Gatautis V J, Pearson K H
Clin Chim Acta. 1987 Jul 15;166(2-3):195-206. doi: 10.1016/0009-8981(87)90422-0.
A method was developed for the extraction and separation of human plasma carotenoids and quantitation of beta-carotene. Carotenoids were extracted from plasma with ethanol: hexane and separated by C18 reversed phase HPLC using spherical 3 micron packing. beta-Carotene was identified and quantitated using an external standard. The within-run precision of three different plasma pools ranged from 3.53-5.72% relative standard deviation (RSD). The between-run precision was 7.34% RSD. The method was linear to 500 micrograms/l with a statistical detection limit of 3.80 micrograms/l. Recovery of added beta-carotene was from 90.41-100.37%. This method was compared to a spectrophotometric 'total carotene' method. The mean plasma concentrations of 25 male and 25 female human volunteers for the 'total carotene' were 1,549 micrograms/l for all samples, 1,487 micrograms/l for males and 1,611 micrograms/l for females. The corresponding true beta-carotene concentrations obtained by HPLC analysis were 134.8, 115.9 and 153.7 micrograms/l, respectively. The true beta-carotene concentrations were on the average only 8.76% (8.07% for males and 9.46% for females) of the concentrations obtained by the spectrophotometric 'total carotene' method. Correlation between the methods had an r = 0.6107. The poor correlation is due to the difference in the measured components. Total carotene methods measure all solvent extractable moieties having absorbance in the 430-460 nm region, while the HPLC method quantitates true beta-carotene after chromatographic separation from other carotenoids. Reference intervals were established for plasma beta-carotene using REFVAL, an IFCC computer program for determining statistical reference intervals. The reference interval for all samples is 40 to 344 micrograms/l.
开发了一种用于提取和分离人血浆类胡萝卜素并定量β-胡萝卜素的方法。类胡萝卜素用乙醇：己烷从血浆中提取，并用球形3微米填料的C18反相高效液相色谱法分离。β-胡萝卜素使用外标进行鉴定和定量。三种不同血浆池的批内精密度相对标准偏差（RSD）为3.53 - 5.72%。批间精密度为7.34%RSD。该方法在500微克/升范围内呈线性，统计检测限为3.80微克/升。添加的β-胡萝卜素回收率为90.41 - 100.37%。将该方法与分光光度法“总类胡萝卜素”方法进行了比较。25名男性和25名女性人类志愿者的“总类胡萝卜素”血浆平均浓度，所有样本为1549微克/升，男性为1487微克/升，女性为1611微克/升。通过高效液相色谱分析获得的相应真实β-胡萝卜素浓度分别为134.8、115.9和153.7微克/升。真实β-胡萝卜素浓度平均仅为分光光度法“总类胡萝卜素”方法所获浓度的8.76%（男性为8.07%，女性为9.46%）。两种方法之间的相关性r = 0.6107。相关性较差是由于所测成分不同。总类胡萝卜素方法测量在430 - 460纳米区域有吸光度的所有可溶剂萃取部分，而高效液相色谱法在与其他类胡萝卜素进行色谱分离后对真实β-胡萝卜素进行定量。使用REFVAL（一种用于确定统计参考区间的国际临床化学和检验医学联合会计算机程序）建立了血浆β-胡萝卜素的参考区间。所有样本的参考区间为40至344微克/升。
