HPLC/UV determination of organic acids in fruit juices and nectars

A reversed phase HPLC method for separation and determination of organic acids in fruit juices and nectars is presented. The method is based on the reaction of free organic acids with O-(4 nitrobenzyl)-N,N'-diisopropylisourea (PNBDI) in presence of dioxane. Excess of reagent was removed with a strong cation-exchange resin. The p-nitrobenzyl esters were separated on a C₁₈ reversed phase column using gradient elution with water and acetonitrile and UV detection at 265 nm. Benzylmalonic acid was used as internal standard. The determinations were performed in the linear range of 0.05-2 g/l for lactic, acetic and succinic acids, 0.1-12 g/l for tartaric, malic and citric acids. The detection limits were 0.025 g/l, 0.017 g/l, 0.050 g/l, 0.039 g/l, 0.025 g/l and 0.060 g/l, respectively for lactic, acetic, tartaric, malic, succinic and citric acids. Validation of the method was carried out by the standard additions method, the recoveries ranged from 91.4 to 103.0%. The precision of the method was also evaluated and reported a CV% as less than 3.1%. Organic acid profiles of citrus fruit, pineapple and apple natural juices (home-made), commercial juices and nectars were established.     

啤酒中有机酸含量的RP-HPLC检测条件优化及其酿造过程中动态分析

该研究对反相高效液相色谱（RP-HPLC）法测定啤酒中有机酸含量的方法进行优化,最终确定了RP-HPLC法测定啤酒中草酸、乳酸、酒石酸、乙酸、苹果酸、α-酮戊二酸、抗坏血酸、柠檬酸、琥珀酸9种有机酸的检测条件,并结合发酵机理对啤酒酿造过程中的有机酸含量动态变化进行分析。结果表明,RP-HPLC优化条件为：检测波长215 nm、流动相缓冲液0.10 mol/L KH2PO4、pH值3.0、流速0.6 mL/min。有机酸在质量浓度0.2～400.0 mg/L范围内线性关系良好（R2均＞0.99）,加标回收率79.3%～110.0%,精密度试验结果相对标准偏差（RSD）为0.3%～11.5%,表明该方法精密度高、准确性良好。在啤酒酿造过程中有机酸总量及乙酸、琥珀酸、苹果酸、乳酸、柠檬酸、α-酮戊二酸含量呈现先增长后平稳的趋势,草酸、抗坏血酸、酒石酸含量变化相对平稳。     

桑椹酒发酵过程中有机酸含量变化

利用高效液相色谱法测定桑椹酒发酵过程中七种有机酸的变化,结果显示:桑椹原果汁中琥珀酸含量最高,柠檬酸次之;发酵过程中酒石酸、苹果酸、柠檬酸、琥珀酸、草酸、抗坏血酸含量变化均先呈上升趋势,达到最大值后呈下降趋势,而乳酸含量则一直上升;发酵结束后检测到酒石酸、苹果酸、柠檬酸、琥珀酸、草酸、抗坏血酸、乳酸含量分别为1.822、2.54、2.44、0.76、0.772、0.0756、0.0846g/L。     

葡萄酒酿造过程中有机酸变化规律研究

采用离子色谱法分析了葡萄酒中9种有机酸含量。通过对赤霞珠葡萄酒发酵过程中主要有机酸变化分析,研究其从葡萄汁到原酒的酿造过程中有机酸含量变化规律和量变的幅度。结果表明,葡萄酒中有机酸主要有酒石酸、苹果酸、乳酸、柠檬酸和琥珀酸,葡萄汁经发酵产生了乳酸、乙酸和琥珀酸。酿造过程中酒石酸、苹果酸、抗坏血酸和柠檬酸含量一直呈下降趋势;草酸和琥珀酸含量先增高后减少,乳酸含量一直呈上升趋势。乙酸是葡萄酒酒精发酵的主要副产物,含量为0.2～0.3 g/L。     

Simple determination of main organic acids in grape juice and wine by using capillary zone electrophoresis with direct UV detection

An accurate, simple and rapid capillary zone electrophoresis (CZE) method with direct UV detection has been set up for the determination of main organic acids in grape juice and wine. The determination of tartaric, malic, and citric acids in grape juices and tartaric, malic, succinic, acetic, lactic and citric acids in wines can be achieved in less than 3 min with only a simple dilution and filtration treatment of the sample. Validation parameters of the method as detection and quantification limits, linearity, precision (intraday and interday analysis) and recovery were also studied in grape juice, white wine, rose wine and red wine, separately. The proposed method decreases the analysis times of the previous reported CZE methods and allows the rapid control of the grape maturity, the winemaking processes and the detection of wine alterations and/or illnesses.     

利用反相高效液相色谱法测定梨汁中有机酸的种类和含量

研究了利用HPLC法分离测定梨汁中有机酸的条件 ,同时对来源于不同产地和品种梨汁中的有机酸的种类和含量进行了测定 ,并应用SAS统计软件对各有机酸含量之间的相关性进行了分析。结果表明 ,选择 0 0 1mol/LK2 HPO4 磷酸盐缓冲液 (pH 2 5 5 )和体积分数 3 %甲醇做流动相 ,流动相流速为 0 5mL/min ,柱温为 3 0℃ ,紫外检测波长为 2 1 0nm时 ,可以较好地分离和测定果汁中常见的 1 1种有机酸 ,该方法相对标准偏差 0 72 %～ 2 99% ,回收率 91 1 %～ 1 0 6 5 % ,各种酸的线性相关系数r>0 9996,具有较高的准确度和精确度 ;梨汁中的主要有机酸有苹果酸、柠檬酸、琥珀酸、莽草酸、酒石酸、奎宁酸、乳酸和富马酸 ,其中苹果酸含量最高 (为 1 1 4× 1 0 3～ 3 0 9× 1 0 3mg/L) ;不同品种梨汁中各有机酸含量差别较大 ,其中琥珀酸与乳酸和富马酸 ,莽草酸与柠檬酸和奎宁酸的含量之间表现出极显著相关性。     

原汁苹果醋中的有机酸

以苹果汁为原料 ,采用液态酒精发酵和醋酸发酵法制作苹果醋 ,采用反相高效液相色谱法分析苹果醋中有机酸 ,研究了苹果品种、醋酸菌种、醋酸发酵方法及过程对有机酸的影响。认为苹果酸是苹果醋中的主要有机果酸 ,其与液态粮食醋有机酸组成的差别主要体现在苹果酸、酒石酸和乳酸的含量上 ,苹果品种是影响原汁苹果醋中有机酸的种类和含量主要因素 ,醋酸菌在醋酸发酵过程中有代谢各种有机酸的作用 ,发酵方法会影响醋酸菌对有机酸代谢作用的程度 ,试验中所采用的 5种醋酸菌在醋酸发酵过程中通过消耗苹果酸、乳酸、琥珀酸 ,而产生和积累柠檬酸。     

Effects of Product Formulation, Processing, and Meal Composition on In Vitro Estimated Iron Availability from Cereal-Containing Breakfast Meals

The effects of breakfast cereal composition, breakfast cereal processing, and breakfast meal composition on relative iron availability from breakfast meals were estimated using an in vitro method. Addition of wheat bran and germ to cereals and meals reduced iron availability. Presweetening caused a slight increase in iron availability. Comparisons among meals containing cereals that were similar except for type of processing showed that processing may affect iron availability. Addition of orange juice to breakfast meals caused a dramatic enhancement of iron availability. Ascorbic acid fortified apple and grape juice increased iron availability but the effect was small compared with the orange juice effect. Studies with purified organic acids showed that ascrobic and citric acids present in the juices caused the observed enhancement of iron availability. Citric acid was a more potent enhancer than ascorbic acid.     

Validation of a HPLC method for simultaneous determination of main organic acids in fruits and juices

A liquid chromatographic method for fast and simultaneous determination of tartaric, malic, ascorbic and citric acids was validated for further application to fruits and juices. Moreover, the organic acids content of commercial samples of fruits and juices were evaluated, as well as the ascorbic acid stability during the storage. Determination of organic acids was carried out using a liquid chromatograph coupled to a diode array detector, with reversed phase (C₁₈ column) and isocratic elution with 0.01 mol L⁻¹ KH₂PO₄ (pH = 2.60) mobile phase. The validation parameters showed efficiency, adequate linearity, relative standard deviation values between 0.4% and 2.3% (n = 10) for repeatability and from 1.2% to 5.0% (n = 18) for reproducibility, limits of detection (LD) were between 0.03 and 3.31 μg mL⁻¹ and quantification (LQ) were between 0.10 and 11.03 μg mL⁻¹, recovery rates were between 82% and 110%, for two levels. In addition, the method is fast (10 min) and generates low and non-toxic residues. The values found for vitamin C were about 10 times above the values declared at the package. Ready to drink juices have a composition similar to the fruit, concerning to organic acids, except for the powder juice, in which only ascorbic and citric acids were found, for all tastes. After opening the package, a decrease of 14.0% and 27.0% in ascorbic acid content was observed for orange powder and ready to drink juices, respectively.     

Determination of organic acids in fruits and vegetables by liquid chromatography with tandem-mass spectrometry

A method using liquid chromatography coupled to tandem-mass spectrometry (LC-MS/MS) with triple quadrupole in selective reaction monitoring mode was developed to determine organic acids (glutamic, tartaric, quinic, malonic, malic, shikimic, α-ketoglutaric, pyruvic, citric, succinic and fumaric acids) in fruits (melon, grape, peach, orange, lemon) and vegetables (green and red pepper, tomato, lettuce and lamb’s lettuce). The negative ion mode of ESI was used. All the validation parameters were found to be within the range of acceptance for all the compounds. Different matrix effects were observed in the studied vegetables and samples were quantified by the standard additions approach. The main advantages of this method are its selectivity, which is one of the main weaknesses of most methods when analysing organic acid in complex matrices, and its simplicity since little sample preparation is needed.     

HPLC双检测器法同时测定饮料中的有机酸与糖

本文提出了用高效液相色谱法同时测定饮料及果汁中的酸味物质与甜味物质。用交联度为8％的氢型阳离子交换树脂Aminex HPX-87H色谱柱,对各种有机酸及糖分的保留性质进行了研究。以0.01NH_2SO_4为移动相,糖分及有机酸分别得到了较好分离。利用糖分及有机酸对紫外光吸收强弱的差异,采用双检测器系统,使有机酸及糖分均能在对其具有较高灵敏度的检测器中得到检测。因而一次直接进样可同时定性及定量分析待测样品中的柠檬酸、酒石酸、苹果酸 (或维生素C) 琥珀酸、蔗糖、葡萄糖及果糖。每一样品的分析过程不超过十五分钟,这是其它方法很难达到的,特别对样品数量较多的日常分析,本方法的优越性更为明显。     

发酵前后甜石榴汁中有机酸的变化研究

本试验利用反相液相色谱（RP-HPLC）法分离测定发酵前后甜石榴汁中有机酸种类和含量。在甜石榴汁中检测到的有机酸包括草酸、苹果酸、α-酮戊二酸、乳酸、乙酸、柠檬酸与富马酸,其中主要有机酸为乳酸（占总有机酸的51.2%）、草酸（占总有机酸的26.8%）和柠檬酸（占总有机酸的16.8%）。发酵后甜石榴汁中的草酸与乳酸含量下降,乙酸含量增加,新生成酒石酸、丙酮酸与琥珀酸;苹果酸与柠檬酸含量增加。发酵后甜石榴汁的有机酸总量下降了7.8%.     

ACID TOLERANCE OF ESCHERICHIA COLI FOLLOWING COLD SHOCK TREATMENT

The effect of an initial cold shock treatment (2 h at 10C), following an abrupt downshift in temperature from 37 to 10C, on the subsequent growth and survival of Escherichia coli strains O157:H7 and MY20 (Biotype 1) in acidified Trypticase soy broth (TSB) and fruit juices (orange, apple) was investigated. Overall, no difference in growth at 37C was observed between each cold shocked and noncold shocked E. coli strain when cultured in TSB adjusted with either acetic acid (pH 6.0)or malic, citric and tartaric acid (each adjusted to: pH 4.5, 5.0, 5.5, 6.0). However, significant (P ± 0.05) differences in survival were observed between cold shocked and noncold shocked populations in TSB acidified with acetic acid (pH 5.0) or citric, malic and tartaric acid (pH 4.0). For strain MY20, survivor levels for cold shocked cells in TSB acidified with acetic acid citric, malic and tartaric acid at 8C were significantly (P ± 0.05) higher than in noncold shocked populations. Also, at 37C survival levels for cold shocked cells were significantly (P ± 0.05) higher than noncold shocked cells in TSB acidified with either malic or tartaric acid (pH 4.0). For the O157:H7 strain, survivor levels were higher (P ± 0.05) for cold shocked cells when maintained in TSB at 37C regardless of acid type. At 8C, cold shock treatment only increased (P ± 0.05) the survival of the O157:H7 strain in TSB adjusted with acetic acid (pH 6.0). Acid cross protection induced by cold shocking, as evidenced by enhanced survival, was not apparent for either E. coil strain in apple (pH 3.5) or orange juice (pH 3.8) maintained at 8C.     

未除气与除气淡爽型啤酒中9种有机酸的阈值

采用ASTM679标准方法测定了未除气与除气淡爽型啤酒中乙酸、乳酸、柠檬酸、苹果酸、琥珀酸、富马酸、丙酮酸、酒石酸和草酸的个人和小组阈值。结果发现,在未除气啤酒中,9种有机酸的小组阈值大小顺序为苹果酸、富马酸、乳酸、柠檬酸、丙酮酸、琥珀酸、乙酸、草酸、酒石酸。其中品评员对苹果酸、琥珀酸、富马酸、草酸和柠檬酸的口感差异相对较小,而对酒石酸、乙酸、丙酮酸和乳酸的口感差异较大。在除气啤酒中,9种有机酸的小组阈值大小顺序为丙酮酸、乳酸、乙酸、富马酸、琥珀酸、苹果酸、酒石酸、草酸、柠檬酸。品评员对柠檬酸、苹果酸和乳酸的口感差异相对较小,而对丙酮酸、草酸、酒石酸、富马酸、琥珀酸和乙酸的口感差异较大。此外还发现未除气和除气啤酒中有机酸阈值差异很大,其中差异最大的是三元酸柠檬酸,其次为二元酸,包括苹果酸、草酸、酒石酸、琥珀酸和富马酸,差异最小的是一元酸,包括乳酸、乙酸和丙酮酸。     

不同碳源对酵母代谢有机酸的影响

对啤酒酵母在以葡萄糖、果糖、麦芽糖作碳源的培养基中的代谢生长状况进行了跟踪检测,应用高效液相色谱检测各发酵液中酒石酸、苹果酸、琥珀酸和柠檬酸的含量变化,得出了碳源对啤酒酵母有机酸代谢的影响:其中酒石酸、苹果酸和柠檬酸含量差别明显,琥珀酸含量差别较小。     

Organic acids in orange,grapefruit and cherry juices quantified by high-performance liquid chromatography using neutral resin or propylamine columns

Malic and citric acids were measured in orange and grapefruit juices by h.p.l.c. using a neutral resin column. Low levels of succinic acid (<0.01%) present in some samples required the use of a propylamine column, which was also used for succinic and malic acids in sweet cherry juice. All samples required preliminary separation on ion exchange columns to remove interfering substances. Either refractive index or ultraviolet (206 nm) detectors can be used for analyses after purification.     

乙醇及有机酸对木醋杆菌合成细菌纤维素的影响

研究了乙醇、草酸、酒石酸、丙酮酸、苹果酸、乳酸、乙酸、柠檬酸、丁二酸对木醋杆菌产细菌纤维素的影响。结果发现:丙酮酸、乳酸、乙酸、乙醇、苹果酸、柠檬酸、丁二酸对木醋杆菌合成细菌纤维素有促进作用;草酸、酒石酸对木醋杆菌合成细菌纤维素有抑制作用;乙醇及各有机酸的最佳添加浓度分别为:丙酮酸0.15%,苹果酸0.1%,乳酸0.3%,乙酸0.4%,柠檬酸0.1%,丁二酸0.2%,乙醇4%;由正交实验得混酸的最适添加配比为:丙酮酸0.1%,苹果酸0.12%,乳酸0.2%,乙酸0.3%,柠檬酸0.12%,丁二酸0.2%,乙醇3%,BC产量达3.012g/L,是空白实验组的2.03倍。      

不同地区不同成熟度青梅有机酸的测定及相关性分析

目的：对四川达州青梅有机酸含量进行测定并分析其相关性,为产品深加工提供基础数据。方法：采用高效液相色谱法测定成熟度不同的青梅中有机酸含量,并对草酸、酒石酸、苹果酸、抗坏血酸、L-乳酸、乙酸、柠檬酸、丁二酸、齐墩果酸、熊果酸进行相关性分析。结果：同一地区,景市镇全熟黄果中柠檬酸、苹果酸均高于七成熟绿果,百节镇和渡市镇七成熟绿果中的柠檬酸均高于全熟黄果,而苹果酸反之。相关性分析表明,柠檬酸与熊果酸、L-乳酸、乙酸、齐墩果酸存在显著变化,与苹果酸无显著变化。结论：四川达州不同地区青梅中有机酸含量最高的是柠檬酸（19.546%）,其次是苹果酸（4.013%）,较其他产区青梅的高。     

草莓汁中糖酸组成分析及其在掺假鉴别中的应用

为了解草莓汁的糖酸组成特点并探索其在掺假鉴别中的应用,采用高效液相色谱法对10个品种草莓果汁中可溶性糖、有机酸组成与含量进行分析,结合化学计量学方法探索其在草莓汁掺假鉴别中的应用。结果表明,草莓汁中可溶性糖主要由蔗糖、葡萄糖和果糖组成,有机酸主要是柠檬酸和苹果酸。不同品种草莓果汁中各种可溶性糖和有机酸的含量存在较大差异,但葡萄糖与果糖的比值受品种影响较小,柠檬酸占检测到总有机酸含量的62.39%~82.73%。通过分析草莓原汁及其掺假样品中可溶性糖和有机酸组成,结合主成分分析和线性辨别分析等模式识别方法,可以实现对草莓果汁与掺加梨汁、苹果汁、杏汁和桃汁等掺假样品的区分,而且掺假量越大,区分效果越好。草莓果汁可溶性糖和有机酸组成特征结合化学计量学分析能够对掺假草莓果汁进行较好地区分,可用于草莓果汁掺假鉴定和质量控制。     

NONVOLATILE ORGANIC ACIDS IN GUAVA

ABSTRACT— The nonvolatile organic acids of guava were extracted and isolated. TLC of the acids showed the presence of 6 acids, 5 of the acids identified as lactic, malic, citric, ascorbic and galacturonic. GLC of the methyl esters of the guava acids confirmed the presence of malic, citric and lactic acid. Quantitative determinations using succinic acid as an internal standard showed citric and malic acid to be present in almost equal amounts and lactic acid in much lesser amount in cultivated guavas. In wild guavas, citric acid was the predominant acid, with lesser amounts of malic and lactic acids.