クロルテトラサイクリンの化学的定量法-II : 新螢光定量法の簡易化および他の定量法との比較
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The present paper deals with development of a practical fluorometric method for the determination of chlortetracycline (CTC) and with the reliability of the fluorometric method for CTC assay in ice as compared with several chemical and microbiologic assay methods. Two field fluorometric procedures have been developed in which the visual comparison of the fluorescence of a sample with that of a standard is made by using flat-bottom test tubes, on one hand, and a portable nephelo-colorimeter, on the other, viz., (1) by the use of test tubes: place 10-cc aliquots of supernatent of a thawed sample ice and the standard CTC solution series in a series of the flat-bottom test tubes of 27mm. in diameter, and then add to each test tube lcc. of the alum reagent; after a 5- to 10-minute standing, estimate the CTC concentration of the sample by comparing from top of the tubes the intensity of fluorescence of the sample with each of the standard series while being side-irradiated with a long wave ultra-violet ray lamp, (2) by the use of the portable nephelo-colorimeter: add 1-cc. aliquot of the alum reagent to 10-cc. aliquot of supernatent of a thawed sample ice and a standard CTC solution (4 or 2mcg./cc. depending on CTC content of the sample); after allowing it to stand for 10 minutes, place the sample in one of the comparison tube, a, and the standard in the other tube, b; set the bottom of an inner tube for the tube, b, at a depth of 30mm. and by adjusting the other inner tube find the reading, h, on the tube, a, which will give fluores-cence intensity comparable to that of the standard, both tubes being equally irradiated with the long wave ultra-violet lamp; the content of CTC (mcg./cc.) can be calculated by a formula, S×30/h, where, S, the CTC concentration of the standard solution employed. The feasibility of the fluorometric method for CTC-estimate in ice has been clearly shown by data that, while the fluorometric method gave a comparable value to that obtained by the microbiologic method, higher values were always resulted when assayed by a modified acid colorimetric8), the UPHAM et als7) and the SAKAGUCHI et als method9), but lower values by the Japan FDA method6).
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