工业合成甘氨酸其含量的测定——甲醛法

<正> 甘氨酸是医药、有机合成和生物化学研究中常用的氨基酸之一。工业上,甘氨酸可由一氯醋酸和氢氧化铵作用而成。而工业合成甘氨酸其含量的测定目前尚无国家标准。上海市卫生局药品检验标准中,甘氨酸的含量测定,采用高氯酸—冰乙酸非水滴定法。其它氨基酸的含量测定在工业上和医药上亦主要采用非水滴定和克氏定氮法,但是工业合成甘氨酸,由于其样品中有氯化铵的存在,用以上两法测定其准确性均受干扰。本文介绍用过量的氧化镁去除NH_3,在     

小鼠脑内甘氨酸含量在缺氧预适应中的变化

在小鼠重复缺氧预适应过程中,用HPLC方法,测定其全脑与不同脑区中的甘氨酸含量。结果表明,随着动物对低氧耐受性的增高,其全脑、间脑,特别是海马、脑干中的甘氨酸含量升高。结果提示,甘氨酸作为抑制性递质对低氧预适应的形成具有正面影响。     

商品鹿鞭中氨基酸含量比较

采用氨基酸自动分析仪测定不同品种与产地的9份鹿鞭药材中氨基酸成分的种类及含量。这9份样品中均含17种氨基酸,其中人体必需氨基酸6种。17种氨基酸中以甘氨酸含量最高,必需氨基酸中亮氨酸含量最高;比较不同样品的含量,以辽宁西丰的梅花鹿鞭的总氨基酸含量最高,黑龙江梅花鹿鞭的必需氨基酸含量最高。结论为不同产地鹿鞭中氨基酸含量有一定差异,该方法可用于鹿鞭药材的质量评价。     

高效液相色谱法测定葡萄叶中甘氨酸甜菜碱、脯氨酸及葫芦巴碱

甘氨酸甜菜碱、脯氨酸及葫芦巴碱的含量与葡萄的抗逆性能密切相关,应用离子交换树脂提取葡萄叶中的这三种物质,采用Suger-pak1(waters)色谱柱,柱温:85℃;样品测定时间:70min;进样体积:100μL;流速:0.6mL/min;检测波长:195nm;流动相:50mg/LCa-EDTA的HPLC同时分析测定这三种物质,甘氨酸甜菜碱、脯氨酸及葫芦巴碱线性相关系数分别为0.9999,0.9994及0.9994;保留时间分别为20.28,25.66及30.65min;检出限分别为1.35,2.56及1.02ng/mL,回收率为93%~96%。此方法分离效果好,稳定,精确。     

HPLC测定不同海拔小叶金露梅17种氨基酸含量

目的:建立高效液相色谱法测定小叶金露梅中17种氨基酸含量的方法。方法:采用N,N-二硝基氟苯柱前衍生氨基酸后测定;色谱柱为Phenomenex Gemini 5μC18,流动相为醋酸钠(pH=6.40)和50%乙腈,采用梯度洗脱,检测波长为360nm,外标法计算含量。结果:17种氨基酸线性关系、精密度、稳定性、重复性良好,加样回收率为97.4%～102.8%(RSD为1.21%～2.5%)。各海拔的氨基酸总量范围是10%～14.79%,在不同海拔小叶金露梅中氨基酸平均质量分数顺序为亮氨酸>天冬氨酸>异亮氨酸>组氨酸>丝氨酸>精氨酸>丙氨酸>苯丙氨酸>缬氨酸>谷氨酸>脯氨酸>甘氨酸>赖氨酸。小叶金露梅含有7种必需氨基酸,它们的含量均大于48%。本方法简便、准确,为小叶金露梅药理提供理论依据。     

皱肋文蛤氨基酸含量及脂肪酸组成分析

测定了皱肋文蛤(Meretrix lyrata)软体部分的氨基酸含量与脂肪酸组成.共检出17种氨基酸,总含量为软体部干重的52.26%;4种呈味氨基酸(天门冬氨酸、谷氨酸、甘氨酸和丙氨酸)的含量为22.07%,占氨基酸总量的42.23%;必需氨基酸(EAA)总含量为20.72%,其必需氨基酸的构成比例基本符合FAO/W...     

甘氨酸和谷氨酸与钼配施对生菜品质的影响

以美国大速不结球生菜为材料,采用水培法研究了单一甘氨酸和谷氨酸分别替代营养液中20%硝态氮处理以及配合施加钼(0.002 mmol·L-1)处理对植株叶片鲜重和营养品质的影响.结果显示:(1)单一甘氨酸、谷氨酸替代处理培养12～16d,均可显著提高生菜叶片的鲜重和品质,其中甘氨酸对降低叶片硝酸盐含量、提高游离氨基酸和维生素C含量的效果好于谷氨酸,而谷氨酸促进生长和提高可溶性蛋白质、可溶性糖和叶绿素含量的效果好于甘氨酸.(2)甘氨酸与钼配合施用培养8～16 d的叶片鲜重显著高于单独甘氨酸替代处理,而谷氨酸与钼配合施用处理叶片鲜重与单独谷氨酸替代处理无显著差异;钼的配合施用进一步提高了甘氨酸、谷氨酸改善品质的作用效果,其叶片游离氨基酸、可溶性蛋白质、维生素C、可溶性糖和叶绿素含量显著提高,而硝酸盐含量显著降低,但甘氨酸与钼配合施用效果较好.研究表明,甘氨酸、谷氨酸均可作为生菜生长的氮源,并与钼配合施用对提高生菜产量和品质具有协同配合效应,且以甘氨酸与钼配施12 d效果较好.     

脑出血和脑梗死患者血清谷氨酸,甘氨酸和丙氨酸浓度的改变及意义

目的探讨血清中谷氨酸、甘氨酸和丙氨酸含量变化与脑出血和脑梗死的关系,为今后临床深入研究提供参考资料。方法用氨基酸分析仪对２３例急性脑出血和２６例脑梗死病人谷氨酸、甘氨酸和丙氨酸进行测定,并与对照组进行比较。结果病人组的兴奋性氨基酸谷氨酸的含量明显高于对照组（Ｐ＜０．０１）,病人组的抑制性氨基酸甘氨酸和丙氨酸的含量与对照组无显著差异（Ｐ＞０．０５）。结论提示兴奋性氨基酸谷氨酸可能作为损伤因素参与脑出血和脑梗死脑损伤的发生。     

丹参提取物中酚酸类含量的快速检测法

建立丹参提取物中酚酸类含量的快速检测方法。以丹酚酸B为对照,采用分光光度法于482 nm处测定丹参提取物中酚酸类的含量。结果表明丹酚酸B在0.0782～1.0166 mg(r=0.9977)范围内线性关系良好,回收率为101.652%,RSD值为0.81%(n=9)。该提取物中酚酸类含量测定结果为19.636%。该方法快速简便、准确、重现性好,可作为快速检测丹参中总酚酸含量的方法。     

高效亲和色谱法测定牛初乳加钙咀嚼片中免疫球蛋白IgG的含量

目的:建立高效亲和色谱法(HPAC)测定牛初乳加钙咀嚼片中免疫球蛋白IgG含量的方法。方法:采用HI-Trap Protein G HP亲和色谱柱(1 mL)色谱柱,以0.05 mol/L磷酸盐缓冲液和0.05 mol/L的甘氨酸-盐酸缓冲液为流动相,流速0.4 mL·min-1,检测波长280 nm,柱温30℃。结果:IgG含量在0.1502~0.7512 mg/mL浓度范围内与峰面积呈良好的线性关系(r=0.999 9);平均回收率为98.5%,RSD为1.2%(n=6)。结论:方法简便、快速,重现性好。可作为牛初乳加钙咀嚼片的质量控制方法。     

Sex differences in the bile acid composition of human bile: Studies in patients with and without gallstones

The bile acid composition of human gallbladder bile was studied in 83 subjects, 20 of each sex without discernible hepatobiliary disease, and 20 men and 23 women with cholelithiasis. The bile acids were measured by combined thin-layer and gas-liquid chromatography.In the bile of patients without cholelithiasis the molar percent of cholic acid was significantly greater in men while that of chenodeoxycholic acid was significantly greater in women.In the bile of patients with cholelithiasis the concentration of total bile acids was reduced in both sexes but there was no sex difference in the molar percent of any of the bile acids. The molar percent of CDCA (both glycine and taurine conjugates) was reduced in women, while the molar percent of CA (only the glycine conjugate) was reduced in men.     

Characterisation of the Glycine Modulatory Site of the N-Methyl-d-Aspartate Receptor-Ionophore Complex in Human Brain

[3H]Glycine binding and glycine modulation of [3H]MK-801 binding have been used to study the glycine allosteric site associated with the N-methyl-D-aspartate receptor complex in postmortem human brain. The effect of glycine on [3H]MK-801 binding appeared sensitive to duration of terminal coma, and possibly postmortem delay. Thirty percent of the binding occurred in a subfraction of brain tissue and did not show enhancement by glycine and glutamic acid. [3H]Glycine binding to a subfraction free from this component was studied and showed high specific binding. KD and Bmax values showed considerable intersubject variability which did not appear to be due to demographic features or to tissue content of amino acids with an affinity for this site. The pharmacological characteristics of binding in this subfraction and a correlation between Bmax values and the maximal enhancement of [3H]MK-801 binding by glycine are consistent with [3H]glycine binding occurring to an N-methyl-D-aspartate receptor complex associated site. Further support for this is provided by a significantly lower Bmax value for [3H]glycine binding in subjects with Alzheimer's disease and reduced glycine enhancement of [3H]MK-801 binding. However, the effect of perimortem factors makes it difficult to confidently attribute this solely to a disease-related change in the receptor. The possible role of the glycine allosteric site in the treatment of neuropsychiatric disorders is discussed.     

Exogenous glycine partially attenuates homocysteine-induced apoptosis and membrane peroxidation in chick embryos

The effects of exogenous glycine on homocysteine (HoCys)-induced reductions in chick (Gallus gallus) embryo viability, HoCys-induced increases in brain and hepatic membrane lipid peroxidation, HoCys-induced apoptosis (caspase-3 activities) in brain and hepatic tissues, and HoCys-induced reductions in brain and hepatic S-adenosylemethionine (SAM)/S-adenosylhomocysteine (SAH) levels were studied. Exogenous HoCys caused reductions in percent living embryos and reductions in embryo masses. Exogenous glycine attenuated these HoCys-induced reductions in embryo viability. Brain and liver tissues of HoCys-treated embryos exhibited increased caspase-3 activities, increased lipid hydroperoxide (LPO) levels, and reduced levels of long-chain polyunsaturated membrane fatty acids. While exogenous glycine attenuated HoCys-induced changes in brain caspase-3 activities, brain LPO levels, and brain membrane PUFA levels, exogenous glycine was less effective in attenuating HoCys-induced changes in hepatic caspase-3 activities and hepatic membrane PUFA levels. HoCys-induced reductions in SAM/SAH ratios were observed in brains and livers. Exogenous glycine attenuated HoCys-induced reductions in brain SAM/SAH. However, glycine was unable to attenuate HoCys-induced reductions in hepatic SAM/SAH levels.     

Effect of sludge type on poliovirus association with and recovery from sludge solids

Sludge type was found to affect the degree of association between seeded poliovirus type 1 (LSc) and sludge solids. The mean percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested sludges was 57.2, 70.4, and 94.7, respectively. The degree of association between poliovirus and sludge solids was significantly greater for aerobically digested sludges than for the other two sludge types. Sludge solids associated viruses were eluted using 0.05 M glycine buffer, pH 10.5-11.0, and subsequently concentrated by organic flocculation. The effectiveness of the glycine method in the recovery of solids-associated viruses was also found to be affected by sludge type. Significantly lower mean poliovirus recovery was found for aerobically digested sludges (14.5%) than for mixed liquors or anaerobically digested sludges (72.3 and 60.2%, respectively). The eluent used in the method was not as effective in dissociating the virus from aerobic sludge solids as it was for the other two sludge types. All other virus adsorption-elution steps of the method (i.e., virus concentration steps) were equally effective in poliovirus recovery for all three sludge types. It is suggested that future methods developed for the recovery of viruses from sludges be evaluated for the various sludge types likely to be tested.     

Molecular vibrations and normal modes in L-prolyl-glycyl-glycine using Wilson GF matrix method

Collagen is one of the most important proteins containing mostly proline hydroxyproline and glycine. In collagen, approximately 33 percent of the amino acid residues are glycine and they occur at every third position, whereas remaining percentage is constituted by mainly proline or hydroxyproline and some part by alanine etc. having no definite positional placement in the chain. Thus, a study of conformation of proline and glycine containing dipeptides and tripeptides is important for understanding the conformation of collagen as a sequence of its constituent amino acids. In the present communication, we have studied spectral features of L-proline, L-prolyl-glycine (PG), L-prolyl-alanine (PA), L-glycylglycine (GG), Collagen and L-prolyl-glycyl-glycine (PGG). We have carried out detailed normal mode analysis of only PGG, because interpretation of spectra of other proline and glycine containing peptides can be treated as derivatives of this molecule. Urey-Bradley force field, which involves non-bonded interactions in the gem and cis configurations is used for calculation of normal modes. The "best-fit" set of constants are generated for PGG.     

Human luteinizing hormone. Isolation and characterization of the native hormone and its alpha and beta subunits.

A new procedure is described for the isolation of the alpha and beta chains of the hormone. In this method, thenative hormone is incubated in acidic urea and the chains are then separated by ion-exchange chromatography. The amino-terminal residue of the alpha subunit is valine. The carboxy-terminal end of the alpha subunit is of variable length. No amino-terminal residue was detected for the beta chain; glycine was found at its carboxy-terminal end by the selective titration method. The amino acid and carbohydrate compositions of the hormone and both subunits are presented. The beta chain contains sialic acid and is devoid of galactosamine in contrast to the beta subunits of other species. Contamination of our human lutenizing hormone preparation by other pituitary glycoprotein hormones such as thyroid-stimulating hormone and follicle-stimulating hormone amounted to 0.5 and 0.25 percent by weight respectively. Cross-contamination of the initial alpha and beta subunit preparations was measured by specific radioimmunoassays and amounted to 4.1 and 2 percent by weight respecitively. Further extensive purification of these subunit preparations was then performed by means of affinity chromatography using immunosorbants. The final preparations exhibited a residual cross-contamination amounting to 0.2 and 0.02 percent by weight for the alpha and beta subunits respectively.     

Benzoic acid conjugation in tuatara

The excretion and metabolism of orally administered [¹⁴C]-labelled benzoic acid (100 mg/kg) was examined in the reptile Sphenedon punctatus (tuatara). The major excreted metabolite was chromatographically and electrophoretically identical with ornithuric acid. Conjugation with glycine or glucuronic acid was not detected. 7–21 percent of the dose was recovered from the urine and faeces, the bulk of the excreted radioactivity being eliminated in the first seven days. Free benzoic acid and conjugates were excreted in the first week but only conjugates could be detected in fauces collected at later intervals. These results are discussed in relation to the taxonomic position of tuatara.     

Pattern of repeating aromatic residues in synexin. Similarity to the cytoplasmic domain of synaptophysin

Synexin was isolated from bovine liver by high resolution cation exchange chromatography and fragmented with cyanogen bromide or trypsin. Peptides were isolated and their amino acid sequences partially determined. Twenty percent of the synexin sequence was determined in one contiguous sequence of 61 residues and a nonoverlapping sequence of 20 residues. The sequence is characterized by a hexapeptide repeat of the form YPXXXX occurring eight times in series, with phenylalanine substituting for tyrosine in two positions. The intervening amino acids (X) are predominantly proline, glycine and alanine. This pattern of periodic aromatic residues suggests the presence of a novel secondary structure and is similar to repeats present in synaptophysin, gliadin and type II keratin.     

Amino Acid Changes in a Genetic Strain of Epileptic Beagle Dogs

A neurochemical evaluation of beagle dogs with naturally occurring spontaneous generalized convulsive seizures was performed. Amino acid profiles of serum, cerebrospinal fluid (CSF), and biopsied cerebral cortex from epileptic dogs were compared with those from seizure-free siblings. No differences in absolute levels were noted. However, when levels were normalized as a percent of total free amino acids, seizures was performed. Amino acid profiles of serum, cerebrospinal fluid (CEF), and biopsied cerebral cortex from epileptic dogs were compared with those seizure-free siblings. No differences also the two groups differed in certain respects. Ten significant correlations between amino acid pairs appeared in epileptic dogs, but only one was seen in seizure-free animals. Seven of these ten correlations involved glutamate or taurine. It was noted that the highly correlated amino acids (taurine, glutamate, glycine, glutamine, alanine) all utilize sodium-dependent membrane transport processes. The sum of glutamate, aspartate, and glycine levels (competing sodium-dependent high-affinity systems) was significantly lower in epileptic beagles. Since this difference was noted in serum but not CSF or brain, it may indicate a diminished capacity of sodium-dependent high-affinity renal transport for acidic and certain small neutral amino acids.     

Determination of lipoprotein-lipase activity in human skeletal muscle tissue.

An in vitro assay system was developed for the determination of lipoprotein-lipase activity in 10--30-mg specimens of human skeletal muscle tissue. The reaction medium of the assay was based on a glycine buffer of pH 8.3 (at 37 degrees C) with a heparin concentration of 1.5 g/l (about 180 IU/ml). The enzyme activity was measured as the release of [3H]oleic acid from a serum-activated, triglyceride emulsion, in which [3H]trioleate was used as trace substance. The enzyme activity studied had the characteristic properties of lipoprotein-lipase activity, i.e. it was activated by the addition of serum or apolipoprotein C-II and inhibited in the presence of high ionic strength, protamine sulphate or apolipoprotein C-III. A mean Km of 0.40 +/- 0.13 (S.D.) mmol/l for triglyceride substrate was found in tissue samples that had very different concentrations of lipoprotein-lipase activity. This Km was similar to the low fasting concentrations of very low density lipoprotein triglycerides often found in healthy individuals. The lipoprotein-lipase activity was not decreased freezing and storing the tissue specimens in liquid nitrogen. The within-day variation of the method was 16 percent and the between-day variation 8 percent. Muscle tissue from the vastus lateralis muscle had, on the average, a 60 percent higher concentration of lipoprotein-lipase activity than the rectus abdominis muscle in the same subject.