人乳中B族维生素水平及其影响因素

发布时间：2019-04-13 20:40

【摘要】：研究背景和目的人乳为婴儿最佳食品,它能够提供0-6月龄婴儿几乎所有营养成分。人乳中B族维生素是纯母乳喂养婴儿摄入的唯一来源。人乳中B族维生素的水平将直接关系婴儿的健康。近年来越来越多的国内外学者关注人乳中B族维生素的水平。目前的研究涉及到影响人乳中B族维生素水平的因素主要有哺乳阶段、膳食因素、社会经济学因素、农村和城市等。人乳中B族维生素的研究方法方面,主要有微生物法、化学法、放射分析法、免疫分析法、HPLC法,2012年有人首次应用UPLC/MS-MS方法分析了人乳中5种游离B族维生素的水平。然而目前的研究仍存在一些不足,主要是乳样采集缺乏标准化(如任意次乳样或几毫升乳样)；样本量较小,缺乏代表性；分析方法通常一次只能测定一种维生素、前处理复杂、分析周期长、检测成本高；较少的研究测定了人乳中黄素腺嘌呤二核甘酸(FAD)的水平。因此开发一种高效、快速、准确的方法,进行大样本人乳中B族维生素水平的分析,将有助于了解人乳中B族维生素的水平及其影响因素,指导乳母膳食,改善婴儿健康状况,为修订婴儿B族维生素适宜摄入量提供科学依据。研究对象和方法通过优化质谱参数、分析柱、流动相、前处理方法等,建立了·UPLC-MS/MS法分析人乳中B族维生素水平,用回收率、准确性、精密度、检出限、线性范围、稳定性、基质效应等因素进行了方法学确认。从中国母乳成分数据库样品中抽样,抽取1778份母乳样品,采用UPLC-MS/MS法分析了1778份人乳样品的B族维生素水平,按不同哺乳阶段、20个调查点、全国11省、6大区域、南北地区、农村和城市、沿海和内陆进行了描述和归纳。描述不同乳母学历、经济水平、分娩方式、体格测量、膳食摄入指标下的人乳样品中的B族维生素水平,用两独立样本的t检验和单因素方差分析检验不同指标下人乳中B族维生素水平的差异；分析两因素间的相关性,通过线性回归方程筛选出人乳中的10种B族维生素水平各自的影响因素。研究结果UPLC-MS/MS方法前处理简单,线性范围拟合良好,准确度为98.4-108.0%,日内精密度和日间精密度分别为3.4-19.9%和5.9-18.1%、稳定性好,达到分析要求,适合大批量人乳样品多种B族维生素同时分析。初乳、过渡乳、成熟乳中B族维生素水平的中位数依次分别为：硫胺素(5.0μg/L、6.7μg/L、 27.2 μg/L),核黄素(29.3μg/L、41.5 μg/L、32.2μg/L),烟酸(464.2 μg/L、675.1 μg/L、635.7 μg/L)、维生素B6(4.7μg/L、18.8μg/L、70.3μg/L)、FAD (808.7 μg/L、1174.6μg/L、1023.9 μg/L)、泛酸(1770.9 μg/L、2628.3 μg/L、2119.4 μg/L)。目前分析的人乳中B族维生素水平在初乳阶段水平均是最低的,硫胺素和维生素B6水平随哺乳期增大而增大,核黄素、烟酸、FAD和泛酸水平随哺乳期增大均呈现先增大后减小的变化趋势。整个哺乳阶段里,农村乳母的硫胺素、核黄素和烟酸水平低于城市乳母,农村乳母的维生素B6水平高于城市乳母,城市乳母的FAD水平与农村乳母相近,农村乳母的泛酸水平在哺乳期0-30天里高于城市乳母,30天以后二者差异逐渐变小。沿海地区乳母乳汁的硫胺素、核黄素和烟酸水平高于内陆地区乳母,沿海地区乳母乳汁的FAD、泛酸和维生素B6水平低于内陆地区乳母。影响人乳中B族维生素水平的因素主要有哺乳阶段、膳食、地区、沿海和内陆、农村和城市、乳母学历、乳母职业、分娩方式。省份地区因素影响人乳中10种B族维生素水平；哺乳阶段影响人乳中除FAD以外的9种B族维生素水平,膳食因素影响人乳中除核黄素以外的9种B族维生素水平,乳母学历影响人乳中核黄素、泛酸、吡哆胺、生物素水平；乳母职业影响人乳中核黄素、泛酸水平；沿海和内陆地区影响人乳中硫胺素、吡哆醛、FAD、吡哆醇、吡哆胺水平；农村和城市影响人乳中尼克酰胺、吡哆醛、FAD、尼克酸、吡哆醇水平；分娩方式影响人乳中泛酸和吡哆胺水平。结论本研究建立了高效、快速的UPLC-MS/MS法分析人乳中10种B族维生素水平,检测方法具有良好的准确性和精密度；分析了全国1778份人乳样品,首次系统报道了全国20个调查点、11省份、6大区域、南北地区、农村和城市、沿海和内陆、不同哺乳阶段的人乳中B族维生素水平；研究发现人乳中B族维生素的影响因素主要包括哺乳阶段、膳食、地区、沿海和内陆、农村和城市、乳母学历、乳母职业、分娩方式。
[Abstract]:The background and purpose of the study are the best food for infants, which can provide almost all of the nutritional ingredients for infants from 0 to 6 months of age. Vitamin B in human milk is the only source of pure breast-fed infants. The level of B vitamins in human milk will directly relate to the health of the infant. In recent years, more and more domestic and foreign scholars are concerned about the level of B vitamins in human milk. At present, the factors affecting the level of vitamin B in human milk mainly include the feeding stage, the dietary factor, the social economic factor, the rural and the city, and so on. In the aspect of the study of B vitamins in human milk, the level of five free B vitamins in human milk was analyzed by the method of micro-organism, chemical method, radioanalysis, immunoassay, HPLC, and the first time in 2012 by UPLC/ MS-MS. However, there are still some deficiencies in the present study, such as the lack of standardization in the collection of milk samples (such as any sub-sample or a few ml of milk samples), the small sample size and the lack of representativeness; the analytical method can only measure one kind of vitamin at a time, the pretreatment is complicated, the analysis period is long, and the detection cost is high; The level of fAD in human milk was determined by a few studies. therefore, a high-efficiency, fast and accurate method is developed, and the analysis of the vitamin level of the B-family in the human milk of the large sample is carried out, So as to provide a scientific basis for revising the appropriate intake of vitamin B vitamins. The method of UPLC-MS/ MS was established to analyze the level of B vitamins in human milk by optimizing the mass spectrum parameters, the analytical column, the mobile phase and the pretreatment method. The recovery, the accuracy, the precision, the detection limit, the linear range and the stability of the human milk were determined by the method of UPLC-MS/ MS. The matrix effect and other factors have been confirmed by the methodology. 1778 human milk samples were sampled from Chinese milk component database samples, and 1778 human milk samples were analyzed by UPLC-MS/ MS. The levels of vitamin B in 1778 human milk samples were analyzed by UPLC-MS/ MS. The coastal and inland areas have been described and summarized. The vitamin levels of B vitamins in human milk samples were tested by t-test and single-factor analysis of variance of two independent samples, and the differences in vitamin levels of B-group in human milk were tested by t-test and single-factor analysis of variance of two independent samples. The correlation between two factors was analyzed, and the influence factors of 10 B vitamins in human milk were selected by linear regression equation. The method of UPLC-MS/ MS was simple, the linear range fit well, the accuracy was 98.4-108.0%, the intra-day precision and the day precision were 3.4-19.9% and 5.9-18.1%, and the stability was good. The median of the vitamin levels in the colostrum, the transitional milk and the mature milk were: thiamine (5.0. mu.g/ L, 6.7. mu.g/ L, 27.2. mu.g/ L), riboflavin (29.3. mu.g/ L, 41.5. mu.g/ L, 32.2. mu.g/ L), nicotinic acid (464.2. mu.g/ L, 675.1. mu.g/ L, 635.7. mu.g/ L), vitamin B6 (4.7. mu.g/ L, 18.8. mu.g/ L, 70.3. mu.g/ L), FAD (808.7. mu.g/ L, 1174.6. mu.g/ L, 1023.9. mu.g/ L), pantothenic acid (1770.9. mu.g/ L, 2628.3. mu.g/ L, 2119.4. mu.g/ L). At present, the level of B vitamins in human milk is the lowest in the phase of the colostrum, and the levels of thiamine and vitamin B6 increase with the increase of the lactation period, and the levels of riboflavin, niacin, FAD and pantothenic acid increase with the increase of the lactation period. The level of thiamine, riboflavin and niacin in the rural lactating mother is lower than that of the city's milk mother in the whole lactation period, and the level of the vitamin B6 of the dairy mother in the rural area is higher than that of the urban milk mother, and the FAD level of the urban milk mother is similar to that of the rural dairy mother. The level of pantothenic acid in the rural lactating women is higher than that of the city's milk mother in the period 0-30 days of the lactation period, and the difference between them gradually becomes smaller after 30 days. The levels of thiamine, riboflavin and niacin in the breast milk of the coastal area are higher than that in the inland areas, and the levels of FAD, pantothenic acid and vitamin B6 in the milk in the coastal area are lower than that in the inland areas. The factors affecting the level of vitamin B in human milk mainly include the feeding stage, the diet, the area, the coastal and the inland, the rural and the city, the mother's degree, the mother's occupation and the mode of delivery. The factors of the province affect the level of 10 B vitamins in human milk, and the level of vitamin B vitamins except FAD in human milk is affected by the feeding stage, and the dietary factors affect the levels of vitamin B vitamins except for riboflavin in human milk, and the milk mother's degree affects the riboflavin and pantothenic acid in human milk. in that coastal and inland areas, the human milk, such as thiamine, calcitral, fad, trialdol, and alammine, are present in the coastal and inland areas; and in the rural and urban areas, the human milk, such as nicholamide, fad, fad, nicotinic acid, The levels of pantothenol in human milk were affected by delivery, and the level of pantothenic acid in human milk was affected by delivery. Conclusion The present study established an efficient and rapid UPLC-MS/ MS method for the analysis of 10 B-family vitamin levels in human milk. The detection method has good accuracy and precision.1778 human milk samples were analyzed. The first system reported 20 investigation points,11 provinces,6 large areas and north and south areas. The level of vitamin B in human milk in rural and urban, coastal and inland and different lactation stages is studied. The factors affecting the vitamin in human milk mainly include the feeding stage, the diet, the area, the coastal and the inland, the rural and the city, the mother's degree, the mother's occupation and the mode of delivery.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R155.5;O657.63

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