Analysis for water and feed additives

Claim 2

A PROCESS FOR THE ANALYSIS OF THE RESIDUAL GENTIAN VIOLET IN TISSUE WHICH COMPRISES: A. FINELY PARTICULATING A KNOWN AMOUNT OF TISSUE, B. ADMIXING A KNOWN AMOUNT OF FINELY PARTICULATED TISSUE AND A KNOWN AMOUNT OF WATER, C. ALLOWING THE ADMIXTURE TO STAND, D. EVAPORATING OR REMOVING THE WATER FROM THE ADMIXTURE, E. EVTRACTING ESSENTIALLY ALL OF THE FAT IN THE FINELY PARTICULATED TISSUE USING A FAT SOLVENT, F. EXTRACTING ESSENTIALLY ALL OF THE FAT REMAINING IN THE FINELY PARTICULATED TISSUE USING HEXANE, G. EVAPORATING OR REMOVING THE HEXANE FROM THE FINELY PARTICULATED TISSUE, H. ADMIXING A KNOWN AMOUNT OF ACIDIFIED ETHANOL WITH THE FINELY PARTICULATED TISSUE AND LETTING THE RESULTANT ADMIXTURE STAND, I. SEPARATING THE ACIDFIED ALCOHOL FROM THE RESULTANT ADMIXTURE AND J. DETERMINING THE AMOUNT OF GENTIAN VIOLET IN THE ACIDIFIED ALCOHOL BY SPECTROPHOTOMETRICAL COMPARISON WITH ACIDIFIED ALCOHOL STANDARDS CONTAINING SET AMOUNTS OF GENTIAN VIOLET, THE DETERMINED AMOUNT OF GENTIAN VIOLET BEING THE RESIDUE OF GENTIAN VIOLET IN THE KNOWN AMOUNT OF TISSUE.

Claim 3

A process as described in claim 1 wherein the fat solvent is petroleum ether.

Claim 4

A process as described in claim 1 wherein step (g) is achieved by centrifugation.

Claim 5

A process as described in claim 1 wherein step (e) is repeated twice, wherein step (f) is repeated twice, and wherein steps (h), (i) and (j) are repeated until the acidified alcohol from that sequence of steps (h), (i) and (j) contains no gentian violet, the residue of gentian violet in the known amount of tissue being the total of the determined amount of gentian violet in the acidified alcohol from each sequence of steps (h), (i) and (j).

Claim 6

A process for checking the accuracy of a process for the analysis of the residue gentain violet in tissue which comprises: a. preparing a known amount of finely particulated tissue known not to contain any gentian violet; b. admixing one or more batches of the particulated tissue with a known amount of an aqueous solution containing a known amount of gentian violet, each aqueous solution containing a different known amount of gentian violet to form spiked samples; c. extracting substantially all of the fat in each of the spiked samples using a fat solvent; d. extracting essentially all of the fat remaining in the spiked samples using hexane; e. evaporating or removing the hexane from each of the spiked samples; f. admixing a known amount of acidified ethanol with each of the spiked examples, and letting each of the resultant admixtures stand; g. separating the acidified alcohol from each of the resultant admixtures; and h. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet, the determined amount of gentian violet for each sample giving the accuracy of the procedure for the analysis of residual gentian violet in tissue by direct comparison with the known amount of gentian violet placed in each sample in step (b).

Claim 7

A process for checking the accuracy of a process for the analysis of the residue gentain violet in tissue which comprises: a. preparing a known amount of finely particulated tissue known not to contain any gentian violet; b. admixing one or more batches of the particulated tissue with a known amount of an aqueous solution containing a known amount of gentian violet, each aqueous solution containing a different known amount of gentian violet to form spiked samples; c. extracting substantially all of the fat in each of the spiked samples using a fat solvent; d. extracting essentially all of the fat remaining in the spiked samples using hexane; e. evaporating or removing the hexane from each of the spiked samples; f. admixing a known amount of acidified ethanol with each of the spiked examples, and letting each of the resultant admixtures stand; g. separating the acidified alcohol from each of the resultant admixtures; and h. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet, the determined amount of gentian violet for each sample giving the accuracy of the procedure for the analysis of residual gentian violet in tissue by direct comparison with the known amount of gentian violet placed in each sample in step (b).

Claim 8

A process as described in claim 5 wherein a blank sample of egg which contains no gentian violet is treated the same as each of the spiked samples in steps (a) to (f), and wherein the acidified ethanol resulting from the treatment of each of the spiked samples is compared with the acidified ethanol batch resulting from the treatment of the blank sample, this being a check on the acidified ethanol extraction steps.

Claim 9

A process as described in claim 5 wherein a blank sample of finely particulated tissue which contains no gentian violet is treated the same as each of the spiked samples in steps (b) to (g), and wherein the acidified ethanol resulting from the treatment of each of the spiked samples is compared with the acidified ethanol batch resulting from the treatment of the blank sample, this being a check on the acidified ethanol extraction steps.

Claim 10

A process as described in claim 7 wherein step (c) is repeated twice, wherein step (d) is repeated twice, and wherein steps (f), (g) and (h) are repeated until the acidified alcohol from the sequence of steps (f), (g) and (h) contains no gentian violet, the residue of gentian violet in the known amount of tissue being the total of the determined amount of gentian violet in the acidified alcohol from each sequence of steps (f), (g), and (h).

Claim 11

A process as described in claim 7 wherein step (b) is repeated twice, wherein step (c) is repeated twice, and wherein steps (e), (f), and (g) are repeated until the acidified alcohol from the sequence of steps (e), (f) and (g) contains no gentian violet, the residue of gentian violet in the known amount of egg being the total of the determined amount of gentian violet in the acidified alcohol from each sequence of steps (e), (f) and (g).

Claim 12

A process for the analysis of the gentian violet in feed or manure which comprises: a. extracting substantially all of the fat in the feed or manure using a fat solvent; b. drying said fat-extracted feed or manure; c. admixing a known amount of acidified ethanol with the fat-extracted feed or manure, and letting the resultant admixture stand; d. separating the acidified alcohol from the resultant admixture; e. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing set amounts of gentian violet; and f. repeating steps (c) to (e) until no gentian violet is found to be present in the acidified alcohol, the total of the gentian violet found from the determination steps being the total amount of gentian violet present in said feed or said manure.

Claim 13

A process for the analysis of the gentian violet in feed or manure which comprises: a. extracting substantially all of the fat in the feed or manure using a fat solvent; b. drying said fat-extracted feed or manure; c. admixing a known amount of acidified ethanol with the fat-extracted feed or manure, and letting the resultant admixture stand; d. separating the acidified alcohol from the resultant admixture; e. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing set amounts of gentian violet; and f. repeating steps (c) to (e) until no gentian violet is found to be present in the acidified alcohol, the total of the gentian violet found from the determination steps being the total amount of gentian violet present in said feed or said manure.

Claim 14

A process as described in claim 10 wherein the amount of gentian violet present in the manure is compared with the amount of gentian violet present in the feed before it is fed to said animal or said poultry, the difference being a measure of the gentian violet retained by said animal or said poultry.

Claim 15

A process as described in claim 10 wherein said feed is a poultry feed and said manure is poultry manure and wherein said fat extraction step involves first extracting part of said fat from said poultry manure of said poultry feed at least once with petroleum ether and then extracting substantially all of the remaining portion of said fat from said poultry manure or said poultry feed at least once with hexane.

Claim 16

A process as described in claim 12 wherein step (d) is achieved by centrifugation.

Claim 17

A process for checking the accuracy of a process for the analysis of the gentian violet in feed or manure which comprises; a. adding a known amount of various aqueous solutions containing a known amount of gentian violet to known amounts of feed or manure known not to contain any gentian violet to form spiked samples; b. extracting substantially all of the fat in each of the spiked samples of feed or manure using a fat solvent; c. admixing a known amount of acidified ethanol with each of the spiked examples, letting the resultant admixture stand; d. separating the acidified alcohol from each of the resultant admixtures; e. determining the amount of gentian violet in each of the acidified alcohols by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet; and f. repeating steps (c) to (e) until no gentian violet is found to be present in the acidified alcohol, the total of gentian violet for each sample giving the accuracy of the procedure for the analysis of residual gentian violet in said manure or said feed by direct comparison with the known amount of gentian violet placed in each sample in step (a).

Claim 18

A process for checking the accuracy of a process for the analysis of the gentian violet in feed or manure which comprises; a. adding a known amount of various aqueous solutions containing a known amount of gentian violet to known amounts of feed or manure known not to contain any gentian violet to form spiked samples; b. extracting substantially all of the fat in each of the spiked samples of feed or manure using a fat solvent; c. admixing a known amount of acidified ethanol with each of the spiked examples, letting the resultant admixture stand; d. separating the acidified alcohol from each of the resultant admixtures; e. determining the amount of gentian violet in each of the acidified alcohols by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet; and f. repeating steps (c) to (e) until no gentian violet is found to be present in the acidified alcohol, the total of gentian violet for each sample giving the accuracy of the procedure for the analysis of residual gentian violet in said manure or said feed by direct comparison with the known amount of gentian violet placed in each sample in step (a).

Claim 19

A process as described in claim 14 wherein one of said spiked samples contains no gentian violet so as to be a blank sample, wherein said acidified ethanol batches from the spiked samples are compared with the acidified ethanol batch from the blank sample, this being a check on the acidified ethanol extraction steps.

Claim 20

A process as described in claim 14 wherein said feed is a poultry feed and said manure is poultry manure and wherein said fat extraction step involves first extracting part of said fat from said poultry manure or said poultry feed at least once with petroleum ether and then extracting substantially all of the remaining portion of said fat from said poultry manure or said poultry feed at least once with hexane.

Claim 21

A process as described in claim 16 wherein step (d) is achieved by centrifugation.

Claim 22

A process for the analysis of the residual gentian violet in egg which comprises; a. admixing a known amount of egg and a known amount of water; b. allowing the admixture to stand; c. evaporating or removing the water from the admixture; d. extracting essentially all of the fat in the egg using a fat solvent; e. extracting essentially all of the fat remaining in the egg using hexane; f. evaporating or removing the hexane from the egg; g. admixing a known amount of acidiFied ethanol with the egg and letting the resultant admixture stand; h. separating the acidified alcohol from the resultant admixture; and i. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing set amounts of gentian violet, the determined amount of gentian violet being the residue of gentian violet in the known amount of egg.

Claim 23

A process for the analysis of the residual gentian violet in egg which comprises; a. admixing a known amount of egg and a known amount of water; b. allowing the admixture to stand; c. evaporating or removing the water from the admixture; d. extracting essentially all of the fat in the egg using a fat solvent; e. extracting essentially all of the fat remaining in the egg using hexane; f. evaporating or removing the hexane from the egg; g. admixing a known amount of acidiFied ethanol with the egg and letting the resultant admixture stand; h. separating the acidified alcohol from the resultant admixture; and i. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing set amounts of gentian violet, the determined amount of gentian violet being the residue of gentian violet in the known amount of egg.

Claim 24

A process as described in claim 18 wherein the fat solvent is petroleum ether.

Claim 25

A process as described in claim 18 wherein step (g) is achieved by centrifugation.

Claim 26

A process as described in claim 18 wherein step (d) is repeated twice, wherein step (e) is repeated twice, and wherein steps (d), (h) and (i) are repeated until the acidified alcohol from that sequence of steps (d), (h) and (i) contains no gentian violet, the residue of gentian violet in the known amount of egg being the total of the determined amount of gentian violet in the acidified alcohol from each sequence of steps (d), (h) and (i).

Claim 27

A process for checking the accuracy of a process for the analysis of the residue of gentian violet in egg which comprises: a. admixing a known amount of egg known not to contain any gentain violet with a known amount of an aqueous solution containing a known amount of gentian violet, each aqueous solution containing a different known amount of gentian violet to form spiked samples; b. extracting substantially all of the fat in each of the spiked samples using a fat solvent; c. extracting essentially all of the fat remaining in the spiked samples using hexane; d. evaporating or removing the hexane from each of the spiked samples; e. admixing a known amount of acidified ethanol with each of the spiked samples; and letting each of the resultant admixtures stand; f. separating the acidified alcohol from each of the resultant admixtures; and g. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet, the determined amount of gentian violet for each sample giving the accuracy of the procedue for the analysis of residual gentian violet in egg by direct comparison with the known amount of gentian violet placed in each sample in step (a).

Claim 28

A process for checking the accuracy of a process for the analysis of the residue of gentian violet in egg which comprises: a. admixing a known amount of egg known not to contain any gentain violet with a known amount of an aqueous solution containing a known amount of gentian violet, each aqueous solution containing a different known amount of gentian violet to form spiked samples; b. extracting substantially all of the fat in each of the spiked samples using a fat solvent; c. extracting essentially all of the fat remaining in the spiked samples using hexane; d. evaporating or removing the hexane from each of the spiked samples; e. admixing a known amount of acidified ethanol with each of the spiked samples; and letting each of the resultant admixtures stand; f. separating the acidified alcohol from each of the resultant admixtures; and g. determining the amount of gentian violet in the acidified alcohol by spectrophotometrical comparison with acidified alcohol standards containing varied amounts of gentian violet, the determined amount of gentian violet for each sample giving the accuracy of the procedue for the analysis of residual gentian violet in egg by direct comparison with the known amount of gentian violet placed in each sample in step (a).