Study of the Impact of Light Through the Vitamin $B_{12}$/Folate Inspection

Vitamin $B_{12}$/Folate 검사 시 빛의 영향에 대한 고찰

  • Cho, Eun Bit (Department of Nuclear Medicine, Asan Medical Center) ;
  • Pack, Song Ran (Department of Nuclear Medicine, Asan Medical Center) ;
  • Kim, Whe Jung (Department of Nuclear Medicine, Asan Medical Center) ;
  • Kim, Seong Ho (Department of Nuclear Medicine, Asan Medical Center) ;
  • Yoo, Seon Hee (Department of Nuclear Medicine, Asan Medical Center)
  • Received : 2012.08.31
  • Accepted : 2012.10.05
  • Published : 2012.10.27

Abstract

Purpose : Vitamin $B_{12}$ and Folate are for anemia work-up which is well known for its sensitivity of light; the screening manual also specifies to be careful with light conditions. According to this, our laboratory minimized the exposure of light when inspecting the Vitamin $B_{12}$ and Folate, but the exposure cannot be wholly blocked due to other various factors such as when conducting specimen segregation. Thus, this inspection is to identify to what extent light can influence and whether the exclusion of light is mandatory during the Vitamin $B_{12}$/Folate test. Materials and Methods : We have conducted two experiments of identifying the extent of light's influence when conducting the Vitamin $B_{12}$/Folate test and also when specimens are under preservation. These experiments were progressed with various concentrations of patients' specimens which were requested to our hospital in March 2012. The first experiment is to verify the results on Vitamin $B_{12}$/Folate dependent on light exposure during the experiment. In the process, we have compared the results of light exposure/exclusion during the incubation process after the reagent division. The second experiment is about the impact of light exposure on the results on Vitamin $B_{12}$/Folate during the preservation. For 1, 2, 7 days the light on the specimen were wholly blocked and were preserved under $-15^{\circ}C$ temperature refrigeration. Then, we compared the results of light-excluded specimen and the exposed one. Results : When conducting first experiment, there were no noticeable changes in the Standard and specimen's cpm, but for Vitamin $B_{12}$, the average result of specimen exposed to light increased 7.8% compare to that of excluded one's. Furthermore, in the significant level 0.05, the significance probability or the p-value was 0.251 which means it has no impact. For Folate, the result being exposed to light decreased 5.4%, the significance probability was 0.033 which means it has little impact. For the second preservation, the result was dependent on the light exposure. The first day of preservation of Vitamin $B_{12}$, the clinical material exposed to light was 11.6%, second day clinical material exposed to light was 10.8%, seventh day clinical material exposed to light increased 3.8%, the significance probability of the $1^{st}$, $2^{nd}$, $7^{th}$ day is 0.372, 0.033, 0.144 respectively, and which indicates that the $1^{st}$ and $7^{th}$ day seems to have no impact. For Folate's case, the clinical material exposed to light has increased 1.4% but hardly had impact, $2^{nd}$ day clinical material being exposed to light was 6.1%, $7^{Th}$ day clinical material being exposed to light decreased 5.2%. The significance probability of Folate on the $1^{st}$, $2^{nd}$, $7^{th}$ day is 0.378, 0.037, 0.217 respectively, and the $1^{st}$ day and the $7^{th}$ day seems to have no impact. Conclusion : After scrutinizing the impact of light exposure/exclusion, Vitamin $B_{12}$ has no impact, while Folate seems to have no noticeable influence but light exclusion is recommended due to its significance probability of 0.033 when conducting experiment. During the preservation, the $2^{nd}$ day result depend on the light exclusion seems to have impact or influence. However, to consider the complication of the experimental process, the experiment including technical errors is predictable. Hence, it is likely to have no impact of light. Nevertheless, it is recommendable to exclude the light during the long preservation as per the significance probability (p-value) of $1^{st}$ and $7^{th}$ day has been diminished.

Vitamin $B_{12}$, Folate는 아미노산과 핵산의 합성에 필수적인 영양소로 DNA 복제에 관여하는 효소이다. 체내에서는 신체의 성장에 활발하게 관여하여 세포분열과 증식을 하는 조혈조직에 대한 영향이 커 빈혈의 감별진단 검사의 하나로서 측정된다. 이런 Vitamin $B_{12}$, Folate는 빛에 민감한 검사로 알려져 있어 검사 매뉴얼 또한 검사 시 실험 시, 보관 시 모두 빛의 노출을 피하도록 하고 있다. 이에 따라 본원의 검사실에서는 Vitamin $B_{12}$, Folate 검사시 빛의 노출을 최소한으로 하고 있다. 하지만 검체 분리 보관 지연 및 검사 시 다른 여러 가지 요인들로 인해 빛의 영향을 완전히 배제할 수는 없다. 따라서 Vitamin $B_{12}$, Folate 검사 시 빛의 영향 정도와 빛 차단이 필수적인 지의 여부를 확인하고자 실험 시 빛의 영향 정도와 실험 전 보관 시 빛의 영향 정도로 나누어서 분석해보고자 한다. 2012년 3월, 본원에 의뢰된 다양한 농도의 환자 검체 10개를 대상으로 실험시 빛의 영향과 검체 보관 시 빛의 영향 정도를 확인하는 두 가지 실험을 실시하였다. 첫 번째는 Vitamin $B_{12}$, Folate 실험시 반응 단계에서 빛을 차단한 결과와 빛에 노출된 결과를 비교하였다. 두 번째는 Vitamin $B_{12}$, Folate 실험 전 검체 보관 시 1, 2, 7일 동안 빛을 차단하여 보관한 결과와 빛에 노출된 결과를 비교하였다. 첫 번째, 실험 시 빛 차단의 여부에 따른 검사 결과 Vitamin $B_{12}$의 경우 영향이 없고 Folate의 경우 큰 영향은 없지만 유의확률이 0.033(상관식 y=1.02x-0.86)으로 실험 시 빛을 차단하는 것을 권장한다. 두 번째, 보관시 빛 차단의 여부에 따른 검사 결과 1일, 7일에서는 영향이 없고 2일에서는 Vitamin $B_{12}$ 유의확률이 0.033, Folate 유의확률이 0.037로 영향이 있었다. 이는 복잡한 검사 방법으로 인한 기술적 오류가 포함되어 있을 것으로 예상되어 1, 2, 7일 모두 보관 시 빛에 영향이 없는 것으로 사료된다. 그러나 1일차에 비해 7일차 유의확률 p-value가 감소함에 따라 장기간 보관 시 빛을 차단할 것을 권장한다.

Keywords