Purification of stearidonic acid (18:4(n-3)) and hexadecatetraenoic acid (16:4(n-3)) from algal fatty acid with lipase and medium pressure liquid chromatography

Stearidonic acid (18:4(n-3)) and hexadecatetraenoic acid (16:4(n-3)) are included in some edible marine algae such as Undaria pinnatifida and Ulva pertusa with relatively high compositions (up to 40%) of total fatty acids. In order to prepare 16:4(n-3) and 18:4(n-3) enriched fatty acid concentrates, we screened for a suitable lipase which concentrates these acids by the removal of other fatty acids in the selective esterification reaction reported by Shimada et al. (Shimada et al. (1997), J. Am. Oil Chem. Soc., 74, 1465-1470). In combination with the lipase reaction and reversed-phase medium pressure liquid chromatography, we purified 18:4(n-3) and 16:4(n-3) to more than 95% purity.     

Stearidonic acid (18:4ω3) in Primula florindae

Stearidonic acid (18:4ω3), which is reported to be of rare occurrence in the plant kingdom and which is of considerable dietary and pharmaceutical interest has been found in three closely related Primula species. It occurs, together with γ-linolenic acid (3–4% of the seed oil total fatty acids), in significant percentages in Primula florindae (11%), P. sikkimensis (14%) and P. alpicola (14%). 18:4(ω3 may also be of chemotaxonomic interest in the genus Primula, as high levels may be typical for section Sikkimensis. The only commercial plant source of stearidonic acid known so far is the seed oil of Ribes nigrum.     

Determination of Essential Fatty Acid Composition among Mutant Lines of Canola （Brassica napus）, through High Pressure Liquid Chromatography

The present study aimed to quantify the methyl esters of lenoleic acid (LA), γ-lenolenic acid (LNA) and oleic acid (OL) in the oil of Brassica napus mutants. Five stable mutants (ROO-75/1, ROO-100/6, ROO-125/12, ROO-125/14, and ROO-125/17)of B. napus cv. 'Rainbow' (P) and three mutants (W97-95116, W97-0.75/11 and W97-.075/13) of B. napus cv. 'Westar' (P) at M6 stage, exhibiting better yield and yield components, were analyzed for essential fatty acids. The highest seed yield was observed in the mutant (ROO-100/6) followed by ROO-125/14 of Rainbow, that is, 34% and 32% higher than their parent plants, respectively. Westar mutant W97-75/11 also showed 30% higher seed yield than its parent plant. High performance liquid chromatography analysis of the composition of fatty acids indicated that OL was the most dominant fatty acid, ranging from 39.1 to 66.3%; LA was second (15.3-41.6%) and LNA was third (18.1-28.9%). Mutant ROO-125/14 showed higher OL contents than parent (Rainbow). These results are expected to support the approval of ROO-125/14 in the National Uniform Varietal Yield Trials (NUVYT) as a new variety based on high oil quality.     

Downstream processing and purification of eicosapentaenoic (20:5n-3) and arachidonic acids (20:4n-6) from the microalga Porphyridium cruentum

Eicosapentaenoic acid (FPA, 20:5n-3) and arachidonic acid (AA, 20:4n-3)were obtained from the microalga Porphyridium cruentum by a three-stepprocess: fatty acid extraction by direct saponification of biomass,polyunsaturated fatty acid (PUFA) concentration by urea inclusion complexingand EPA isolation by high-performance liquid chromatography (HPLC). Twosolvents were tested for direct saponification of lipids in biomass. Themost efficient solvent, ethanol (96% v/v), extracted 75% ofthe fatty acids. PUFAs concentration by urea inclusion employed a urea/fattyacid ratio of 4:1 wt/wt at the crystallization temperatures of 4°C and28°C. Concentration factors were similar at both temperatures, but theEPA and AA recoveries were higher at 28°C (67.7% and 61.8%for the two acids, respectively). EPA and AA were purified from this PUFAconcentrate using analytical scale HPLC and the best results of thisseparation were scaled up to preparative level (4.7 i. d. × 30 cmcompression radial cartridge). A 94.3% pure EPA fraction and a81.4% pure AA fraction were obtained. Suitability of severalmicroalgae (Porphyridium cruentum, Phaeodactylum tricornutum and Isochrysisgalbana) and cod liver oil as sources of highly pure PUFAs, mainly EPA, wascompared.     

Rates and products of long-chain Fatty Acid synthesis from [1-C]acetate in chloroplasts isolated from leaves of 16:3 and 18:3 plants

Chloroplasts highly active in the synthesis of long-chain fatty acids from [1-¹⁴C]acetate were prepared from leaves of Solanum nodiflorum, Chenopodium quinoa, Carthamus tinctorius, and Pisum sativum. These preparations were used to test whether the various additions to incubation media found to stimulate the synthesis of particular lipid classes in vitro by Spinacia oleracea chloroplasts were applicable generally. Chloroplasts from 18:3 plants incorporated a greater proportion of radioactivity into unesterified fatty acids under control conditions than did those from 16:3 plants. Supplying exogenous sn-glycerol 3-phosphate or Triton X-100 to chloroplasts increased the synthesis of glycerolipids in all cases and accentuated the capacity of chloroplasts from 18:3 plants to accumulate phosphatidic acid rather than the diacylglycerol accumulated by chloroplasts from 16:3 plants. The UDP-galactose-dependent synthesis of labeled diacylgalactosylglycerol was much less active in incubations of chloroplasts from 18:3 plants also containing sn-glycerol 3-phosphate and Triton X-100 compared with similar incubations from 16:3 plants. Exogenous CoA stimulated total fatty acid synthesis in all chloroplast preparations and the further addition of ATP diverted radioactivity from the unesterified fatty acid to acyl-CoA. The results have been discussed in terms of the two pathway hypothesis for lipid synthesis in leaves.     

高速逆流色谱结合半制备液相色谱制备甘青青兰中3种活性成分

采用高速逆流色谱（HSCCC）结合液相色谱法制备甘青青兰（Dracocephalum tanguticum Maxim.）中绿原酸、胡麻甙-6″-乙酯、迷迭香酸,建立快速分离制备甘青青兰中活性成分的方法。采用半制备型高效液相色谱（SP-HPLC）富集甘青青兰乙酸乙酯萃取物中绿原酸、胡麻甙-6″-乙酯、迷迭香酸,再用制备液相（Pre-HPLC）和HSCCC对富集物进行分离纯化,获得54 mg化合物Ⅰ、130 mg化合物Ⅱ和200 mg化合物Ⅲ,纯度分别为96.9%、97.9%和95.1%,经核磁共振碳谱（¹³CNMR）和氢谱（¹HNMR）分别鉴定为绿原酸、胡麻甙-6″-乙酯和迷迭香酸。本实验方法适用于甘青青兰乙酸乙酯萃取物中绿原酸、胡麻甙-6″-乙酯和迷迭香酸的分离制备,并避免了传统分离方法操作繁多、试剂消耗量大、不可回收等弊端,为分离甘青青兰活性成分、制备对照品等研究提供了参考依据。     

Hydrolysis and Esterification with Lipase from Candida Cylindracea. Influence of the Reaction Conditions and Acid Moiety on the Enantiomeric Excess

The enantiomeric ratio for hydrolysis and synthesis of 1-phenyl ethanol esters of straight chain aliphatic carboxylic acids catalyzed by Candida cylindracea lipase was determined. A distinct maximum in enantiomeric ratio was observed for valeric and caproic acid in the hydrolytic direction. No significant maximum could be determined in the esterification reaction. Even though the enzyme provided larger enantiomeric ratios in the synthetic direction the enantiomeric excess of the alcohol was not higher. The enantiomeric excess was depressed by racemization reactions in the esterification as the reaction approached thermodynamic equilibrium at an insufficient conversion. While choosing the optimal chain length of the acyl donor is important in hydrolytic reactions it seems to be of greater value to raise the equilibrium conversion in the esterification reactions.     

Apparent relative retention of the phosphatidylethanolamine molecular species 18:0-20:5(n-3), 16:0-22:6(n-3) and the sum 16:0-20:4(n-6) plus 16:0-20:3(n-9) in the liver microsomes of pig on an essential fatty acid deficient diet.

Attempts at a better understanding of the cell membrane organization and functioning need to assess the physical properties which partly depend (i) on the positional distribution of the fatty acids in the membrane phospholipids (PLs) and (ii) on the way by which the PL molecular species are affected by exogenous fatty acids. To do that, the effects of essential (polyunsaturated) fatty acid (EFA) deficiency and enrichment were studied in the liver microsomes of piglets feeding on either an EFA-deficient diet or an EFA-enriched diet containing hydrogenated coconut oil or a mixture of soya + corn oils, respectively. After derivatization, the diacylated forms of choline and ethanolamine PLs were analyzed using a combination of chromatographic techniques and fast-atom bombardment-mass spectrometry. The dinitrobenzoyl-diacylglycerol derivatives corresponding to the molecular species of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were identified. It appears that three factors brought about a marked apparent relative retention: the nature of (i) the base of the polar head, (ii) fatty acids at the sn-1 position and (iii) fatty acids at the sn-2 position. The highest apparent relative retentions were displayed by the 18:0-20:5(n-3)-PE and 16:0-22:6(n-3)-PE. It is noteworthy that the behavior of 20:3 n-9--which is synthesized during the EFA-deficient diet by the same bioconversion system as 20:4 n-6--was very similar to that of 20:4 n-6 during the formation of PC and PE molecular species and that the molecular species of PE containing 20:4(n-6) and 20:3(n-9), gathered together as metabolical homologues, were also apparently retained, particularly in association with 16:0. Present observations are consistent with some others showing retention or preferential distribution of EFA in PE and suggest that specific acyltransferase(s), ethanolamine phosphotransferase and methyltransferase would be mainly involved for PE and PC formation in liver endoplasmic reticulum. Fast-atom bombardment-mass spectrometry of intact phospholipids enables us to show that there is no very long chain dipolyunsaturated phospholipid in liver endoplasmic reticulum.     

高效液相色谱法同时测定银凤桃中的赤霉素和脱落酸

赤霉素(GA3)和脱落酸(ABA)是果实组织中的两个重要激素,本试验用SymmetryC18色谱柱(4.6mm×150mm),以乙腈和1.8%乙酸[V(CH3CN)∶V(1.8%CH3COOH)=1∶1]为流动相,流速为0.5mL·min-1,Wa-ters2487UV-检测器,在检测波长254nm,柱温25℃的条件下,同时分离并测定了银凤桃中的GA3和ABA。GA3和ABA的分离效果理想,回收率分别达到100.1%和99.8%,该方法测量灵敏度达10-2ng·g-1,精密度RSD%<0.1。     

Determination of Picomole Amounts of Dopamine, Noradrenaline, 3,4-Dihydroxyphenylalanine, 3,4-Dihydroxyphenylacetic Acid, Homovanillic Acid, and 5-Hydroxyindolacetic Acid in Nervous Tissue After One-Step Purification on Sephadex G-10, Using High-Performance Liquid Chromatography with a Novel Type of Electrochemical Detection

A determination of dopamine (DA), noradrenaline (NA), 3,4-dihydroxyphenylalanine (DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindolacetic acid (5-HIAA) in nervous tissue is described. The method is based on a rapidly performed isolation of DA, NA, DOPA, DOPAC, HVA, and 5-HIAA from one single nervous tissue sample on small columns of Sephadex G-10, followed by reverse-phase high-performance liquid chromatography with electrochemical detection. A new type of electrochemical detector based on a rotating disk electrode (RDE) was used. The rotating disc electrode was found to be a reliable and sensitive amperometric detector with several advantages over the currently used thin-layer cells. The detector appeared very useful for routine analysis. Practical details are given for the routine use of the RDE. Brain samples containing no more than 75-150 pg (DA, DOPA, DOPAC, HVA, and 5-HIAA) or 500 pg (NA) could be reproducibly assayed with high recovery (approx. 85%) and precision (approx. 5%), without the use of internal standards. Endogenous concentrations of DA, NA, DOPA, DOPAC, HVA, and 5-HIAA were determined in eight brain structures.     

Low C18 to C20 fatty acid elongase activity and limited conversion of stearidonic acid, 18:4(n-3), to eicosapentaenoic acid, 20:5(n-3), in a cell line from the turbot, Scophthalmus maximus

The TF cell line, derived from a top predatory, carnivorous marine teleost, the turbot (Scophthalmus maximus), is known to have a limited conversion of C18 to C20 polyunsaturated fatty acids (PUFA). To illuminate the underlying processes, we studied the conversions of stearidonic acid, 18:4(n-3), and its elongation product, 20:4(n-3), in TF cells and also in a cell line, AS, derived from Atlantic salmon (Salmo salar), by adding unlabelled (25 microM), U-14C (1 microM) or deuterated (d5; 25 microM) fatty acids. Stearidonic acid, 18:4(n-3), was metabolised to 20:5(n-3) in both cells lines, but more so in AS than in TF cells. Delta5 desaturation was more active in TF cells than in AS cells, whereas C18 to C20 elongation was much reduced in TF as compared to AS cells. Only small amounts of docosahexaenoic acid (22:6(n-3)) were produced by both cell lines, although there was significant production of 22:5(n-3) in both cultures, especially when 20:4(n-3) was supplemented. We conclude that limited elongation of C18 to C20 fatty acids rather than limited fatty acyl Delta5 desaturation accounts for the limited rate of conversion of 18:3(n-3) to 20:5(n-3) in the turbot cell line, as compared to the Atlantic salmon cell line. The results can account for the known differences in conversions of C18 to C20 PUFA by the turbot and the Atlantic salmon in vivo.     

Solubilization and Purification of an α-Amino-3-Hydroxy-5-Methylisoxazole-4-Propionic Acid Binding Protein from Bovine Brain

alpha-Amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) is a selective ligand for an excitatory amino acid receptor subtype in mammalian brain. We have solubilized an AMPA binding protein from bovine brain membranes with 1% Triton X-100 in 0.5 M phosphate buffer and 20% glycerol at 37 degrees C and purified the stable binding sites using a series of chromatographic steps. Scatchard analysis of the purified preparation showed a curvilinear plot with dissociation constants of 10.6 and 323 nM and Bmax values of 670 and 1,073 pmol/mg of protein for the high- and low-affinity sites, respectively. Inhibition constants for several excitatory amino acid analogues were similar to those obtained for other membrane and solubilized preparations. Gel filtration of the soluble AMPA binding protein showed a single peak of [3H]AMPA binding activity at Mr approximately 500,000. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified AMPA binding protein showed a single major band at Mr = 110,000. Previously, we have shown that a monoclonal antibody (KAR-B1) against a frog brain kainate binding protein selectively recognizes an unknown protein in mammalian brain migrating at Mr approximately 100,000. We now show that this antibody recognizes the major component of the purified AMPA binding protein, supporting a structural similarity between the frog brain kainate binding protein and the mammalian AMPA binding protein.     

Solubilization and Partial Purification of α-Amino-3- Hydroxy-5-Methyl-4-Isoxazolepropionic Acid Binding Sites from Rat Brain

alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) binding sites were solubilized from rat brain membranes using 1% Triton X-100 in 0.5 M potassium phosphate buffer containing 20% glycerol. The solubilized binding sites were stable, permitting biochemical and pharmacological characterization as well as partial purification. Pharmacological and binding analyses indicated that the solubilized binding sites were similar to the membrane-bound sites. Both the solubilized and the membrane-bound preparations contained high- and low-affinity AMPA binding sites in the presence of potassium thiocyanate. A similar rank order for inhibition of [3H]AMPA binding by several excitatory amino acid analogs was obtained for the soluble and membrane-bound preparations. [3H]AMPA binding to both soluble and membrane-bound preparations was increased in the presence of potassium thiocyanate. The solubilized AMPA binding sites migrated as a single peak with gel filtration chromatography, with an Mr of 425,000. Beginning with the solubilized preparation, AMPA binding sites were purified 54-fold with ion-exchange chromatography and gel filtration. The characterization and purification of these soluble binding sites is potentially useful for the molecular characterization of this putative excitatory amino acid receptor subtype.     

Alteration in Fatty Acid Composition of Adult Rat Brain Capillaries and Choroid Plexus Induced by a Diet Deficient in n-3 Fatty Acids: Slow Recovery After Substitution with a Nondeficient Diet

Wistar rats were fed for three generations with a semisynthetic diet containing either 1.5% sunflower oil (940 mg% of C18:2n-6, 6 mg% of C18:3n-3) or 1.9% soya oil (940 mg% of C18:2n-6, 130 mg% of C18:3n-3). At 60 days of age, the male offspring of the third generation were killed. The fatty acyl composition of isolated capillaries and choroid plexus was determined. The major changes noted in the fatty acid profile of isolated capillaries were a reduction (threefold) in the level of docosahexaenoic acid and, consequently, a fourfold increase in docosapentaenoic acid in sunflower oil-fed animals. The total percentage of polyunsaturated fatty acids was close to that in the soya oil-fed rats, but the ratio of n-3/n-6 fatty acids was reduced by threefold. In the choroid plexus, the C22:6n-3 content was also reduced, but by 2.6-fold, whereas the C22:5n-6 content was increased by 2.3-fold and the ratio of n-3/n-6 fatty acids was reduced by 2.4-fold. When the diet of sunflower oil-fed rats was replaced with a diet containing soya oil at 60 days of age, the recovery in content of n-6 and n-3 fatty acids started immediately after diet substitution; it progressed slowly to reach normal values after 2 months for C22:6n-5 and 2.5 months for C22:6n-3. The recovery in altered fatty acids of choroid plexus was also immediate and very fast. Recovery in content of C22:5n-6 and C22:6n-3 was complete by 46 days after diet substitution.     

Galactolipid composition of the star-shaped dinoflagellate Asterodinium gracile (Kareniaceae): presence of hexadecatetraenoic acid (16:4(n-3))-containing monogalactosyldiacylglycerol as the predominant galactolipid and chemotaxonomic closeness to Karenia mikimotoi as the only other known Kareniacean producer of this fatty acid

Asterodinium gracile is a morphologically distinct, star-shaped member of the Kareniaceae with, like canonical Kareniaceae, a tertiary plastid of haptophyte origin. However, A. gracile's complement of carotenoid photosynthetic pigments has been shown to be chemotaxonomically atypical in that it possesses much less fucoxanthin when compared to that of other, canonical Kareniaceae in the genera Karenia, Karlodinium, and Takayama, also with a tertiary plastid of haptophyte origin. To date, Karenia mikimotoi, Karenia papilionacea, and Karenia selliformis are the only canonical Kareniaceae that have been shown to have a chemotaxonomically atypical carotenoid pigment composition in that they possess a gyroxanthin diester-like carotenoid not observed in other species of Karenia, Karlodinium, or Takayama (recognizing that Karenia, in general, produces fucoxanthin derivatives not observed in Karlodinium or Takayama). As a photosynthetic organism, K. mikimotoi has been shown to resemble Karenia brevis such that both species possess the chloroplast-associated galactolipids mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively) enriched with octadecapentaenoic acid (18:5(n-3)) in the sn-1 position, and hexadecenoic acid (16:0) and tetradecanoic acid (14:0) at the sn-2 position. However, K. mikimotoi is chemotaxonomically atypical beyond its carotenoid composition in that it possesses MGDG and DGDG with hexadecatetraenoic acid (16:4(n-3)), which has not been observed in any other members of the Kareniaceae, in the sn-2 position as major galactolipids. The goal of this study was to characterize the galactolipids of A. gracile with the hypothesis that they would also be atypical when compared to other canonical Kareniaceae because of A. gracile's atypical carotenoid pigment composition. To this end, we report that like K. brevis and K. mikimotoi, A. gracile produces MGDG and DGDG enriched in 18:5(n-3) at the sn-1 position and C₁₄ fatty acids, such as 14:0, at the sn-2 position, and like K. mikimotoi, it produces 18:5(n-3)/16:4(n-3) MGDG, yet here as its most abundant galactolipid.     

Simultaneous Determination of 3,4-Dihydroxyphenylalanine, 5-Hydroxytryptophan, Dopamine, 4-Hydroxy-3-Methoxyphenylalanine, Norepinephrine, 3,4-Dihydroxyphenylacetic Acid, Homovanillic Acid, Serotonin, and 5-Hydroxyindoleacetic Acid in Rat Cerebrospinal Fluid and Brain by High-Performance Liquid Chromatography with Electrochemical Detection

A method using reversed-phase ion-pair high-performance liquid chromatography with electrochemical detection for the simultaneous determination of tryptophan (TRP), 3,4-dihydroxyphenylalanine (DOPA), and their metabolites in whole brain, small-brain parts, and cerebrospinal fluid of rats has been developed. The sample preparation requires only homogenization in perchloric acid and centrifugation before injection onto the column. With a LiChrosorb RP-18 (10 micrometer) column and a mobile phase consisting of a phosphate (NaH2PO4, 0.1 M)-methanol mixture with octylsulfonate (2.6 x 10(-3) M) at pH 3.35 and 26 degrees C, the separation of DOPA, dopamine, norepinephrine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 4-hydroxy-3-methoxyphenylalanine, TRP, 5-hydroxytryptophan (5-HTP), serotonin, and 5-hydroxyindoleacetic acid was achieved. The method has been applied to study the effect of alpha-monofluoromethyldopa alone and in combination with L-DOPA or L-5-HTP, on the catechol and 5-OH indole levels in brain and CSF of the rat.     

Phase behavior in multilamellar vesicles of DPPC containing ganglioside GM3 with a C18:1 sphingoid base and a 24:0 acyl chain (GM3(18,24)) observed by X-ray diffraction

Structures and phase behavior of multilamellar vesicles of 1,2-dipalmitoyl-L-phosphatidylcholine (DPPC) containing various amount of ganglioside GM3 with a C18:1 sphingoid base and a 24:0 acyl chain (GM3(18,24)) were investigated by small-angle X-ray diffraction. Below 3.5 mol% GM3 content, the phase behavior was similar to that of pure DPPC except for a slight increase of lamellar repeat distance in the L(beta'), the P(beta') and the L(alpha) phases and a decrease of the pretransition temperature. In the range of 4-12 mol% GM3 content, another phase which has larger repeat distances coexisted with the phase observed below 3.5 mol% GM3 content. This has been interpreted that the phase separation into GM3-poor phase (denoted as A-phase) and GM3-rich phase (denoted as B-phase) took place. Above 13 mol% GM3 content, the B-phase became dominant. This phase separation may be related to the formation of GM3-enriched microdomains that had been observed on the cell surfaces which express large amounts of GM3, such as murine B16 melanoma (J. Biol. Chem. 260 (1985) 13328).     

Determination of Normetanephrine, 3,4-Dihydroxyphenylethyleneglycol (Free and Total), and 3-Methoxy-4-Hydroxyphenylethyleneglycol (Free and Total) in Rat Brain by High-Performance Liquid Chromatography with Electrochemical Detection and Effects of Drugs on Regional Concentrations

A new, fast and sensitive assay for normetanephrine (NM), free and total 3,4-dihydroxyphenylethyleneglycol (DOPEG), and free and total 3-methoxy-4-hydroxyphenylethyleneglycol (MOPEG) in brain tissue is described. The method is based on high-performance liquid chromatography with electrochemical detection. Small Sephadex G 10 columns were used for prepurification. This permitted the additional isolation and quantification of tyrosine, 3,4-dihydroxyphenylalanine, noradrenaline, dopamine, 3-methoxytyramine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, and 5-hydroxyindoleacetic acid. The compounds were determined in six brain areas (striatum, cortex, hippocampus, hypothalamus, brainstem, and cerebellum). Most DOPEG and MOPEG in rat brain was present in the conjugated form, except for the cerebellum, where about 80% of MOPEG was nonconjugated. No postmortem effects on MOPEG levels were observed; a slight increase in DOPEG in certain brain areas was found in microwave-killed rats. The effects of clonidine, yohimbine, N,N-dipropyl-5,6-ADTN, and chlorpromazine on the concentrations of the five noradrenaline (NA) metabolites were determined. Free and total DOPEG and MOPEG provide similar information on NA metabolism, whereas NM (after monoamine oxidase inhibition) reflects a different type of interaction of drugs with NA metabolism. The similarity in the pattern of drug-induced changes in NA metabolism in the various brain areas suggests that adrenoreceptors mediating NA metabolism are homogeneously distributed throughout the brain.     

Life history of the individuals buried in the St. Benedict Cemetery (Prague, 15th–18th Centuries): Insights from 14C dating and stable isotope (δ13C, δ15N, δ18O) analysis

Funerary practices and bioarchaeological (sex and age) data suggest that a mortality crisis linked to an epidemic episode occurred during the fifth phase of the St. Benedict cemetery in Prague (Czech Republic). To identify this mass mortality episode, we reconstructed individual life histories (dietary and mobility factors), assessed the population's biological homogeneity, and proposed a new chronology through stable isotope analysis (δ¹³C, δ¹⁸O and δ¹⁵N) and direct radiocarbon dating. Stable isotope analysis was conducted on the bone and tooth enamel (collagen and carbonate) of 19 individuals from three multiple graves (MG) and 12 individuals from individual graves (IG). The δ¹⁵N values of collagen and the difference between the δ¹³C values of collagen and bone carbonate could indicate that the IG individuals had a richer protein diet than the MG individuals or different food resources. The human bone and enamel carbonate and δ¹⁸O values suggest that the majority of individuals from MG and all individuals from IG spent most of their lives outside of the Bohemian region. Variations in δ¹⁸O values also indicate that all individuals experienced residential mobility during their lives. The stable isotope results, biological (age and sex) data and eight ¹⁴C dates clearly differentiate the MG and IG groups. The present work provides evidence for the reuse of the St. Benedict cemetery to bury soldiers despite the funeral protest ban (1635 AD). The Siege of Prague (1742 AD) by French‐Bavarian‐Saxon armies is identified as the cause of the St. Benedict mass mortality event. Am J Phys Anthropol 151:202–214, 2013. © 2013 Wiley Periodicals, Inc.