A simple and fast analytical method based on liquid chromatography-tandem mass spectrometry was developed for detection of the veterinary drugs acetanilide, anthranilic acid, antipyrine, cyproheptadine, diphenhydramine, DLmethylephedrine, and phenacetin in bovine milk. The target analytes were extracted from milk samples by using acetonitrile followed by clean-up with C18 and liquid-liquid purification with saturated n-hexane. A reverse-phase analytical column was employed with a mobile phase comprising (A) 0.1% formic acid in distilled water and (B) 0.1% formic acid in acetonitrile to achieve the best chromatographic separation. Matrix-matched calibration curves (r2 ≥ 0.9986) were constructed using six concentrations (1, 2, 5, 10, 20, and 40 μg/kg) of drugs in the milk matrix. Recoveries at three drug-spiking levels (5, 10, and 20 μg/kg) ranged from 71.2% to 103.8% with intra-day and inter-day relative standard deviation (RSD) values of ≤ 8.6%. The calculated limits of quantification (LOQ) were 0.19-7.1 μg/kg.

본 연구는 우유 중에서 덱사메타손을 효과적으로 정량 분석하기 위한 LC-MS/MS법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출 한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준용액을 이용하여 검량성을 작성한 결과, r2 > 0.999 이상의 직선성을 확인하였고, 덱사 메타손에 대한 검출한계와 정량한계는 각각 0.15와 0.5 ng/ mL이었다. 또한, 회수율은 98.9-109.6%로 나타났으며, 상대표준편차는 1.7-4.4%로 나타나 정확성이 우수하였으며, 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 우유 중 덱사메타손을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.

The current standard solutions for somatic cells used for calibration of electronic somatic cell counts as reference material in raw milk are preserved with bronopol, boric acid, sodium azide, or potassium dichromate, and have a shelf-life of only up to 6 days at 4 ± 2℃. In the present study, a set of somatic cell standard solutions (SCSS) with a stability of 5 months for calibration of electronic instruments was developed. Somatic cells collected from cow’s milk and stored in a bulk tank at a dairy plant were treated with 10% formaldehyde in order to improve stability, and then separated by centrifugation. The resulting somatic cell suspension was preserved with glycerin, thimerosal, and dimethyl sulfoxide, and diluted in 3% processed skim milk solution ranging from 200,000～250,000 (low level), 350,000～ 450,000 (medium level), and 550,000～650,000 (high level) cells/㎖. Each SCSS was verified by direct microscope somatic cell counting (DMSCC), C-reader, and commercial standard samples. The average somatic cell count determined by DMSCC was 248, 214, 226 × 103 cells/㎖, 436, 382, 420 × 103 cells/㎖, and 612, 595, 609 × 103 cells/㎖. The coefficient of variation representing the repeatability of DMSCC decreased as the number of cells increased, and was <10.0% in almost all SCSS samples (range 4.6～7.1%). No statistically significant difference in somatic cell concentration was observed after storage at refrigeration temperature (2～6℃) over a period of 22 weeks (5 months). The stabilized SCSS may be useful as a reference material for determination of somatic cell count and quality control in testing of bovine raw milk.

The present study was conducted to examine the effect of recombinant bovine somatotropin(γBST), which was administered to cow to promote milk production, on bST levels in milk. Fourteen cows were divided into two groups: 1) control cows received neither γbST nor vehicle, 2) treated cows were administered twice at two-week interval with 500 mg γbST each cow by subcutaneous injection. Milk samples were taken on day 0 (prior to injection), day 7 (7 days after 1st injection), day 21 (7 days after 2nd injection) and day 35 (21 days after 2nd injection). Milk bST concentration was measured by the radioimmunoassay method. There was no statistical difference(p$lt;0.05) in milk bST levels between two groups showing bST levels in the range of 1.8 ng/ml to 3.1 ng/ml. That is, γbST administration did not increase bST levels in milk.