Changes in free fatty acids of awamori during aging

The fatty acid components of awamori during aging were as follows. The total amount of volatile acids calculated as acetic acid ranged from 20 to 140 mg/l, the main acid was acetic acid, and the proportion of acetic acid to total acids ranged from 35 to 80 per cent. The main acids other than acetic acid were propionic acid and i-butyic acid. Differences were observed in fatty acid constituents between awamori and other alcoholic beverages.Certain components tended to increase during maturation in kame (porous earth-enware pots): acetic acid, i-butyric acid, i-valeric acid, valeric acid, capric acid, lauric acid, myristic acid and total fatty acids. Others, however, showed no distinct changes: propionic acid, butyric acid, caproic acid, caprylic acid, palmitic acid, stearic acid, oleic acid and linoleic acid.During maturation in non-porous containers (stainless-steel or glass-linked tanks), on the other hand, caprylic acid, capric acid, lauric acid and myristic acid components tended to increase, while no distinct changes however were shown by acetic acid, propionic acid, i-butyric, butyric acid, i-valeric acid, valeric acid, caproic acid, palmitic acid, stearic acid, oleic acid, linoleic acid and total fatty acids.     

Synthesis and evaluation of in vitro antiviral activity of novel phenoxy acetic acid derivatives

Several substituted phenoxy acetic acid derived pyrazolines were synthesized by the reaction between 2-{4-[3-(2,4-dihydroxyphenyl)-3-oxo-1-propenyl]-2-methoxyphenoxy} acetic acid and substituted acid hydrazides and were tested for their in vitro cytotoxicity and antiviral activity. None of the compounds showed any specific antiviral activity [50% antivirally effective concentration (EC₅₀) ≥ 5-fold lower than minimum cytotoxic concentration]. The most cytotoxic of the series was 2-{4-[3-(2,4-dihydroxyphenyl)-1-(2-hydroxybenzoyl-4,5-dihydro-1H-5-pyrazolyl]-2-methoxyphenoxy}acetic acid (3_j), with a minimum cytotoxic concentration of 0.16 μg/mL in human embryonic lung (HEL) cells.     

Enzymatic preparation of a carbohydrate ester of medium-chain fatty acid in supercritical carbon dioxide

Acylation of methyl--d-fructofuranoside and caprylic acid with an immobilized lipase, Novozym 435, was carried out in supercritical carbon dioxide (SCCO₂) with tert-butanol as a co-solvent. Initial rate of acylation was 12-fold higher in SCCO₂ than in tert-butanol. The equilibrium conversion was increased up to 70% with an increase in the molar ratio up to a maximum of 1:20 (methyl--d-fructofuranoside:caprylic acid) at 16 MPa and 70 °C.     

Enzymatic synthesis of short‐chain esters in n‐hexane and supercritical carbon dioxide: Effect of the acid chain length

Enzymatic synthesis is the preferred way to produce so‐called “natural products.” Hydrolases have been used for short‐chain ester synthesis. These esters present a pleasant flavor and they have a lot of applications in different industries. Novozym 435 from Candida antarctica (EC 3.1.1.3, triacylglycerol lipase) was used for hexyl ester synthesis in n‐hexane and supercritical carbon dioxide (SCCO₂). Direct esterification provided higher yields than transesterification for the synthesis of esters. Several carboxylic acids of different chain lengths were tested for the esterification reactions: acetic, propionic, butyric, caproic and caprylic acids. The reactions were carried out at 40°C and the amount of enzyme used was 13.8 g/mol alcohol. Substrates were added at equimolar concentrations, with sufficient stirring to avoid external diffusion control. Different substrate concentrations up to 1.5 M were used. The working pressure was 14 MPa in the case of SCCO₂ and atmospheric pressure in the case of organic solvent. The results in both solvents show that the reaction rate increases with the chain length of the acid, but the final yields were similar. However, some of the reactions prove to be faster in SCCO₂, except for hexyl acetate and propionate synthesis, in which acetic and propionic acid presented a lower solubility in SCCO₂ due to its high polarity. Moreover, an acetic acid concentration of 1.5 M brought about a strong inhibition of the enzyme activity.     

Nitric oxide is induced by wounding and influences jasmonic acid signaling in<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

Nitric oxide (NO) has been associated with plant defense responses during microbial attack, and with induction and/or regulation of programmed cell death. Here, we addressed whether NO participates in wound responses in Arabidopsis thaliana (L.) Heynh.. Real-time imaging by confocal laser-scanning microscopy in conjunction with the NO-selective fluorescence indicator 4,5-diaminofluorescein diacetate (DAF-2 DA) uncovered a strong NO burst after wounding or after treatment with JA. The NO burst was triggered within minutes, reminiscent of the oxidative burst during hypersensitive responses. Furthermore, we were able to detect NO in plants (here induced by wounding) by means of electron paramagnetic resonance measurements using diethyldithiocarbamate as a spin trap. When plants were treated with NO, Northern analyses revealed that NO strongly induces key enzymes of jasmonic acid (JA) biosynthesis such as allene oxide synthase (AOS) and lipoxygenase (LOX2). On the other hand, wound-induced AOS gene expression was independent of NO. Furthermore, JA-responsive genes such as defensin (PDF1.2) were not induced, and NO induction of JA-biosynthesis enzymes did not result in elevated levels of JA. However, treatment with NO resulted in accumulation of salicylic acid (SA). In transgenic NahG plants (impaired in SA accumulation and/or signaling), NO did induce JA production and expression of JA-responsive genes. Altogether, the presented data demonstrate that wounding in Arabidopsis induces a fast accumulation of NO, and that NO may be involved in JA-associated defense responses and adjustments.Abbreviations AOS Allene oxide synthase - cPTIO Carboxy-2-phenyl-4,4,5,5-tetramethylimidazolinone-3-oxide-1-oxyl - DAF-2 DA 4,5-Diaminofluorescein diacetate - DETC Diethyldithiocarbamate - EPR Electron paramagnetic resonance - iNOS Inducible nitric oxide synthase - JA Jasmonic acid - JIP Jasmonic acid-induced protein - LOX2 Lipoxygenase 2 - NO Nitric oxide - OPR3 12-Oxophytodienoate reductase - PDF1.2 Plant defensin - ROS Reactive oxygen species - SA Salicylic acid - SNP Sodium nitroprusside     

In vitro propagation of Lilium testaceum and structural investigation of the storage β-1,4-glucomannan

Bulblet and callus cultures of Lilium testaceum were initiated in vitro from bulbscales. Continous propagation of the bulblet cultures was achieved on a modified Murashige and Skoog agar medium containing 1-naphthalene acetic acid (0.1 mg/l) and kinetin (0.1 mg/l) as phytohormones. The in vitro grown bulbs synthesized large quantities of storage ß-1,4-glucomannans (mannose: glucose = 73; molecular weight = 200 kd) with an identical structure to the glucomannans from the in vivo grown bulbs. Higher 1-naphthalene acetic acid concentrations (1 mg/l) resulted in increased callus formation. Liquid suspension cultures derived from callus exhibited only small amounts of reserve glucomannans.Abbreviations DEAE 2-(diethylamino)ethyl - GF growth factor - GM glucomannan - GPC gel permeation chromatography - IAA indole-3-acetic acid - IEC ion exchange chromatography - MS Murashige and Skoog - MW molecular weight - MWCO molecular weight cut off - NAA 1-naphthalene acetic acid - NMR nuclear magnetic resonance - PVP polyvinylpyrrolidone     

Cryopreservation of embryonic axes of trifoliate orange (Poncirus trifoliata [L.] RAF.)

Halved shoot bases of Allium tuberosum Rottl. ex Spreng. proliferated both axillary and adventitious shoots on B5 medium (1968) supplemented with either 6-benzylaminopurine (0.5 mg/l) or 1-naphthalene acetic acid (0.1 mg/l) and 2-isopentenyladenine (0.5 mg/l). In vitro shoots proliferated further numerous shoots upon subculture to fresh medium, and these shoots rooted spontaneously. Plantlets were transplanted successfully to soil and retained the diploid condition of the parents.Abbreviations B5 Gamborg et al. (1968) medium - BAP 6-benzylaminopurine - 2ip 2isopentenyladenine - NAA 1-naphthalene acetic acid     

电子穿梭体对菌株Clostridium butyricum LQ25异化铁还原性质影响

【目的】在异化铁还原细菌培养体系中,通过外加电子穿梭体,分析电子穿梭体种类与浓度对细菌异化铁还原性质的影响。【方法】以一株发酵型异化铁还原细菌Clostridium butyricum LQ25为研究对象,设置水溶性介体蒽醌-2-磺酸钠和核黄素作为外加电子穿梭体。【结果】在氢氧化铁为电子受体、葡萄糖为电子供体培养条件下,不同浓度蒽醌-2-磺酸钠和核黄素对菌株LQ25异化铁还原效率影响具有显著性差异。外加蒽醌-2-磺酸钠浓度为0.5 mmol/L时,菌株累积产生Fe(Ⅱ)浓度最高,为12.95±0.08 mg/L,相比对照组提高88%。核黄素浓度为100mg/L时,菌株累积产生Fe(Ⅱ)浓度是11.06±0.04mg/L,相比对照组提高61%。外加电子穿梭体能够改变菌株LQ25发酵产物中丁酸和乙酸浓度,提高乙酸相对含量。【结论】蒽醌-2-磺酸钠和核黄素作为外加电子穿梭体能显著促进细菌异化铁还原效率,为揭示发酵型异化铁还原细菌胞外电子传递机制提供实验支持。     

Caprylic acid ameliorates rotenone induced inflammation and oxidative stress in the gut-brain axis in Zebrafish

Background

Dysfunction of the gastrointestinal tract (GIT) is one of the most common non-motor symptom of Parkinson’s Disease (PD). Pathological processes causing PD were suggested to initiate in the enteric nervous system (ENS) and proceed to the central nervous system (CNS). There are studies showing that low-carbohydrate ketogenic diets can improve motor symptoms of PD. Caprylic acid (C8) is the principal fatty acid component of the medium-chain triglycerides in the ketogenic diets. In this study, we aimed to evaluate the effects of caprylic acid, in neurotoxin exposed zebrafish focusing on the relationship between intestinal and brain oxidative stress and inflammation.

Methods

Adult zebrafish were exposed to rotenone (5 μg/L) (R group) and caprylic acid (20 and 60 mg/mL) (L?+?HDCA and R?+?HDCA groups) for 30 days. At the end of 30 days locomotor activities were determined. Levels of lipid peroxidation (LPO), nitric oxide, glutathione and superoxide dismutase and glutathione S-transferase activities were determined by spectrophotometric methods and gene expressions of tnf?, il1, il6, il21, ifn? and bdnf were evaluated by RT-PCR in the brain and intestinal tissues of zebrafish.

Results

Caprylic acid ameliorated LPO, NO, SOD and the expressions of tnf?, il1, il6, il21, ifn? and bdnf in brain and intestines. Locomotor activities were only ameliorated in high dose R?+?HDCA group.

Conclusions

Caprylic acid ameliorated the neurotoxin-induced oxidative stress and inflammation both in the brain and intestines and enhanced locomotor activity in zebrafish.

Graphical abstract

     

Effects of acetic acid and lactic acid on the growth of Saccharomyces cerevisiae in a minimal medium

Specific growth rates (μ) of two strains of Saccharomyces cerevisiae decreased exponentially (R ²>0.9) as the concentrations of acetic acid or lactic acid were increased in minimal media at 30°C. Moreover, the length of the lag phase of each growth curve (h) increased exponentially as increasing concentrations of acetic or lactic acid were added to the media. The minimum inhibitory concentration (MIC) of acetic acid for yeast growth was 0.6% w/v (100 mM) and that of lactic acid was 2.5% w/v (278 mM) for both strains of yeast. However, acetic acid at concentrations as low as 0.05–0.1% w/v and lactic acid at concentrations of 0.2–0.8% w/v begin to stress the yeasts as seen by reduced growth rates and decreased rates of glucose consumption and ethanol production as the concentration of acetic or lactic acid in the media was raised. In the presence of increasing acetic acid, all the glucose in the medium was eventually consumed even though the rates of consumption differed. However, this was not observed in the presence of increasing lactic acid where glucose consumption was extremely protracted even at a concentration of 0.6% w/v (66 mM). A response surface central composite design was used to evaluate the interaction between acetic and lactic acids on the specific growth rate of both yeast strains at 30C. The data were analysed using the General Linear Models (GLM) procedure. From the analysis, the interaction between acetic acid and lactic acid was statistically significant (P≤0.001), i.e., the inhibitory effect of the two acids present together in a medium is highly synergistic. Journal of Industrial Microbiology & Biotechnology (2001) 26, 171–177. Received 06 June 2000/ Accepted in revised form 21 September 2000     

Pollination-induced senescence of Phalaenopsis petals

The longevity of cut Phalaenopsis (Phalaenopsis hybrid, cv. Herbet Hager) flowers is normally 2 to 3 weeks. After pollination however, there was a rapid acceleration of the wilting process, beginning after only 24 h. Enhancement of senescence in several Phalaenopsis cultivars as well as in Doritaenopsis, Dendrobium and Cymbidium, was induced by successful pollination and only slightly or not at all by emasculation. Wilting of the flowers was accompanied by a loss of water from cells of the upper layer of the petals, leading to their upward folding. Following pollination there was an increase in ethylene production and sensitivity to ethylene. The increase in ethylene production began about 10 h after pollination and reached its peak after 30 h. An obvious increase in sensitivity to ethylene could already be detected 4 h after pollination and reached its peak 10 h after pollination. The increase en ethylene sensitivity following pollination was not dependent on endogenous ethylene production as it occurred also in flowers treated with (aminooxy)acetic acid, an inhibitor of ethylene biosynthesis.Abbreviations AOA = (aminooxy)acetic acid - RH = relative humidity - SEM = scanning electron microscope     

Single-cell protein of Rhodotorula sp. Y-38 from ethanol, acetic acid and acetaldehyde

Summary Continuous cultivation of Rhodotorula sp. Y-38 was carried out on ethanol, acetic acid or acetaldehyde. At a feed concentration of 1.0 % (w/v) ethanol, the cell yield of 64 g/100 g ethanol and crude protein of 52 g/100 g biomass were obtained at D=0.5 h^-1. The respective value of the content of amino acids and nucleic acids was 42.6 and 9.4 g/100 g biomass. At 2.0 % (w/v) acetic acid, cell yield was found to be 50 g/100 g acetic acid at D=0.4 h^-1. The optimum dilution rate ranged between 0.3 and 0.4 h^-1. At 0.05 % (w/v) acetaldehyde, the maximum cell yield was obtained at D=0.14 h^-1.     

The ocs element in the soybean GH2/4 promoter is activated by both active and inactive auxin and salicylic acid analogues

The octopine synthase (ocs or ocs-like) element has been previously reported to be responsive to the plant hormones, auxin, salicylic acid, and methyl jasmonate. Using transient assays with carrot protoplasts, we have demonstrated that an ocs element from the soybean auxin-inducible GH2/4 promoter is not only activated by strong auxins (i.e, 2,4-dichlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic acid, -naphthalene acetic acid) and salicylic acid, but also by weak auxin analogues (-naphthalene acetic acid), inactive auxin analogs (i.e., 2,3-dichlorophenoxyacetic acid, 2,4,6-trichlorophenoxyacetic acid), and inactive salicylic acid analogs (3-hydroxybenzoic acid and 4-hydroxybenzoic acid). Our results indicate that the ocs element in the GH2/4 promoter is not selectively induced by plant hormones and might function similarly to tandem AP-1 sites in some animal glutathione S-transferase (GST) genes. The ocs element, like the AP-1 sites in animal GST promoters, may be induced not only by certain hormones but also by some non-hormonal stress-inducing or electrophilic agents.Abbreviations GST glutathione S-transferase - MUG 4-methyl-umbelliferyl-glucuronide - GUS -glucuronidase - 2,4-D 2,4-dichlorophenoxyacetic acid - 2,3-D 2,3-dichlorophenoxyacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - 2,4,6-T 2,4,6-trichlorophenoxyacetic acid - NAA naphthalene acetic acid - SA salicylic acid - SARE putative salicylic acid-responsive element - BA benzoic acid - UTR untranslated region - nos nopaline synthase - ocs octopine synthase - mas mannopine synthase - ocs element-(–)46 CaMV 35S promoter-GUS reporter gene: the ocs element fused to a minimal –46 cauliflower mosaic virus 35S promoter fused to a GUS reporter gene with a 3 nos untranslated region     

Studies on dehydro- -ascorbic acid 2-arylhydrazone 3-oximes: Conversion into substituted triazoles and isoxazolines

-threo-2,3-Hexodiulosono-1,4-lactone 2-(arylhydrazones) (2) were prepared by condensation of dehydro- -ascorbic acid with various arylhydrazines. Reaction of 2 with hydroxylamine gave the 2-(arylhydrazone) 3-oximes (3). On boiling with acetic anhydride, 3 gave 2-aryl-4-(2,3-di-O-acetyl- -threo-glycerol-l-yl)-1,2,3-triazole-5-carboxylic acid 5,4¹-lactones (4). On treatment of 4 with liquid ammonia, 2-aryl-4-( -threo-glycerol-l-yl)-1,2,3-triazole-5-carboxamides (5) were obtained. Acetylation of 5 with acetic anhydride-pyridine gave the triacetates, and vigorous acetylation with boiling acetic anhydride gave the tetraacetyl derivatives. Periodate oxidation of 5 gave the 2-aryl-4-formyl-1,2,3-triazole-5-carboxamides (8), and, on reduction, 8 gave the 2-aryl-4-(hydroxymethyl)-1,2,3-triazole-5-carboxamides, characterized as the monoacetates and diacetates. Controlled reaction of 2 with sodium hydroxide, followed by neutralization, gave 3-( -threo-glycerol-l-yl)-4,5-isoxazolinedione 4-(arylhydrazones), characterized by their triacetates. Reaction of 2 with HBr-HOAc gave 5-O-acetyl-6-bromo-6-deoxy- -threo-2,3-hexodiulosono-1,4-lactone 2-(arylhydrazones); these were converted into 4-(2-O-acetyl-3-bromo-3-deoxy- -threo-glycerol-l-yl)-2-aryl-1,2,3-triazole-5-carboxylic acid 5,4¹-lactones on treatment with acetic anhydride-pyridine.     

Fine-tuning ethanol oxidation pathway enzymes and cofactor PQQ coordinates the conflict between fitness and acetic acid production by Acetobacter pasteurianus

The very high concentrations required for industrial production of free acetic acid create toxicity and low pH values, which usually conflict with the host cell growth, leading to a poor productivity. Achieving a balance between cell fitness and product synthesis is the key challenge to improving acetic acid production efficiency in metabolic engineering. Here, we show that the synergistic regulation of alcohol/aldehyde dehydrogenase expression and cofactor PQQ level could not only efficiently relieve conflict between increased acetic acid production and compromised cell fitness, but also greatly enhance acetic acid tolerance of Acetobacter pasteurianus to a high initial concentration (3% v/v) of acetic acid. Combinatorial expression of adhA and pqqABCDE greatly shortens the duration of starting-up process from 116 to 99 h, leading to a yield of 69 g l^-1 acetic acid in semi-continuous fermentation. As a final result, average acetic acid productivity has been raised to 0.99 g l^-1 h^-1, which was 32% higher than the parental A. pasteurianus. This study is of great significance for decreasing cost of semi-continuous fermentation for producing high-strength acetic acid industrially. We envisioned that this strategy will be useful for production of many other desired organic acids, especially those involving cofactor reactions.     

In vitro somatic embryogenesis ofDalbergia sissoo Roxb. —a multipurpose timber-yielding tree

Somatic embryogenesis was achieved in callus cultures dervied from 40-day-old semimature zygotic embryos ofDalbergia sissoo on semi-solid Murashige and Skoog (MS) salts and vitamins supplemented with 0.46–1.16 M kinetin, 6.78–9.04 M 2,4-dichlorophenoxy acetic acid (2,4-D) and 30 g/1 sucrose. Somatic embryos proliferated rapidly by secondary somatic embryogenesis after transfer to half-strength basal MS medium supplemented with 0.46-1.16 M kinetin and 6.78–9.04 M 2,4-D with 2% (w/v) sucrose. The light-green somatic embryos germinated on half-strength MS salts and vitamins supplemented with 0.5 mg/1 abscisic acid and 2% (w/v) sucrose. The developmental stages of somatic embryogenesis were studied by light and scanning electron microscopy.Abbreviations ABA Abscisic acid - BA 6-benzyladenine - Kn kinetin - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - MS Murashige and Skoog (1962) basal medium     

Marked Synergistic Bactericidal Effects and Mode of Action of Medium-Chain Fatty Acids in Combination with Organic Acids against Escherichia coli O157:H7

The aim of this study was to examine the synergistic bactericidal effects of medium-chain fatty acids (MCFAs; caprylic, capric, and lauric acid) and organic acids (OAs; acetic, lactic, malic, and citric acid) against Escherichia coli O157:H7 and to identify their underlying mechanism(s) of action. E. coli O157:H7 was treated with MCFAs, OAs, or different combinations of MCFAs and OAs. Membrane damage and cell morphology were examined by flow cytometry and transmission electron microscopy, respectively. Combined treatment resulted in an additional log-unit reduction compared with the sum of the reductions obtained after individual treatment. For example, caprylic acid (1.0 mM, or 0.016%) and citric acid (1.0 mM, or 0.012%) alone showed negligible bactericidal effects (0.30- and 0.06-log-unit reductions, respectively); however, a marked synergistic effect (>7.15-log-unit reduction) was observed when the two were combined. Although flow cytometry and microscopic analyses of bacteria treated with individual MCFAs and OAs showed evidence of membrane disruption, the bacteria were still able to form colonies; thus, the cell damage was recoverable. In contrast, cells exposed to combined treatments showed clear membrane disintegration and/or cell death (irreversible damage). The mechanism underlying the antimicrobial effects of combined treatment with MCFAs or OAs may involve disruption of the bacterial membrane, which then facilitates the entry of other antimicrobial compounds into the cytoplasm. The main advantage of combined treatment with very low concentrations of natural antimicrobial compounds is that it is very cost-effective. Thus, this approach may be an alternative to more conventional antimicrobial treatments, such as those currently used in public health, medical centers, and the food industry.     

Immunocytochemical localization of phenolic compounds in pollen walls using antibodies againstp-coumaric acid coupled to bovine serum albumin

Summary Two different antibodies against bovine serum albumin (BSA)-p-coumaric acid-conjugates were produced and used to localize phenolic compounds in exines of pollen from different species,p-Coumaric acid (pC) was coupled to BSA either via the carboxy group (BSA-pC) or directly to the aromatic ring system (BSA-azopC). The polyclonal antibodies raised in rabbits were characterized by ELISA with homologous and heterologous antigens using turkey ovalbumin as carrier protein. The results showed that the two immune sera directed against BSA-pC and BSA-azo-pC, respectively, were specific forp-coumaric acid and structurally similar compounds. Only a very poor binding by acetic acid-ovalbumin-conjugates and no binding by turkey ovalbumin was detectable. The antibodies reacted with partially purified pollen walls and with highly purified exines. The intensity of the immune reaction was proved to be dependent upon the pollen source and the preparation of the pollen walls. Using light and electron microscopy, it was shown for the first time that, in the exines ofCucurbita maxima, antibody binding was predominantly observed in the region of the germ pore apertures, the outer foot layers, and in the micro- and macrospines. We conclude from this and other earlier published data that phenols are important structural compounds of sporopollenin.Abbrevations AA acetic acid - BA benzoic acid - BSA bovine serum albumin - BSA-azo-pC p-coumaric acid coupled in meta position to BSA by a diazo reaction - BSA-azo-pC I immune serum against BSA-azo-pC - BSA-pC p-coumaric acid coupled to BSA via the COOH-group - BSA-pC I immune serum against BSA-pC - FA ferulic acid - OVA ovalbunin from turkey - pC p-coumaric acid - pHY p-hydroxybenzoic acid - SA sinapic acid - SYA syringic acid - TAT TBS-azide-Tween-buffer - TFA trifluoroacetic acid - VA vanillic acid     

Metabolism of ferulic acid by a Penicillium sp.

Ferulic acid was metabolised by a Penicillium sp. (probably P. rubrum) by a novel route involving a preliminary demethylation to caffeic acid, followed by side-chain shortening to yield protocatechuic acid which was subsequently broken down via the ortho-fission pathway.Abbreviations used BAW = n-butanol, acetic acid, water [BAW]-(454:10:22) - BzAc = benzene, acetic acid, water [BzAc](125:72:3) - BzDA = benzene, dioxane, acetic acid [BzDA](90:25:4)