Antibiotics

Application Note
RIDASCREEN Chloramphenicol Art. No. R1505
RIDA Chloramphenicol column Art. No. R1508
Enzymes

Reagents
200 mM sodium phosphate buffer pH 7.5 (for 1 l, weigh 26.7 g Na2HPO4 x 2 H2O
and 7.8 g NaH2PO4 x 2 H2O, adjust to pH 7.5 with NaOH)
1 N NaOH
Methanol p.a.
demineralized water

Procedure
I. Sample preparation
Liquid enzymes
dilute liquid enzymes 1:2 (1+1) with 200 mM sodium phosphate buffer,
(e.g. 5 ml liquid enzymes + 5 ml 200 mM sodium phosphate buffer)
Solid enzymes
dissolve in demineralized water to a concentration of 10 mg/ml
dilute dissolved enzymes 1:2 (1+1) with 200 mM sodium phosphate buffer,
(e.g. 5 ml dissolved enzymes + 5 ml 200 mM sodium phosphate buffer)

13/11/11

II. Clean-up with RIDA Chloramphenicol column

Do not let the columns dry up!
1. place the column on a column rack or clamp stand
2. place a reservoir on top of the column e.g. a 10 ml tip or attach a suitable
adapter on top of the column and use a syringe with a volume of 10 ml
3. pass 10 ml of the diluted enzyme solution through the immunoaffinity column by
gravity (flow rate: approx. 1 drop/s)
4. rinse the column with 10 ml demineralized water
5. discard the flow-through solution
6. remove reservoir or syringe and rinse the column with 1 ml demineralized water
7. press some air through the column or apply light vacuum (approx. 10 s) to make
sure that all the residual fluids will be removed from the column
8. place a clean and closable vial directly below the column
9. elute slowly with 0.5 ml methanol p.a. (flow rate: approx. 1 drop/s) to ensure
complete elution of chloramphenicol
10. collect all eluate residues by pressing air thoroughly through the column or
absorb with vacuum for 30 s
11. add 4.5 ml of sample dilution buffer of RIDASCREEN Chloramphenicol
(Art. No. R1505) to the eluat (0.5 ml) and mix

R-Biopharm AG
An der neuen Bergstrae 17, 64297 Darmstadt, Germany
Phone: +49 (0) 61 51 - 81 02-0, Fax: +49 (0) 61 51 - 81 02-40
E-mail: info@r-biopharm.de, www.r-biopharm.com

Antibiotics

Application Note
RIDASCREEN Chloramphenicol Art. No. R1505
RIDA Chloramphenicol column Art. No. R1508
Enzymes

III. Detection of Chloramphenicol

as described in the RIDASCREEN Chloramphenicol manual

Remarks for storage of samples and sample extracts:

store methanolic eluates at -20 C (-4 F) and protected against light. Under these
conditions the eluates can be stored for 2 to 3 months
samples reconstituted in sample dilution buffer have to be analyzed on the same day

Enzymes

Dilution Factor

Limit of Detection

Recovery

25 ppt

79 %

solid

100

2.5 ppb

74 %

13/11/11

liquid

R-Biopharm AG
An der neuen Bergstrae 17, 64297 Darmstadt, Germany
Phone: +49 (0) 61 51 - 81 02-0, Fax: +49 (0) 61 51 - 81 02-40
E-mail: info@r-biopharm.de, www.r-biopharm.com

Antibiotics

Validation Report
RIDASCREEN Chloramphenicol Art. No. R1505
RIDA Chloramphenicol column Art. No. R1508
Enzymes

Sample preparation and test implementation was performed in accordance to the

application note chloramphenicol in enzymes 2013-11-11 and the RIDASCREEN
Chloramphenicol test kit instructions 12-09-20.
Formulation

Enzyme

Blank
Concentration
[ppt]

Spiked with 200 ppt

Concentration
[ppt]

Recovery

168

84 %

liquid

Fructozym P6-X2

liquid

Trenolin rouge DF

12*

173

87 %

liquid

Trenolin Super DF

10*

140

70 %

liquid

Trenolin Frio DF

17*

151

76 %

liquid

Trenolin Bukett DF

13*

159

80 %

Mean

13

158

79 %

Precision

13

Limit of Detection
(mean + 3x precision)

< 25

25

Coefficient of Variation
(precision / mean)

Formulation

solid

8%

Enzyme

Blank

Trenolin Opti DF

solid

LittoZym Extraction

solid

LittoZym Brillance

Mean
Precision
Limit of Detection
(mean + 3x precision)

Spiked with 20 ppb

Concentration
[ppt]

Concentration
[ppt]

Recovery

1454*

13775

69 %

< 2500

14149

71 %

1394*

16677

83 %

1424

14867

74 %

42

1579

2500

Coefficient of Variation
(precision / mean)

11 %

13/11/11

* Results below the measuring range. The result was determined by extrapolation.

R-Biopharm AG
An der neuen Bergstrae 17, 64297 Darmstadt, Germany
Phone: +49 (0) 61 51 - 81 02-0, Fax: +49 (0) 61 51 - 81 02-40
E-mail: info@r-biopharm.de, www.r-biopharm.com