凱氏定氮儀和發(fā)展史課件

1、凱氏定氮一個(gè)使用了120年的依舊年輕的技術(shù)BCHI Labortechnik AGKatharina JehleFlawil, SwitzerlandJuly 05/KJ1China_Kjeldahl Solutions凱氏定氮July 05/KJ1China_Kjeldahl 主題：蛋白的基本概念 凱氏定氮的方法 方法的發(fā)展 Bchis 消化單元 Bchis消化單元Kjeldahl法問題解決July 05/KJ2China_Kjeldahl Solutions主題：蛋白的基本概念July 05/KJ2China_Kje蛋白的基本概念Basics ProteinJuly 05/KJ3China

2、_Kjeldahl Solutions蛋白的基本概念Basics ProteinJuly 05/K為什么要分析蛋白?蛋白質(zhì)是： Basic nutrient基本的營(yíng)養(yǎng) Impact on caloric value (17 kJ resp. 4 kcal)熱量來(lái)源 Parameter for quality (e.g. meat)品質(zhì)參數(shù) Influence on production process (e.g gelation, extrusion)影響產(chǎn)品的中間處理（例，凍結(jié)，壓榨） Influence on flavour and colour of food (e.g. baking

3、process bread, roasting process coffee)影響食品的 風(fēng)味和顏色July 05/KJ4China_Kjeldahl Solutions為什么要分析蛋白? Basic nutrient基本的營(yíng)養(yǎng)蛋白的基本結(jié)構(gòu)是氨基酸，分子式如下： There are about 200 amino acids found in nature 自然界已發(fā)現(xiàn)的大概有200種氨基酸 There are about 20 amino acids in a protein hydrolysate 蛋白水解后大概有20種氨基酸Basics ProteinJuly 05/KJ5China_

4、Kjeldahl Solutions蛋白的基本結(jié)構(gòu)是氨基酸，分子式如下： There are a怎么測(cè)定蛋白質(zhì)？通過檢測(cè)氮的含量計(jì)算蛋白含量The nitrogen content in protein is in a small range (15% - 18%, mean: 16%)蛋白中氮的含量在1518波動(dòng)，平均16Protein is determined by analysing the nitrogen content and calculating the protein 通過氮含量計(jì)算蛋白含量Basics ProteinJuly 05/KJ6China_Kjeldahl So

5、lutions怎么測(cè)定蛋白質(zhì)？The nitrogen content i凱氏定氮概述July 05/KJ7China_Kjeldahl Solutions凱氏定氮概述July 05/KJ7China_Kjeldah誰(shuí)是凱氏?Johan Kjeldahl 1849 19001849 born in Denmark graduation from university Scientist at Carlsberg Breweries Fermentation studies Development of a method for exact determi- nation of nitrogen

6、 in animal and vegetable material Research on carbohydrates in barley and malt1883 Publication of the Kjeldahl methodJuly 05/KJ8China_Kjeldahl Solutions誰(shuí)是凱氏?Johan Kjeldahl 1849 190凱氏定氮的原理1. Digestion消化Conversion of Protein-Nitrogen to NH4+ 將蛋白中氮轉(zhuǎn)化為NH4+ 2. Distillation蒸餾分離收集NH3Separation of NH33. Tit

7、ration滴定用酸中和滴定Measure of the Amount of Acid that was neutralised by NH3 July 05/KJ9China_Kjeldahl Solutions凱氏定氮的原理1. 2. 3. July 05/KJ9ChiDigestion消化 (I)BlankSample1.0722 tabs ofCatalystSulphuric acidWeigh sample稱重 1 g of dry organic material (homogeneous) 1克干的有機(jī)物質(zhì)（均質(zhì)）Sample Blank 空白 add everything e

8、xcept sampleAdd reagents加試劑 10 g or 2 tabs of Kjeldahl catalyst mixture 10g 或2片凱氏消化片20ml 硫酸 樣品均質(zhì)，稱重后加入試劑July 05/KJ10China_Kjeldahl SolutionsDigestion消化 (I)BlankSample1.07Digestion消化 (II)vapours to ScrubberDigestion digest at 370 C for 90 min or until the mixture is clear (green / blue color) + 30 mi

9、n 370 90 mins 或 消化澄清 后30分鐘 Digestion July 05/KJ11China_Kjeldahl SolutionsDigestion消化 (II)vapours to DigDistillation 蒸餾(I)manually or automatedDilution dilute the cooled digestion solution with 50 ml deionised water加入50ml 去離子水稀釋Dilution稀釋July 05/KJ12China_Kjeldahl SolutionsDistillation 蒸餾(I)manually

10、Distillation蒸餾 (II)Boric acid titration硼酸滴定 add 60 ml of Boric acid 4% 加 460ml 的硼酸 add 2 to 3 drops of indicator according to Sher 加兩至3滴的指示劑Back titration反滴定 add 20.0 ml of sulphuric acid standard solution (0.5 N or equivalent)加20ml的硫酸標(biāo)準(zhǔn)液（0.5N) add 2 to 3 drops of indicator according to Sher加2到3滴指示劑

11、Preparation of Receiver 準(zhǔn)備接受液July 05/KJ13China_Kjeldahl SolutionsDistillation蒸餾 (II)Boric acid Distillation蒸餾 (III)Sodium hydroxide NaOH add 90 ml of NaOH 32% to the diluted digestion solution加90ml 32的NaOH到溶液中Distillation蒸餾 distill for 4 min.蒸餾4分鐘 Neutralisation and DistillationJuly 05/KJ14China_Kje

12、ldahl SolutionsDistillation蒸餾 (III)Sodium hydTitration滴定7.15Boric acid titration 硼酸滴定 titrate the condensate with sulphuric acid standard solution (0.5 N or equivalent) to the end point of pH 4.65 用0.5N的硫酸標(biāo)準(zhǔn)液中和滴定至 終點(diǎn)PH4.65Back titration 反滴定 titrate the condensate with sodium hydroxide standard solut

13、ion (0.5 N or equivalent) 用0.5N的NaOH標(biāo)準(zhǔn)液中和滴定 TitrationJuly 05/KJ15China_Kjeldahl SolutionsTitration滴定7.15Boric acid titrCalculation計(jì)算% P =(Consumption-Blank) x 1.4007 x N x PF x 100Sample Size1.4007:1ml 0.1N Volumetric Solution 1.4007mg NN:Normality AcidPF: Protein factor N Protein July 05/KJ16China_

14、Kjeldahl SolutionsCalculation計(jì)算% P =(ConsumptioProtein Factors蛋白轉(zhuǎn)換因子Sample typeProtein FactorGelatine5.55Dairy products6.38Meat6.25Fish6.25Nuts, oilseeds5.4Cereals6.25Fruit, Vegetable6.25Corn6.25Eggs6.25July 05/KJ17China_Kjeldahl SolutionsProtein Factors蛋白轉(zhuǎn)換因子Sample ty凱氏定氮法的發(fā)展July 05/KJ18China_Kjeld

15、ahl Solutions凱氏定氮法的發(fā)展July 05/KJ18China_Kjel最早July 05/KJ19China_Kjeldahl Solutions最早July 05/KJ19China_Kjeldahl Buchi公司開始凱氏分析于：1961開始生產(chǎn)上市凱氏定氮儀市場(chǎng)上第1個(gè)帶有內(nèi)置蒸汽發(fā)生器的蒸餾儀，大大提高蒸餾效率July 05/KJ20China_Kjeldahl SolutionsBuchi公司開始凱氏分析于：1961July 05/KJ2定氮儀在 Bchi的發(fā)展演變July 05/KJ21China_Kjeldahl Solutions定氮儀在 Bchi的發(fā)展演變Ju

16、ly 05/KJ21ChiBchi 今日Digestion Units and Scrubber 消化單元和排廢裝置Distillation Units蒸餾單元July 05/KJ22China_Kjeldahl SolutionsBchi 今日Digestion Units and ScBchis Digestion Units 消化單元July 05/KJ23China_Kjeldahl SolutionsBchis Digestion Units 消化單元Ju消化加熱方式Infrared Heating紅外加熱Block Heating整體鋁塊加熱July 05/KJ24China_Kj

17、eldahl Solutions消化加熱方式Infrared HeatingBlock He紅外消化爐特點(diǎn) 紅外加熱，通過輻射方式，非接觸式加熱 升溫速度快（10min預(yù)熱） 可以升溫至650C消化管排廢和消化管緊密聯(lián)接 6 管消化儀(K-424) 或12 管 (K-435) 加熱步驟可控，定時(shí)器 12 管 (K-435)K-424K-435July 05/KJ25China_Kjeldahl Solutions紅外消化爐特點(diǎn) 紅外加熱，通過輻射方式，非接觸式加熱 K-4整體鋁塊加熱消化爐的特點(diǎn)K-437K-438 鋁塊加熱 - 接觸式加熱 可以升溫到450C 溫度時(shí)間可控消化管排廢和消化管緊

18、密聯(lián)接 酸氣回收裝置可控 20個(gè)消化位置 自動(dòng)提升 (K-438 only) K-438: 程序化 (時(shí)間延遲, 升溫梯度, 自動(dòng)停止,)July 05/KJ26China_Kjeldahl Solutions整體鋁塊加熱消化爐的特點(diǎn)K-437K-438 鋁塊加熱JulDigestion紅外和整體鋁塊消化爐的區(qū)別 Block鋁塊 IR紅外預(yù)熱時(shí)間 60 min 5 min溫控Temperature easy difficultcontrol加熱單元最大加熱溫度 450C 600CJuly 05/KJ27China_Kjeldahl SolutionsDigestion紅外和整體鋁塊消化爐的區(qū)別

19、July 05/Digestion 鋁塊Block 紅外IR Water samples + +up to 100 mlLiquid samples + +up to 15 mlDry organic material + +0.2 g to 5 gHumid organic material + +0.2 g to 5 gFoamy samples + +Mineral oil + +Differences IR Block 紅外和整體鋁塊消化爐的區(qū)別July 05/KJ28China_Kjeldahl SolutionsDigestion 鋁塊Block 紅外IR如何處理消化過程中的酸霧？

20、Sulphuric acid vapours are corrosive and vitriolic酸霧具有腐蝕性和毒性，解決方法： 1) 消化爐排廢裝置，標(biāo)準(zhǔn)配置 2) 中和酸霧的廢氣循環(huán)回收裝置July 05/KJ29China_Kjeldahl Solutions如何處理消化過程中的酸霧？Sulphuric acid vaScrubber B-414模塊化設(shè)計(jì), 可達(dá)4步高效能泵系統(tǒng), 34 l/min大的回收試劑腔, 3 l大的氣液接觸面積, 提高操作性能可加NaOH or 碳酸鈉 (最高 NaOH 20% 但推薦濃度 NaOH 10%)可以選擇抗酸，提供特殊密封墊（抗酸）吸收步驟使用

21、活性炭吸附July 05/KJ30China_Kjeldahl SolutionsScrubber B-414模塊化設(shè)計(jì), 可達(dá)4步July Step 1 (optional)冷凝 Step 2 (standard)中和Step 3 (standard)吸收Step 4 (optional) 反應(yīng)Scrubber B-414July 05/KJ31China_Kjeldahl SolutionsStep 1 (optional)Step 2 (stand消化爐特殊附件Suction module for aqueous samples水類樣品吸收模塊帶有冷凝漕的吸收模塊，專門處理水類樣品，防止

22、交叉污染用于H2O2 消化的吸收模塊 (僅適用于 K-424 and K-435) 用于連續(xù)添加 H2O2 的漏斗 環(huán)保 (不需要催化劑) 更短的消化時(shí)間 減少起沫July 05/KJ32China_Kjeldahl Solutions消化爐特殊附件Suction module for aqueReflux module 回流模塊(僅適用于 K-424 and K-435)適用于必須在回流狀況下才能進(jìn)行的消化Module for Digestionswhich have to be done underreflux conditionsDigestion Units: Special Acce

23、ssory消化爐特殊附件July 05/KJ33China_Kjeldahl SolutionsReflux module 回流模塊適用于必須在回流狀況下才COD-Accessory 化學(xué)耗氧量附件(僅適用于 K-437 and K-438)Determination of the Chemical Oxygen DemandDigestion Units: Special Accessory消化爐特殊附件July 05/KJ34China_Kjeldahl SolutionsCOD-Accessory 化學(xué)耗氧量附件DeterminaBchis Distillation Units 蒸餾單元

24、July 05/KJ35China_Kjeldahl SolutionsBchis Distillation Units 蒸餾單S T E A M最新的蒸餾單元 K-350 and K-355July 05/KJ36China_Kjeldahl SolutionsS T E A M最新的蒸餾單元 K-350 and K-老型號(hào) K-314Basic instrument for Kjeldahl and Nitrogen determinationDisadvantages K-314 no timer old technology price-performance ratioJuly 05

25、/KJ37China_Kjeldahl Solutions老型號(hào) K-314Basic instrument for蒸餾單元 K-350 and K-355總介紹Distillation Unit K-350K-314替代產(chǎn)品Distillation Unit K-355凱氏定氮的延伸產(chǎn)品July 05/KJ38China_Kjeldahl Solutions蒸餾單元 K-350 and K-355總介紹DistilK-350的特點(diǎn) 用于凱氏氮檢測(cè)的基本型號(hào) 定時(shí)器 可存儲(chǔ)一個(gè)方法: 蒸餾時(shí)間和試劑量 緊湊設(shè)計(jì) 安全特點(diǎn): 消化管在位傳感器, 安全門 以及維修門傳感器July 05/KJ39C

26、hina_Kjeldahl SolutionsK-350的特點(diǎn) 用于凱氏氮檢測(cè)的基本型號(hào)July 05/KK-355的特點(diǎn) 具有 K-350 全部功能，另加： 蒸汽輸出量可調(diào) (30 100 %) 額外配置抗強(qiáng)酸的泵 (用于加強(qiáng)酸和水) 冷凝水控制傳感器 可以儲(chǔ)存9個(gè)程序: 蒸餾時(shí)間, 試劑量 和蒸汽輸出量不僅適用于凱氏定氮，還適用于更廣泛的的應(yīng)用July 05/KJ40China_Kjeldahl SolutionsK-355的特點(diǎn) 具有 K-350 全部功能，另加：Jul 預(yù)算不高 樣品數(shù)量不多 K-350: 應(yīng)用于Kjeldahl and DevardaK-350/ K-355的目標(biāo)客戶

27、 K-355: 應(yīng)用于Kjeldahl, Devarda 和 非凱氏應(yīng)用在樣品準(zhǔn)備中需要蒸溜應(yīng)用的July 05/KJ41China_Kjeldahl Solutions 預(yù)算不高K-350/ K-355的目標(biāo)客戶 K-355: 我們的口號(hào) “STEAM”S安全操作T節(jié)約時(shí)間和成本E高效蒸餾A官方認(rèn)可的方法M 多用途設(shè)計(jì)July 05/KJ42China_Kjeldahl Solutions我們的口號(hào) “STEAM”S安全操作T節(jié)約時(shí)間和成本E蒸餾單元 B-324內(nèi)置滴定儀連接裝置 可以升級(jí)為全自動(dòng)的蒸餾單元電子蒸汽調(diào)節(jié)內(nèi)置泵系統(tǒng)用于NaOH, 水 和硼酸的添加消化管自動(dòng)排空存儲(chǔ)20種方法操作

28、簡(jiǎn)單, LCD 顯示July 05/KJ43China_Kjeldahl Solutions蒸餾單元 B-324內(nèi)置滴定儀連接裝置 July 05/KB-324的目標(biāo)客戶想要操作更舒適的想要連接滴定單元的客戶應(yīng)用于Kjeldahl, Devarda 和 steam distillation蒸汽蒸餾的客戶July 05/KJ44China_Kjeldahl SolutionsB-324的目標(biāo)客戶想要操作更舒適的July 05/KJ44蒸餾單元 K-370 蒸餾和滴定全部?jī)?nèi)置于一臺(tái)儀器 帶有彩色顯示屏的大控制面板 最新的軟件設(shè)計(jì)，帶有 - 方便的數(shù)據(jù)輸入輸出功能 - 多任務(wù)處理 - 多語(yǔ)言 - 高

29、存貯性能10000數(shù)據(jù) - 用戶可以自定義方法 緊密，功能型設(shè)計(jì) 配備 K-371 自動(dòng)進(jìn)樣器，可以實(shí)現(xiàn)自動(dòng)進(jìn)樣July 05/KJ45China_Kjeldahl Solutions蒸餾單元 K-370 蒸餾和滴定全部?jī)?nèi)置于一臺(tái)儀器July 稱量:自動(dòng)樣品量輸入精確的液體加樣器:用于反滴定自動(dòng)型消化爐 K-438:Control and Documentation of Digestion ProcessPC:數(shù)據(jù)轉(zhuǎn)移和備份自動(dòng)進(jìn)樣器:多樣品的自動(dòng)處理打印機(jī):外接 A4 Printer支持 PCL3蒸餾單元 K-370July 05/KJ46China_Kjeldahl Solutions稱

30、量:精確的液體加樣器:自動(dòng)型消化爐 K-438:PC:自動(dòng)自動(dòng)進(jìn)樣器 K-371 K-437 & K-438的 20 管架子可以 適合自動(dòng)進(jìn)樣器 特殊架子: 4 管 Express Rack樣品通過K-370控制 利用蒸汽實(shí)現(xiàn)樣品的轉(zhuǎn)移由樣品架到蒸餾單元July 05/KJ47China_Kjeldahl Solutions自動(dòng)進(jìn)樣器 K-371 K-437 & K-438的 20 自動(dòng)進(jìn)樣器 K-370/371自動(dòng)進(jìn)樣器K-371蒸餾儀K-370樣品轉(zhuǎn)移原理July 05/KJ48China_Kjeldahl Solutions自動(dòng)進(jìn)樣器 K-370/371自動(dòng)進(jìn)樣器K-371蒸餾儀K-自動(dòng)

31、進(jìn)樣器 K-370/371自動(dòng)進(jìn)樣器K-371蒸餾儀K-370蒸餾過程中蒸汽的走向July 05/KJ49China_Kjeldahl Solutions自動(dòng)進(jìn)樣器 K-370/371自動(dòng)進(jìn)樣器K-371蒸餾儀K-自動(dòng)進(jìn)樣系統(tǒng) K-370/371回收率 (NH4)H2PO4July 05/KJ50China_Kjeldahl Solutions自動(dòng)進(jìn)樣系統(tǒng) K-370/371回收率 (NH4)H2PO4Customers who want maximum comfortCustomers with high sample 通量 ( 15 樣品/天)需要標(biāo)準(zhǔn)化處理的客戶需要廣泛認(rèn)證的客戶K-37

32、0/371的目標(biāo)客戶July 05/KJ51China_Kjeldahl SolutionsCustomers who want maximum K-3Last but not least精確的接口制造用于和排廢裝置的緊密聯(lián)接，有效防止煙霧泄漏專業(yè)的冷凝裝置，避免飛濺和硫酸的過量消耗更寬的直徑 48mm= 對(duì)起沫樣品更有利300 ml的大容量 = 可以消化大量的樣品 48 mmGlassblowing one of Bchis Key Competences玻璃吹制Buchi公司核心競(jìng)爭(zhēng)力之一July 05/KJ52China_Kjeldahl SolutionsLast but not le

33、ast精確的接口制造用于和排Distillation Units: Special Accessory蒸餾單元的專業(yè)附件Devarda 防濺保護(hù)檢測(cè)NO2- 和NO3- 必需的附件 (例： 肥料) July 05/KJ53China_Kjeldahl SolutionsDistillation Units: Special AcTrouble Shooting July 05/KJ54China_Kjeldahl SolutionsTrouble Shooting July 05/KJ54C儀器校驗(yàn)方法 Distillation蒸餾標(biāo)準(zhǔn)物質(zhì)磷酸二氫氨(無(wú)水）(NH4)H2PO4 (water free)理論含量: 12.17 % Digestion and Distillation消化和蒸餾標(biāo)準(zhǔn)物質(zhì):甘氨酸Glycin (99.7%)理論含量: 18.66 %July 05/KJ55China_Kjeldahl Solutions儀器校驗(yàn)方法 Distillation蒸餾標(biāo)準(zhǔn)物質(zhì)磷酸二氫Trouble Shooting: 結(jié)果太低 Digestion消化過程 時(shí)間是否太短? 硫酸量是否太少? 消化溫度是否太低? 硫酸比率 : 消化是否混合?Distillation蒸餾N