Vitamin K1 (VK1) has been widely used as a coumarin antagonist and for the treatment of hemorrhagic disease in veterinary practice. However, the potential mechanism of adverse reaction after VK1 injection has been not fully elucidated. In this study, two cases of anaphylactic reactions after subcutaneous VK1 injection were presented, and then an experimental study was performed to further characterize the anaphylactic reactions. Two dogs developed anaphylactic reactions after subcutaneous VK1 injections and were promptly treated with antihistamines and glucocorticoids, after which abnormal signs related to anaphylaxis disappeared. Subsequently, a study was undertaken to ascertain the nature of the adverse reactions to subcutaneous VK1 injection. Six healthy laboratory beagle dogs received subcutaneous VK1 administrations once daily for eight days. They were monitored for clinical signs after each injection, and blood samples were collected for the measurement of plasma histamine and immunoglobulin E concentrations using enzyme-linked immunosorbent assay. All six dogs showed mild angioedema after the VK1 injections. The dogs also displayed clinical signs including sneezing, coughing, skin reddening, excess salivation, pawing the ground, and somnolence on days 4, 6, and 8. Plasma histamine and immunoglobulin E concentrations were significantly increased by the repeated injections. In summary, this study describes anaphylactic reactions resulting from subcutaneous VK1 administration in dogs. Clinicians should be aware that the repeated subcutaneous injection of VK1 can trigger an anaphylactic reaction in dogs.
Vitamin K1 is a fat-soluble vitamin essential for human health. We used the Association of Official Analytical Chemists(AOAC) method 999.15, validated by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH) Guideline, to monitor vitamin K1 content in infant formula in the Republic of Korea. The Certified Reference Materials (CRM), NIST 1849a, were used for validation of the test method. The parameters of validation were linearity, range, specificity, accuracy, precision, LOD, LOQ and recovery. The linearity, R2 was above 0.999 and the analytical range was 0.001 to 0.22 μg/mL. The specificity was confirmed by the retention time (RT) and positive/negative sample test. The accuracy, precision and recovery values were 101.13 ± 0.13 %, 3.15 % and 81.60 % - 99.78 %. The LOD and LOQ values were 0.582 ng/mL and 1.76 ng/mL, respectively. Five livestock products inspection agencies voluntarily participated in the collaborative study. The HORRAT value was 0.6, which was within the reference value 0.5 – 2.0. The validated test method was applied for the monitoring of vitamin K1 in infant formula in the Republic of Korea. The results also conformed to 'The Standard for Livestock Products of Processing and Ingredient Specifications' and 'The Standard for Livestock Products Labeling'.
Vitamin K (phylloquinone) is an essential cofactor in the synthesis of active blood-clotting factors II, VII, IX and X. Deficiency of vitamin K leads to inadequate activity of these factors, resulting in bleeding. In this study, we investigated vitamin K1 content of agricultural products that are widely and specifically grown in Korea including 9 leaves and vegetables, 16 fruits, and 11 cereals and specialty crops. Vitamin K1 analysis of the agro-samples was by a validated, modified, reversed phase-HPLC method with fluorescence detection after post-column derivatization. The vitamin K1 content ranged from 1.83 to 682.73 μg/100 g in leaves and vegetables, 0.17 to 28.22 μg/100 g in fruits, and ND to 279 μg/100 g in cereals and specialty crops. Among the 36 samples, high content of vitamin K1 were found in Gugija (Lycium chinense Miller) leaves (average 682.73 μg/100 g) and Hansan ramie leaves (average 423.12 μg/100 g); however, mushroom, amaranth and Chinese artichoke showed no detectable levels. The results of ourstudy provide reliable vitamin K1 content of Korean grown agricultural products that expand nutritional information and food composition database.
돼지 수정란의 체외 생산 효율성 향상을 위해서는 배발생율과 더불어 고품질의 배를 조기에 선별해야 한다. 체외 배 발생율에 대한 보고는 많지만, 고품질의 배를 선별할 수 있는 기술에 대한 연구는 거의 없었다. 본 연구에서는 돼지 난포란 유래 수정란의 체외배양에 있어서 배반포로의 배 발달과 생존에 미치는 Vitamin K1(vit K1) 첨가 농도, 시기 및 시간의 효과를 검토하였다. 1.0 μM, 3.0 μM 및 6.0 μM vit K1을 배양 1일째 24시간 첨가한 결과, 배반포 발달율이 시험군이 14.5 ± 4.3, 0.0 및 0.0%로써 대조군의 35.5 ± 3.2%에 비하여 유의하게 낮았고(p<0.05), 배반포의 생존율도 대조군이 31.8 ± 2.6%로써 시험군의 22.2 ± 2.9, 0.0 및 0.0%에 비하여 유의하게 높았다(p<0.05). 상기 첨가 농도에서 첨가 시간을 달리한 결과, 1.0 μM 농도에서 6시간 처리군의 배반포 발달율과 생존율이 각각 26.5 ± 2.9% 및 47.2 ± 2.8%로써 가장 높았고 특히, 12시간 처리군보다 유의하게 높았다(p<0.05). 3.0 μM 농도에서는 대조군의 배발달율이 36.4 ± 3.1%로 가장 높았으나, 생존율은 3.0시간 첨가군이 41.7 ± 3.2%로 대조군에 비하여 유의하게 높았다(p<0.05). 6.0 μM 농도에서도 배발달율은 대조군(32.0 ± 2.8%), 생존율은 0.5시간 첨가군(42.9 ± 1.8%)이 가장 높았다. 각각의 vit K1 첨가 농도와 시간을 기준으로 서로 다른 배양 시기에 첨가한 결과, 1.0 μM 6시간 첨가군에서는 배반포 발달율은 배양 4일째 첨가군, 생존율은 배양 2일째 첨가군이 가장 높았다. 한편, 3.0 μM 3.0시간 및 6.0 μM 0.5시간 첨가군에서는 배양 4일째 첨가군의 배반포 발달율(59.5 ± 4.1% 및 50.0 ± 3.6%)과 생존율(72.7 ± 5.4% 및 79.2 ± 4.0%)이 대조군과 다른 시험군에 비하여 유의하게 높았다(p<0.05). 한편, vit K1 첨가에 따른 배반포의 세포 수를 조사한 결과, 첨가군(1.0 μM 6시간 배양 2일째, 3.0 μM 3.0시간 배양 4일째 및 6.0 μM 0.5시간 배양 6일째)이 53.4 ± 5.8, 49.4 ± 3.8 및 51.5 ± 4.5개로써 대조군의 40.2 ± 2.3개에 비하여 유의하게 많았다(p<0.05). 그러나 사멸세포 수는 시험군이 3.2 ± 0.9∼3.7 ± 2.1개로써 대조군의 4.2 ± 1.2개보다 적었으나, 유의차는 없었다. 세포 사멸 유도 유전자인 Bax mRNA 발현은 처리군과 대조군은 비슷하였으나, 세포 사멸 억제 유전자인 Bcl-xL mRNA 발현은 처리군이 대조군보다 높았고 특히, 6.0 μM 0.5시간 배양 4일째 첨가군이 가장 높았다. 이상의 결과로부터 돼지 미성숙 난포란 유래 수정란의 체외 배양에 vit K1의 첨가는 배반포의 생존율과 세포수 증가에 효과적이었다. 그 이유에 대해서는 아직 많은 부분이 밝혀져야 되겠지만, 고품질의 배반포 조기 선발에는 활용이 가능할 것으로 생각된다.