本文選題：飼料 + 氨基酸　； 參考：《河北農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：飼料中的氨基酸是畜禽的重要營養(yǎng)物質(zhì),動物對蛋白質(zhì)的需求實(shí)際上是對氨基酸的需求。飼料中含有的氨基酸種類和含量是判定飼料質(zhì)量高低的重要指標(biāo)。因此,飼料中氨基酸的準(zhǔn)確分析、測定十分重要。但氨基酸分析誤差較大,可能是由于不同的水解條件和不同的分析方法所造成的。氨基酸的分析測定是一個復(fù)雜的過程,所以需要建立一套完善的飼料中氨基酸的檢測方法。蛋白質(zhì)的氨基酸分析分成三大步驟,第一,把蛋白質(zhì)徹底的水解為游離的氨基酸混合物;第二,用色譜的方法把混合的氨基酸分離成單個的氨基酸;第三,測定每個氨基酸的含量。目前,采用常規(guī)的酸水解法測定蛋白質(zhì)氨基酸的含量,因?yàn)樯彼嵩谒崴膺^程中幾乎都被破壞,無法與其他的氨基酸一起測定,所以只能單獨(dú)測定。本研究分為兩大部分,一部分是柱前衍生高效液相色譜法測定飼料中的氨基酸;另一部分是高效液相色譜法測定飼料中的色氨酸。具體內(nèi)容如下:本試驗(yàn)建立了以2,4-二硝基氯苯為柱前衍生劑的高效液相色譜法測定飼料中的氨基酸。通過對前處理?xiàng)l件中酸解劑、酸樣比、鹽酸濃度、水解溫度、水解時間、通氮?dú)�、微波水解時間、微波水解溫度、微波水解與常規(guī)水解的比較,衍生條件中衍生劑用量、衍生反應(yīng)時間、衍生反應(yīng)溫度、衍生反應(yīng)緩沖液、衍生反應(yīng)緩沖液p H?色譜條件中色譜柱、緩沖液、緩沖液p H?緩沖液濃度、三乙胺加入量、洗脫程序、流速、色譜柱溫度、有機(jī)洗脫溶劑、檢測波長進(jìn)行研宄,建立了19種氨基酸的標(biāo)準(zhǔn)曲線,其相關(guān)系數(shù)范圍為0.9901?0.9993;檢出限為0.69?5.70mg/L;加標(biāo)回收率為91.9%?110.7%。結(jié)果表明,本方法操作簡單,無干擾,具有較好的準(zhǔn)確度和較高的靈敏度,能夠滿足飼料中氨基酸的定量分析。本試驗(yàn)還建立了高效液相色譜法測定飼料中的色氨酸。通過對前處理?xiàng)l件中堿解劑種類、堿解劑濃度、堿解劑用量、堿解時間、堿解溫度、通氮?dú)?色譜條件中緩沖液、流動相p H?有機(jī)洗脫溶劑、流動相組成、流動相流速、色譜柱溫度、檢測波長的研宄,建立了色氨酸的標(biāo)準(zhǔn)曲線,其相關(guān)系數(shù)為0.9998;檢出限為O.llmg/L;加標(biāo)回收率為93.5%。結(jié)果表明,本方法操作簡單,無干擾,具有較好的準(zhǔn)確度和較高的靈敏度,能夠滿足飼料中氨基酸的定量分析。
[Abstract]:Amino acids in feed are important nutrients for livestock and poultry. The kinds and contents of amino acids in feed are important indexes to judge the quality of feed. Therefore, the accurate analysis and determination of amino acids in feed is very important. However, the error of amino acid analysis may be due to different hydrolysis conditions and different analytical methods. The analysis and determination of amino acids is a complex process, so it is necessary to establish a complete method for the determination of amino acids in feed. The amino acid analysis of proteins is divided into three steps: first, the complete hydrolysis of proteins into free amino acid mixtures; second, separation of mixed amino acids into single amino acids by chromatographic method; third, The content of each amino acid was determined. At present, the conventional acid hydrolysis method is used to determine the content of protein amino acids. Because tryptophan is destroyed in the process of acid hydrolysis, it can not be determined together with other amino acids, so it can only be determined separately. This study is divided into two parts, one is the determination of amino acids in feed by precolumn derivatization HPLC, and the other is the determination of tryptophan in feed by HPLC. The main contents are as follows: the determination of amino acids in feed by high performance liquid chromatography (HPLC) with 2o 4-dinitrochlorobenzene as precolumn derivative was established. By comparing the acid hydrolysis agent, acid sample ratio, hydrochloric acid concentration, hydrolysis temperature, hydrolysis time, nitrogen gas, microwave hydrolysis time, microwave hydrolysis temperature, microwave hydrolysis and conventional hydrolysis, the amount of derivative in the derivatization condition was compared. Derivatization reaction time, reaction temperature, buffer solution pH? In chromatographic conditions, the column, buffer solution pH? Buffer concentration, triethylamine addition, elution procedure, flow rate, chromatographic column temperature, organic elution solvent, detection wavelength were studied, and the standard curves of 19 amino acids were established. The range of correlation coefficient was 0.9901g / L 0.9993, the detection limit was 0.69 ~ 5.70 mg / L, and the recovery rate was 91.9% ~ 110.7. The results showed that the method was simple, without interference, had better accuracy and sensitivity, and could be used for quantitative analysis of amino acids in feed. The determination of tryptophan in feed by HPLC was also established. In this paper, the kinds of alkali hydrolysates, the concentration of alkali hydrolysates, the amount of alkali hydrolysates, the time of alkali hydrolysis, the temperature of alkali hydrolysis, nitrogen, the buffer solution in chromatographic conditions, the mobile phase pH? The standard curve of tryptophan was established by studying the organic elution solvent, the composition of mobile phase, the flow rate of mobile phase, the temperature of chromatographic column and the detection wavelength. The correlation coefficient was 0.9998, the detection limit was 0. Llmg / L, and the recovery rate was 93. 5%. The results showed that the method was simple, without interference, had better accuracy and sensitivity, and could be used for quantitative analysis of amino acids in feed.
【學(xué)位授予單位】：河北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S816.17

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本文編號：1988668