Edinboro Leslie E, Karnes H Thomas
Department of Pharmaceutics, Virginia Commonwealth University, School of Pharmacy, P.O. Box 980533, 410 N 12th Street, Richmond, VA 23298-0533, USA.
J Chromatogr A. 2005 Aug 12;1083(1-2):127-32. doi: 10.1016/j.chroma.2005.06.032.
Aflatoxins produced by food-borne molds are known carcinogenic toxins. Aflatoxin B1 (AFB1) is reported as the most toxic of this class of mycotoxins. We have coupled immunoaffinity column extraction with LC/MS to produce a sensitive and selective approach for the study of AFB1. As AFB1 can be potentially found in tobacco it is of interest to establish whether AFB1 can be transferred from a cigarette fortified with AFB1, to the sidestream smoke. Previous studies have found that AFB1 does not transfer to the mainstream smoke. Since sidestream smoke may contain higher concentrations of some smoke components, a method was developed to analyze the sidestream smoke produced from machine-smoked cigarettes. Sidestream smoke condensates collected on Cambridge filter pads were extracted with isopropanol, then further purified using immunoaffinity extraction columns. The extracts were then analyzed by LC/MS and LC/MS/MS. An instrumental limit of detection (LOD) was established at 3.75 pg injected on column, with the limit of quantitation (LOQ) equal to 11.25 pg on column for both LC/MS and LC/MS/MS. The instrument was found to be linear from 11.25 pg to 150 pg (r > 0.995.) Precision ranged from 4.2% to 8.4% at the LOQ, while accuracy ranged from 0.53% to 1.33%. The immunoaffinity extraction method LOD was determined to be 100 pg fortified onto the Cambridge filter. The LOQ was 350 pg. The average recovery of the AFB1 from the Cambridge pad was 82.9% over the range of 100-1000 pg fortified onto the pad. AFB1 was not detected in unfortified cigarettes. A transfer experiment, fortifying cigarettes at 1 microg/cigarette determined that AFB1 was transferred only slightly from the burning cigarette to the sidestream smoke. The mean percent transfer was 0.087%.
由食源性霉菌产生的黄曲霉毒素是已知的致癌毒素。黄曲霉毒素B1（AFB1）据报道是这类霉菌毒素中毒性最强的。我们将免疫亲和柱萃取与液相色谱/质谱联用，开发出一种灵敏且具选择性的方法来研究AFB1。由于AFB1可能存在于烟草中，因此确定AFB1是否能从添加了AFB1的香烟转移到侧流烟雾中很有意义。先前的研究发现AFB1不会转移到主流烟雾中。鉴于侧流烟雾可能含有更高浓度的某些烟雾成分，于是开发了一种方法来分析机器抽吸香烟产生的侧流烟雾。收集在剑桥滤片上的侧流烟雾冷凝物先用异丙醇萃取，然后使用免疫亲和萃取柱进一步纯化。萃取物随后通过液相色谱/质谱和液相色谱/质谱/质谱进行分析。仪器检测限（LOD）确定为柱上进样3.75 pg，液相色谱/质谱和液相色谱/质谱/质谱的定量限（LOQ）均为柱上11.25 pg。该仪器在11.25 pg至150 pg范围内呈线性（r > 0.995）。定量限处的精密度为4.2%至8.4%，而准确度为0.53%至1.33%。免疫亲和萃取法的检测限确定为添加到剑桥滤片上的100 pg。定量限为350 pg。在添加到滤片上的100 - 1000 pg范围内，AFB1从剑桥滤片的平均回收率为82.9%。未添加AFB1的香烟中未检测到AFB1。一项转移实验，以每支香烟1微克的量添加AFB1，结果表明AFB1仅从燃烧的香烟略微转移到侧流烟雾中。平均转移率为0.087%。
