구연산이 Leucomalachite green 시약의 안정성에 미치는 영향
Effect of Citric acid on the Stability of Leucomalachite green reagents
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범죄 현장에서 혈흔을 찾기 위해서 다양한 종류의 시약들이 사용되고 있는데 leucomalachite green(LMG) 도 그중 하나이다. LMG 시약은 타 시약에 비해 특이성이 높다고 알려져 있지만, 상온에서 발색이 빨라 사건현장에서 이용하는 것이 어렵다. 본 연구에서는 초산 대신 항산화 효과를 나타내는 구연산으로 LMG 시약(NF-LMG)을 제조한 후, 민감도, 특이성 그리고 안정성 측면에서 영향을 평가하였다. 분광광도법을 통해 발색된 시약의 최대 흡수 파장인 620 nm를 확인하였고, 안정성과 민감도 평가에 활용하였다. NF-LMG 시약은 특이성 측면에선 LMG 시약과 유사했지만, 희석 혈흔 시료와 반응할 때 발색 강도는 낮아져 민감도는 약간 감소했다. NF-LMG 시약의 민감도를 개선하기 위해서 구연산 나트륨을 첨가해 시약의 pH를 높일 수 있지만, pH 상승은 보관 중 안정성을 떨어뜨리는 결과를 초래했다. LMG 시약과 NF-LMG 시약 모두에서 보관 온도의 상승은 시약의 안정성을 급격히 떨어뜨리는 결과를 나타냈지만, 동일 온도 조건에선 NF-LMG 시약이 LMG 시약보다 50배 정도의 보관 안정성이 높았다. 실험 결과를 통해, 구연산의 항산화 효과를 확인하였고, NF-LMG 시약이 사건 현장에 적합하다는 사실을 알 수 있었다.
Abstract
Various types of presumptive regents are used to find bloodstains at crime scenes, and leucomalachite green(LMG) is one of them. Despite the high specificity of the LMG reagent, it is not easy to use in the field because the color develops quickly at room temperature. In this study, LMG reagent was prepared with citric acid(NF-LMG), which has an antioxidant effect, instead of acetic acid, its effects were evaluated in terms of sensitivity, specificity, and stability. Using the spectrophotometric method, it was confirmed that the maximum absorption wavelength of the colored reagent was 620 nm, and it was used to evaluate stability or sensitivity through color development during storage. The specificity of the NF-LMG reagent was similar to the LMG reagent, but the sensitivity was thought to be lowered because the color intensity decreased after reacting with bloodstains. To improve the sensitivity of the NF-LMG reagent, the pH of the reagent can be increased by adding sodium citrate, but the higher the pH, the lower the stability of the reagent. As the storage temperature increased, the stability of the LMG reagent lowered. However, at the same storage temperature, the stability of NF-LMG reagent was about 50 times higher than that of the LMG reagent. Based of the experimetal results, it is believed that LMG reagent prepared with citric acid is suitable for the crime scene.
참고문헌 (14)
- 이민지, "오래된 혈흔 시료에 대한 혈청학적 식별법의 비교 연구", 분석과학회지, 31(5), pp.201~207 (2018)
- 오유리, "혈흔 및 정액흔 검사를 위한 PeroxtesmoⓇ KM 및 PhosphatesmoⓇ KM 페이퍼의 유효성 검토", Journal of Scientific Criminal Investigation, 11(1), pp.18~24 (2017)
- Budowle B, "The presumptive reagent fluorescein for detection or dilute bloodstains and subsequent STR typing of recovered DNA", Journal of Forensic Science, 45(5), pp.1090~1092 (2000)
- Culliford BJ, "The benzidine test: A critical review", Journal of Forensic Science, 9(1), pp.175~191 (1964)
- Tobe SS, "Evaluation of six presumptive tests for blood their specificity, sensitivity and effect on high molecular-weight DNA", Journal of Forensic Science, 52(1), pp.102~109 (2007)
- Cox M, "A study of the sensitivity and specificity of four presumptive tests for blood", Journal of Forensic Science, 36(5), pp.1503~1511 (1991)
- Ahn SH, "Accurate determination of malachite green and leucomalachite green on fish using isotope dilution chromatography", mass spectrometry(ID-LC, Bulletin of the Korean Chemical Society, pp.2010, 31(11)[DOI]
- Ahlquist DA, "Use of leuco-dyes in the quantitative colorimetric microdetermination of hemoglobin and other heme compounds", Clinical Chemistry, 21(3), pp.362~369 (1975)
- Luiz FA, "Direct determination of malachite green and leucomalachite green in natural waters by exploiting solid-phase sorption and digital image", Orbital: The Electronic Journal of Chemistry, 12(4), pp.193~204 (2020)
- Tiwari KK, "A new spectrophotometric method for the determination of ascorbic acid using leucomalachite green", Journal of Chinese chemical society, 57, pp.105~110 (2010)
- Evan MR, "Antioxidant properties of citric acid interfere with the uricase based measurement of circulating uric acid", Journal of Pharmaceutical and Biomedical Analysis, 164(5), pp.460~466 (2019)
- Syed AA, "New reagents for detection of faecal occult blood", J pharmaceutical and biomedical analysis, 24, pp.581~586 (2001)
- Taniquchi Y, "Method for stabilizing leuco dye", US patent 20070026523A1 Feb 1, ~ (2007)
- Taniquchi Y, "Method for stabilizing oxidizable color development reagent", US patent 20080241816A1 Oct 2, ~ (2008)