titriplex III
* H2O in 1 l succinate buffer)
Compounds investigated: Tetracycline, oxytetracycline, chlortetracycline
(100-500 mg/kg) |
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| Conditions: |
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Column conditioning:
1 column volume methanol, 1 column volume dist. water, then 1 column volume
EDTA-succinate buffer (see above)
CAUTION: DO NOT LET THE COLUMN RUN DRY !
Sample application:
Slowly force or aspirate the eluate of sample pretreatment through the column
Column washing:
2 ml dist. water (removal of Cu ions), 1 ml n-hexane.
Elution:
Elute with 7.5 ml methanol in a 25 ml tapered flask. Add 1 ml of an ethylene
glycol / methanol mixture (22 g ethylene glycol are filled up with methanol to
100 ml) and evaporate with a rotation evaporator (max. 40 °C) to dryness. Fill
up the residue with 0.1 M McIlvain-EDTA buffer (dissolve 52.5 g citric acid *
H2O, 44.5 g Na2HPO4 * H2O and 93 g Titriplex III in 2.5 l dist. water, adjust
with NaOH to pH 4) to 400 ml. |
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| Subsequent analysis: |
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HPLC column CC 250/4 NUCLEOSIL® 100-5 C18 HD , as described in MN Application No. 110710 |
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| Author(-s): |
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Lippold |
| Source: |
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Chemisches Landesuntersuchungsamt, Freiburg, Germany
(private communication) |
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| Keywords: |
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tetracyclines |
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Recovery rates:
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tetracycline, chlortetracycline ca. 50-70 %
oxytetracycline ca. 60-80 % |
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| www.mn-net.com |
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Concerning questions to literature applications please contact the cited author/s. Chemicals mentioned in applications are not manufactured or sold by MACHEREY-NAGEL. Applications mentioned in MACHEREY-NAGEL literature or databases are provided for informational purposes only. MACHEREY-NAGEL does not warrant that all applications have been tested in MACHEREY-NAGEL laboratories using MACHEREY-NAGEL products.
MACHEREY-NAGEL does not warrant the correctness of the applications. |
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