Monitoring Chemical Changes in Cheddar Cheese During Aging by High Performance Liquid Chromatography and Gas Chromatography Techniques

The concentrations of several chemical metabolites in Cheddar cheese were monitored by various chromatographic techniques during the aging process to learn which metabolites were the best predictors of the glycolytic, lipolytic, and proteolytic age of the cheese. Pyruvic, lactic, acetic, and propionic acids were measured by ion-exchange high performance liquid chromatography; acetone, 2-butanone, ethanol, 2-pentanone, 2-butanol, and n-propanol were monitored by headspace gas chromatography; free fatty acids were quantitated (without derivatization) by gas chromatography; and free amino acids were determined as their o-phthaldehye derivatives by high performance liquid chromatography.The best predictors of the glycolytic age were propionic acid and acetic acid; the best predictors of lipolysis were the free fatty acids C₁₀, C₁₂, C₁₄, and C₁₆; and the best predictors of proteolysis were the free amino acids leucine, methionine, and glutamic acid. The volatile metabolites determined by headspace gas chromatography were not good indicators of aging; however, they did provide useful information related to flavor problems.Cheddar cheese aged at elevated temperatures produced propionic acid, acetic acid, and free amino acids at significantly faster rates than the other chemicals that were monitored.     

卤猪肉加工过程中游离脂肪酸、游离氨基酸及核苷酸变化分析

利用气相色谱质谱联用和高效液相色谱法检测分析了卤猪肉加工过程中游离脂肪酸、游离氨基酸及核苷酸的变化。结果表明,在加工过程中,总游离脂肪酸有效峰面积呈下降趋势,饱和脂肪酸也呈现下降的变化趋势,不饱和脂肪酸含量先增加后减少,在高温煮制1 h时达到最大值。总游离氨基酸的含量在卤猪肉加工过程中变化明显,在原料肉中总游离氨基酸含量为244.39 mg/100 g,其它样品中含量都显著(p0.05)高于原料肉,在二次煮制完成后达到最大值,为349.32 mg/100 g。谷氨酸在成品中的含量达到119.02mg/100 g,占总游离氨基酸的34.12%。呈味核苷酸检测结果显示,在加工过程中5'-IMP和5'-GMP的含量呈现下降趋势。由于加热导致核苷酸热降解,其降解产物肌苷的含量呈现上升趋势,在成品中检测到其含量显著(p0.05)高于原料肉中的含量。本研究结果为改进卤猪肉的生产工艺、提高产品品质提供了理论参考。     

Quantitative Determination of Lactic and Succinic Acids in Frozen Whole Eggs by Gas-Liquid Chromatography

SUMMARY— Frozen whole egg containing known amounts of lactic and succinic acids were evaluated using gas-liquid chromatography (GLC) and AOAC (1960) procedures. The acids were isolated from the whole egg samples by liquid-liquid extraction, evaporated to dryness, and refluxed for 2 hr with 1-butanol and HCI and chromatographed as their butyl ester derivatives, along with an internal standard. Peak heights and responses relative to butyl decanoate as an internal standard were used to calculate acid concentrations in the whole egg samples.
Lactic acid recovered from whole egg samples by GLC procedures ranged from 98% at the low concentration (0.43 mg/100 g egg) to 99% at the high concentration (47.70 mg/100 g egg). Succinic acid recovered from whole egg samples by GLC procedures ranged from 103% at the low concentration (1.22 mg/100 g egg) to 98% at the high concentration (73.20 mg/100 g egg). Acid recoveries from whole egg samples by GLC procedures were as accurate, and generally less variable, than those recovered by AOAC (1960) procedures.     

我国沿海主要海域雌性三疣梭子蟹呈味成分含量的比较

以我国渤海、黄海、东海和南海4个海域中雌性三疣梭子蟹的肝胰腺、性腺和体肉为研究对象,采用电子舌、氨基酸自动分析法以及高效液相色谱法研究不同海域的生长环境对雌性三疣梭子蟹3个可食部位游离氨基酸含量和呈味核苷酸化合物的变化,并采用滋味强度值和味精当量对呈味成分的强度进行评价。结果表明:电子舌可对4个海域中雌性三疣梭子蟹的肝胰腺、性腺和体肉进行有效区分;在肝胰腺部分,游离氨基酸总量是3个可食部位最高的,3种呈味核苷酸中肌苷酸和腺苷酸的含量较高;在不同海域中,渤海肝胰腺组游离氨基酸总量最高,为3 315.05 mg/100 g,且其味精当量值最高,为15.87 g MSG/100 g;南海肝胰腺组游离氨基酸总量最低,但甜味氨基酸含量所占比例最高,为63.10%,并且呈味核苷酸含量最高,肌苷酸和腺苷酸的滋味强度值大于1。蟹的性腺部分,呈味核苷酸含量与味精当量值是3个可食部位中最高的,其中南海性腺的味精当量值为36.46 g MSG/100 g,是4个海域中最高。蟹的体肉部分中肌苷酸含量较高,黄海体肉肌苷酸组中含量较低,其滋味强度值小于1,对体肉的滋味无直接贡献,但黄海体肉组游离氨基酸总量最高,为2 564.44 mg/100 g;东海体肉组的味精当量值最高,为7.19 g MSG/100 g。     

Beef, chicken and lamb fatty acid analysis - a simplified direct bimethylation procedure using freeze-dried material

When fractionation of meat lipids is not required, procedures such as saponification can be used to extract total fatty acids, reducing reliance on toxic organic compounds. However, saponification of muscle fatty acids is laborious, and requires extended heating times, and a second methylation step to convert the extracted fatty acids to fatty acid methyl esters prior to gas chromatography. Therefore the development of a more rapid direct methylation procedure would be of merit. The use of freeze-dried material for analysis is common and allows for greater homogenisation of the sample. The present study investigated the potential of using freeze-dried muscle samples and a direct bimethylation to analyse total fatty acids of meat (beef, chicken and lamb) in comparison with a saponification procedure followed by bimethylation. Both methods compared favourably for all major fatty acids measured. There was a minor difference in relation to the C18:1 trans 10 isomer with a greater (P<0.05) recovery with saponification. However, numerically the difference was small and likely as a result of approaching the limits of isomer identification by single column gas chromatography. Differences (P<0.001) between species were found for all fatty acids measured with no interaction effects. The described technique offers a simplified, quick and reliable alternative to saponification to analyse total fatty acids from muscle samples.     

美拉德反应对金枪鱼红肉酶解液挥发性物质和游离氨基酸的影响

为了高值化利用金枪鱼红肉这一蛋白质资源,利用气相色谱-质谱联用(GC-MS)和高效液相色谱(HPLC),研究美拉德反应对金枪鱼红肉酶解液挥发性物质和游离氨基酸的影响。GC-MS结果表明,金枪鱼红肉酶解液中共检测出19种挥发性化合物,其中未加热的酶解液中有10种,而经过不同时间(30、60和90 min)加热的酶解液中,分别检测出12种、15种和15种,其中呋喃类物质的含量随反应时间延长显著增加。HPLC分析表明,随着加热时间的增加,酶解液中呈味氨基酸总量均上升,但是鲜味和甜味氨基酸的比例呈下降趋势,苦味氨基酸则呈上升趋势。表明,美拉德反应赋予酶解液更加丰富的风味的同时,却使其呈现更多的苦味。     

高效液相色谱法检测食品中17种游离氨基酸

目的使用异硫氰酸苯酯衍生-高效液相色谱法对食品中17种游离氨基酸进行检测。方法对17种游离氨基酸及其苯胺硫甲酰基衍生物稳定性的分析,研究游离氨基酸的最佳衍生条件。同时,对盐沉淀法和C_(18)固相萃取(solid phase extraction, SPE)小柱法2种样品除杂方法的加标回收率进行测定。结果胱氨酸加热干燥5h后,回收率为68.46%±4.01%,热稳定性差;其它16种氨基酸加热1～5h后回收率均在90%～110%之间,热稳定性较好。胱氨酸的苯胺硫甲酰基衍生物,6h后回收率仅为11.04%±1.65%,稳定性差;其它氨基酸的苯胺硫甲酰基衍生物24 h后回收率均大于90%,比较稳定。另外,前处理中使用C_(18) SPE除杂时,样品中游离氨基酸的加标回收率均大于70%,而使用水杨酸和三氯乙酸除蛋白后苯胺硫甲酰基衍生物降解严重。结论除胱氨酸外,其余16种氨基酸的热稳定性均较好,且生成的苯胺硫甲酰基衍生物比较稳定,可以通过异硫氰酸苯酯衍生进行定量分析。相比于水杨酸和三氯乙酸沉淀法, C_(18) SPE柱是游离氨基酸除杂方法的首选,而无机盐影响异硫氰酸苯酯与氨基酸衍生,因此此方法不适用于含盐样品的衍生。     

不同宰杀方式对草鱼肉呈味水溶性成分的影响

以不同宰杀方式草鱼的背部肉为对象,采用高效液相色谱法和氨基酸自动分析法研究不同宰杀方式（自然、急杀、去鳃）对草鱼背部肉核苷酸类化合物和游离氨基酸含量的变化,并测定背部肉pH值、糖原和乳酸的含量。结果表明：自然、急杀、去鳃3 种宰杀方式的草鱼背肉苦味氨基酸含量占总游离氨基酸含量分别为78.95%、68.60%、69.44%,部肉的鲜度K值分别为20.28%、15.49%、12.13%,自然死亡组腺嘌呤核苷三磷酸含量最低,肌苷酸含量最低,鲜度最差。草鱼背肉中pH值的变化与乳酸含量的变化呈负相关,乳酸含量的变化与糖原含量的变化呈负相关,自然死亡的草鱼肉乳酸含量最高,pH值最低。不同宰杀方式对草鱼背肉呈味水溶性成分影响较大,自然死亡的草鱼肉滋味最差,急杀致死和去鳃致死组滋味差别不明显。因此,在加工过程中,要避免鱼血渗入肌肉,造成鱼肉土腥味产生。     

CHANGES IN NITROGEN FRACTIONS AND FREE AMINO ACIDS DURING RIPENING OF SPANISH DRIED BEEF "CECINA"

The changes in nitrogen fractions (total nonprotein nitrogen -NPN-, amino acid nitrogen, peptide nitrogen and volatile basic nitrogen) and free amino acids during the ripening of "cecina" (a Spanish dried beef product) were tracked. The amount of total NPN, amino acid nitrogen and volatile basic nitrogen increased during processing, while peptide nitrogen decreased in the last stage. An increase was observed in all amino acids except taurine. In the final product, the most abundant free amino acids were leucine-isoleucine mixture, lysine and proline-methionine mixture. Taking into account the concentrations of the most abundant amino acids detected in cecina and the taste thresholds described for L-amino acids in water, it appears that several amino acids i.e. lysine, alanine, valine and glutamic acid, could contribute to the flavor of this meat product.     

烟草中游离氨基酸的分析方法及研究方向

游离氨基酸是烟草中的重要化合物,对烟草香味物质的形成有着重要的作用。烟草游离氨基酸的分离和定量鉴定方法主要有：9－芴氯甲酸甲酯作为预柱衍生试剂的高压液相色谱法、氯甲酸乙酯作为衍生试剂的毛细管气相色谱法以及氨基酸自动分析仪法。其中,氨基酸自动分析仪更适合分离和鉴定烟草游离氨基酸。同时指出,对低焦油混合型卷烟－白肋烟进行氨基酸分析,以期找出最佳工艺,可以更有效地提高我国混合型卷烟的质量。     

蒸制鲟鱼肉特征性滋味组分的鉴定

为明确传统热加工鲟鱼肉特征性滋味组分,对蒸制鲟鱼肉中游离氨基酸、核苷酸、有机酸、甜菜碱及无机离子等滋味组分进行定量分析,并结合滋味活性值、滋味组分减缺、添加及重组实验探究蒸制鲟鱼肉的关键味觉化合物。结果表明：蒸制16 min鲟鱼肉的关键味觉化合物为甘氨酸、谷氨酸、丙氨酸、5’-一磷酸腺苷、5’-一磷酸肌苷、乳酸、琥珀酸、K＋、Na＋和Cl－;将10 种关键味觉化合物按其天然含量溶解在超纯水中制备简化重组液,简化重组液能够较为完整地复制完全重组液的味道,但与天然提取液相比,在鲜味上仍有所欠缺。     

Hydrolysis and Maillard Reactions During Ripening of Iberian Ham

Thirty-eight hams from Iberian breed were processed by the traditional dry-curing procedure. Active proteolysis and autooxidation occur during the first period, as revealed by the accumulation of peptide, nitrogen and carbonyl compounds. In the second step a strong increase in amino acid nitrogen was observed, whereas the content of all the aldehydes studied decreased notably. This could be due to condensations between carbonyls and free amino acids during the summer period. While hams were kept in the cellar, the overall increase of amino acid nitrogen was in contrast to the slow rate of accumulation of some individual amino acids. This was presumably due to degradation to volatile derivatives, where Maillard reactions could be involved.     

Pork meat quality affects peptide and amino acid profiles during the ageing process

Twenty pork carcasses were classified in different pork meat qualities: red, firm and non-exudative (RFN), pale, soft and exudative (PSE), red, soft and exudative (RSE) and dark, firm and dry (DFD) meat. The content of peptides and free amino acids during the ageing process was analysed and compared within quality classes. Four peptide fractions were isolated through cation-exchange and reverse-phase chromatography. The main significant differences among qualities were obtained for peptide fractions 3 and 4. Peptide fraction 3 at 4 days and peptide fraction 4 at 2 h postmortem were higher in the ideal pork quality (RFN) than in the other quality classes. The ageing of pork meats produced a general increase in all free amino acid concentrations for the studied quality classes except for Gln, β-Ala, Taurine and Orn and the dipeptides carnosine and anserine. The DFD class showed higher increases in Lys, Ala and Met probably due to the activation of neutral aminopeptidases.     

火腿风味调味料加工过程中脂解的变化研究

以复合蛋白酶和中性脂肪酶为发酵剂在微控条件下生产火腿风味调味料,研究了此生产过程中脂解的变化。通过比较不同时期样品的气相色谱结果发现:从鲜肉到酶解后游离脂肪酸均以棕榈酸,油酸,硬脂酸和亚油酸为主,且含量不断积累;整个加工过程中,脂肪酸的释放速率为硬脂酸>棕榈酸>亚油酸>油酸;后期的加工过程中,油酸和亚油酸发生氧化降解,分别从水解后的1.638%、0.8276%,下降到氧化后的1.218%、0.5975%;而饱和脂肪酸含量几乎不变。     

冻结方式对不同部位草鱼呈味物质的影响

以草鱼腹肉、背肉和红肉为研究对象,采用－40?℃速冻、－20?℃乙醇液体冻结和－20?℃静止空气冻结对草鱼进行冻结处理,采用高效液相色谱法、氨基酸自动分析法以及电子舌测定不同冻结方式的鱼肉中呈味物质含量变化,并测定各部位鱼肉的乳酸含量。结果表明,冻结方式对鱼肉的鲜度和呈味物质含量有一定影响。肌苷酸（inosine monphosphate,IMP）和一磷酸腺苷（adenosine monophosphate,AMP）含量、游离氨基酸总量与冻结速率呈正相关,但乳酸、次黄嘌呤核苷（insoine,HxR）和次黄嘌呤（hypoxanthine,Hx）的含量与冻结速率呈负相关,在－20?℃冻藏条件下贮藏3?d后,－40?℃速冻组K值最小,－20?℃静止空气冻结组K值最大。红肉部分IMP、AMP含量低,而HxR和Hx含量较高,其滋味和鲜度较差,3?种冻结方式中红肉的K值分别为28.99%、40.92%、46.58%;腹肉与背肉中游离氨基酸含量相近,且高于红肉部位,但红肉中天冬氨酸、谷氨酸含量明显高于腹肉和背肉;红肉中乳酸含量显著高于腹背肉,且不同冻结方式对其影响较大。电子舌能有效区分不同冻结方式和不同部位的草鱼肉。     

Concentration of Umami Compounds in Pork Meat and Cooking Juice with Different Cooking Times and Temperatures

This study examined the concentrations of umami compounds in pork loins cooked at 3 different temperatures and 3 different lengths of cooking times. The pork loins were cooked with the sous vide technique. The free amino acids (FAAs), glutamic acid and aspartic acid; the 5′‐nucleotides, inosine‐5′‐monophosphate (IMP) and adenosine‐5′‐monophosphate (AMP); and corresponding nucleoside inosine of the cooked meat and its released juice were determined by high‐performance liquid chromatography. Under the experimental conditions used, the cooking temperature played a more important role than the cooking time in the concentration of the analyzed compounds. The amino acid concentrations in the meat did not remain constant under these experimental conditions. The most notable effect observed was that of the cooking temperature and the higher amino acid concentrations in the released juice of meat cooked at 80 °C compared with 60 and 70 °C. This is most likely due to the heat induced hydrolysis of proteins and peptides releasing water soluble FAAs from the meat into the cooking juice. In this experiment, the cooking time and temperature had no influence on the IMP concentrations observed. However, the AMP concentrations increased with the increasing temperature and time. This suggests that the choice of time and temperature in sous vide cooking affects the nucleotide concentration of pork meat. The Sous vide technique proved to be a good technique to preserve the cooking juice and the results presented here show that cooking juice is rich in umami compounds, which can be used to provide a savory or brothy taste.     

火腿风味料加工过程中游离脂肪酸的变化

以复合蛋白酶和复合脂肪酶为发酵剂。在微控条件下生产火腿风味调味料,研究了此生产过程中游离脂肪酸的变化。通过比较不同时期样品的色谱结果发现:从鲜肉到酶解后游离脂肪酸均以棕榈酸,油酸,硬脂酸和亚油酸为主。且含量不断积累;整个加工过程中,脂肪酸的释放速率为硬脂酸>棕榈酸>亚油酸>油酸;后期的加工过程中,油酸和亚油酸发生氧化降解,分别从水解后的1.638%、0.8276%.下降到氧化后的1.218%、0.597 5%;而饱和脂肪破含量几乎不变。     

不同加工方式下不同部位梅花鹿肉中氨基酸的含量分析

为研究不同加工方式下不同部位梅花鹿肉中氨基酸的含量差异及变化趋势,本研究采用阳离子交换色谱法分别测定了冻干及烘干8个不同部位鹿肉的17种水解氨基酸含量;并对两种不同加工方式下总氨基酸、必需氨基酸和呈味氨基酸含量进行了比较分析.结果表明,烘干梅花鹿肉总氨基酸含量为55.36～73.66 g/100 g;冻干梅花鹿肉总氨基...     

Free amino acids by high performance liquid chromatography and peptides by gel electrophoresis in Mahon cheese during ripening

The free amino acids and peptides contents of Mahon cheese during four months of ripening were studied using high performance liquid chromatography and gel electrophoresis, respectively. The total content of free amino acids generally increased throughout the ripening period, except in one batch. Phenylalanine, valine, proline, glutamic acid and isoleucine were the most abundant free amino acids in all the tests throughout the four month period. They accounted for between 67 and 80% of the free amino acids. No difference was found between the pattern of traditionally made and industrially made Mahon cheeses. Electrophoresis of soluble nitrogen showed the main whey proteins as well as other bands corresponding to peptides.     

Identification of candidate amino acids involved in the formation of pink-red pigments in onion (Allium cepa L.) juice and separation by HPLC

The formation of pink-red pigments ("pinking") by various amino acids was investigated by reacting amino acids with compounds present in onion juice. The unknown pink-red pigments were generated and separated using high-performance liquid chromatography (HPLC) and a diode array detector (DAD) in the range of 200 to 700 nm. To generate pink-red pigments, we developed several reaction systems using garlic alliinase, purified 1-propenyl-L-cysteine sulfoxide (1-PeCSO), onion thiosulfinate, natural onion juice, and 21 free amino acids. The compound 1-PeCSO was a key compound associated with pinking in the presence of both the alliinase and amino acids. Numerous naturally occurring pink-red pigments were detected and separated from pink onion juice using the HPLC-DAD system at 515 nm. Most free amino acids, with the exceptions of histidine, serine, and cysteine, formed various pink-red pigments when reacted with onion thiosulfinate. This observation indicated that onion pinking is caused not by a single pigment, but by many. Furthermore, more than one color compound could be produced from a single amino acid; this explains, in part, why there were many pink-red compound peaks in the chromatogram of discolored natural onion juice. We presumed that the complexity of the pink-red pigments was due to the involvement of more than 21 natural amino acids as well as several derivatives of the color products produced from each amino acid. We observed that the pinking process in onion juice is very similar to that of the greening process in crushed garlic, emphasizing that both thiosulfinate from flavor precursors and free amino acids are absolutely required for the discoloration.