Sensitive determination of D-lactic acid and L-lactic acid in urine by high-performance liquid chromatography-tandem mass spectrometry

d ‐Lactic acid in urine originates mainly from bacterial production in the intestinal tract. Increased d ‐lactate excretion as observed in patients affected by short bowel syndrome or necrotizing enterocolitis reflects d ‐lactic overproduction. Therefore, there is a need for a reliable and sensitive method able to detect d ‐lactic acid even at subclinical elevation levels. A new and highly sensitive method for the simultaneous determination of l ‐ and d ‐lactic acid by a two‐step procedure has been developed. This method is based on the concentration of lactic acid enantiomers from urine by supported liquid extraction followed by high‐performance liquid chromatography–tandem mass spectrometry. The separation was achieved by the use of an Astec Chirobiotic? R chiral column under isocratic conditions. The calibration curves were linear over the ranges of 2–400 and 0.5–100 µmol/L respectively for l ‐ and d ‐lactic acid. The limit of detection of d ‐lactic acid was 0.125 µmol/L and its limit of quantification was 0.5 µmol/L. The overall accuracy and precision were well within 10% of the nominal values. The developed method is suitable for production of reference values in children and could be applied for accurate routine analysis. Copyright © 2011 John Wiley & Sons, Ltd.     

Poly(lactic acid)/coplasticized thermoplastic starch blend: Effect of plasticizer migration on rheological and mechanical properties

Polylactic acid (PLA) and thermoplastic starch (TPS) are known as bio‐based and biodegradable thermoplastic polymers that can be used in different applications owing to their inherent physical and mechanical properties. In order to reduce the higher costs of PLA and tuning its physical and mechanical properties suitable for short life packaging applications, blending of PLA with the TPS, more economical biodegradable polymer, has been considered in academic and industrial researches. However, melt blending of PLA with TPS without compatibilization process caused some drawbacks such as coarsening morphology and declining mechanical properties and ductility because of thermodynamic immiscibility, which may restrict its usage in packaging applications. Subsequently, our approach in this research is compatibilization of PLA/TPS blends by utilization of primary well tuning of TPS formulation with a combination of sorbitol and glycerol plasticizers. In this work, the wide composition range of melt mixed PLA/TPS blends was prepared using a laboratory twin screw extruder. The effects of microstructure on the rheological and mechanical properties of PLA/TPS blends were studied using different methods such as scanning electron microscopy (SEM) images, contact angle, oscillatory shear rheological measurements, and tensile and impact strength mechanical tests. The rheological and mechanical properties were interpreted according to the morphological features and considering the possibility of plasticizer migration from TPS to PLA phase during melt blending. Reduction in complex viscosity and storage modulus of PLA matrix samples indicates the improved melt processability of blends. Finally, in comparison with mechanical results reported in literature, our simple approach yielded the blends with elastic modulus and ductility comparable with those of chemically compatibilized PLA/TPS blends.     

Lactic acid production from cellulosic material by synergetic hydrolysis and fermentation

The hydrolysis process on corncob residue was catalyzed synergetically by the cellulase from Trichoderma reesei and the immobilized cellobiase. The feedback inhibition to cellulase reaction caused by the accumulation of cellobiose was eliminated efficiently. The hydrolysis yield of corncob residue was 82.5%, and the percentage of glucose in the reducing sugar reached 88.2%. The glucose in the cellulosic hydrolysate could be converted into lactic acid effectively by the immobilized cells of Lactobacillus delbrueckii. When the enzymatic hydrolysis of cellulose and the fermentation of lactic acid were coupled together, no glucose was accumulated in the reaction system, and the feedback inhibition caused by glucose was also eliminated. Under the batch process of synergetic hydrolysis and lactic acid fermentation with 100 g/L of cellulosic substrate, the conversion efficiency of lactic acid from cellulose and the productivity of lactic acid reached 92.4% and 0.938 g/(L·h), respectively. By using a fed-batch technique, the total concentration of cellulosic substrate and lactic acid in the synergetic process increased to 200 and 107.5 g/L, respectively, whereas the dosage of cellulase reduced from 20 to 15 IU/g of substrate in the batch process. The results of the bioconversion of renewable cellulosic resources were significant.     

Physical characterization of coupled poly(lactic acid)/starch/maleic anhydride blends plasticized by acetyl triethyl citrate

Acetyl triethyl citrate (ATC) was used as a plasticizer for poly(lactic acid) (PLA)/starch blends coupled with maleic anhydride and an initiator of 2,5-bis(tert-butylperoxy)-2,5-dimethylhexane (L101). Elongation of the blend at break was markedly increased when the ATC content was above the 8% loading level, which is referred to as the percolation threshold. The extended elongation was achieved at the expense of tensile strength and elastic modulus. Thermal transitions of the blend, including the glass transition temperature (T(g)), cold crystallization temperature (T(c)) and melting temperature (T(m)), decreased with ATC content. Thermally induced ATC migration affected the thermal behavior of the plasticized blends and reduced elongation and tensile strength, whereas the elasticity modulus increased. ATC migration increased with ambient temperature, which was controlled by the activation energy of the blend system. Leaching of ATC was slow at room temperature in distilled water, but significant in boiling water. Additionally, the leaching rate was also directly proportional to the ATC content of the blend. Glass transition temperatures of PLA/starch/MA/L101 blends with various acetryl triethyl citrate contents.     

Effects of nutrient supplements on simultaneous fermentation of nisin and lactic acid from cull potatoes

The feasibility of using cull potatoes as substrate for the simultaneous production of nisin, a natural food preservative, and lactic acid, a raw material for biopolymer production, was studied. Cull potatoes are potato tubers unacceptable for food processing because ofsize or damage caused by bruising or disease. Although cull potatoes are enriched in various nutrients including starch, minerals, and proteins, they alone still cannot provide enough essential nutrients for the growth and metabolism of Lactococcus lactis subsp. lactis (ATCC 11454). Stimulation of bacterial growth, nisin biosynthesis, as well as lactic acid production was observed when additional nutrients such as yeast extract, peptone from meat, peptone from soy (PS), corn steep solid (CSS), and distillers’ dried grains with solubles were provided. Considering the cost and availability, PS and CSS were selected as nutrient supplements for nisin and lactic acid coproduction. The conditions for nisin biosynthesis and lactic acid coproduction by L. lactis subsp. lactis in a cull potato-based medium were subsequently optimized using a statistically based experimental design.     

Synthesis of star‐shaped poly(D,L‐lactic acid‐alt‐glycolic acid)‐b‐poly(L‐lactic acid) with the poly(D,L‐lactic acid‐alt‐glycolic acid) macroinitiator and stannous octoate catalyst

Two types of three‐arm and four‐arm, star‐shaped poly(D,L ‐lactic acid‐alt‐glycolic acid)‐b‐poly(L ‐lactic acid) (D,L ‐PLGA50‐b‐PLLA) were successfully synthesized via the sequential ring‐opening polymerization of D,L ‐3‐methylglycolide (MG) and L ‐lactide (L ‐LA) with a multifunctional initiator, such as trimethylolpropane and pentaerythritol, and stannous octoate (SnOct₂) as a catalyst. Star‐shaped, hydroxy‐terminated poly(D,L ‐lactic acid‐alt‐glycolic acid) (D,L ‐PLGA50) obtained from the polymerization of MG was used as a macroinitiator to initiate the block polymerization of L ‐LA with the SnOct₂ catalyst in bulk at 130 °C. For the polymerization of L ‐LA with the three‐arm, star‐shaped D,L ‐PLGA50 macroinitiator (number‐average molecular weight = 6800) and the SnOct₂ catalyst, the molecular weight of the resulting D,L ‐PLGA50‐b‐PLLA polymer linearly increased from 12,600 to 27,400 with the increasing molar ratio (1:1 to 3:1) of L ‐LA to MG, and the molecular weight distribution was rather narrow (weight‐average molecular weight/number‐average molecular weight = 1.09–1.15). The ¹H NMR spectrum of the D,L ‐PLGA50‐b‐PLLA block copolymer showed that the molecular weight and unit composition of the block copolymer were controlled by the molar ratio of L ‐LA to the macroinitiator. The ¹³C NMR spectrum of the block copolymer clearly showed its diblock structures, that is, D,L ‐PLGA50 as the first block and poly(L ‐lactic acid) as the second block. © 2001 John Wiley & Sons, Inc. J Polym Sci Part A: Polym Chem 40: 409–415, 2002     

Saccharification and alcohol fermentation in starch solution of steam-exploded potato

Steam explosion pretreatment of potato for the efficient production of alcohol was experimentally studied. The amount of water-soluble starch increased with the increase of steam pressure, but the amounts of methanol-soluble material and Klason lignin remained insignificant, regardless of steam pressure. The potatoes exploded at high pressure were hydrolyzed into a low molecular liquid starch, and then easily converted into ethanol by simultaneous saccharification and fermentation using mixed microorganisms: an amylolytic microorganism,Aspergillus awamori, and a fermentation microorganism,Saccharomyces cerevisiae. The maximal ethanol concentration was 4.2 g/L in a batch culture at 15 g/L starch concentration, and 3.6 g/L in a continuous culture fed the same starch concentration. In the fed-batch culture, the maximal ethanol concentration increased more than twofold, compared to the batch culture.     

Stereoblock poly(lactic acid): synthesis via solid-state polycondensation of a stereocomplexed mixture of poly(L-lactic acid) and poly(D-lactic acid)

Stereoblock poly(lactic acid) consisting of D- and L-lactate stereosequences can be successfully synthesized by solid-state polycondensation of a 1:1 mixture of poly(L-lactic acid) and poly(D-lactic acid). In the first step, melt-polycondensation of L- and D-lactic acids is conducted to synthesize poly(L-lactic acid) and poly(D-lactic acid) with a medium-molecular-weight, respectively. In the next step, these poly(L-lactic acid) and poly(D-lactic acid) are melt-blended in 1:1 ratio to allow formation of their stereocomplex. In the last step, this melt-blend is subjected to solid-state polycondensation at temperature where the dehydrative condensation is allowed to promote chain extension in the amorphous phase with the stereocomplex crystals preserved. Finally, stereoblock poly(lactic acid) having high-molecular-weight is obtained. The stereoblock poly(lactic acid) synthesized by this way shows a higher melting temperature in consequence of the controlled block lengths and the resulting higher-molecular-weight. The product characterization as well as the optimization of the polymerization conditions is described. Changes in M(w) of stereoblock poly(lactic acid) (sb-PLA) as a function of the reaction time.     

Lactic acid recovery from cheese whey fermentation broth using combined ultrafiltration and nanofiltration membranes

The separation of lactic acid from lactose in the ultrafiltration permeate of cheese whey broth was studied using a cross-flow nanofiltration membrane unit. Experiments to test lactic acid recovery were conducted at three levels of pressure (1.4, 2.1, and 2.8 MPa), two levels of initial lactic acid concentration (18.6 and 27 g/L), and two types of nanofiltration membranes (DS-5DK and DS-5HL). Higher pressure caused significantly higher permeate flux and higher lactose and lactic acid retention (p<0.0001). Higher initial lactic acid concentrations also caused significantly higher permeate flux, but significantly lower lactose and lactic acid retention (p<0.0001). The two tested membranes demonstrated significant differences on the permeate flux and lactose and lactic acid retention. Membrane DS-5DK was found to retain 100% of lactose at an initial lactic acid concentration of 18.6 g/L for all the tested pressures, and had a retention level of 99.5% of lactose at initial lactic acid concentration of 27 g/L when the pressure reached 2.8 MPa. For all the test when lactose retention reached 99–100%, as much as 64% of the lactic acid could be recovered in the permeate.     

Improving polylactide/starch biocomposites by grafting polylactide with acrylic acid--characterization and biodegradability assessment

In this article, the properties of a polylactide and starch composite (PLA/starch) and an acrylic acid grafted polylactide and starch composite (PLA-g-AA/starch) were compared. The composite containing PLA-g-AA was found to have much better dispersion and homogeneity of starch in the polymer matrix than the composite containing PLA, indicating better compatibility between the two phases. Better mechanical and thermal properties of the PLA-g-AA/starch composite, notably an increase in tensile strength and elongation at breakpoint, evidenced its superiority to the PLA/starch composite. Furthermore, the lower viscosity of PLA-g-AA/starch makes it easier to process than PLA/starch. Weight loss on exposure to a soil environment over a period of three months showed that the starch in the composites was almost completely biodegradable, even at a high degree of substitution (60 wt.-% starch). After three months in soil, a reduction in the mechanical properties of the blends was observed, especially in those with higher starch contents.     

Copolyesters of isosorbide,succinic acid,and isophthalic acid: Biodegradable,high Tg engineering plastics

Isosorbide, succinyl chloride and isophthaloyl chloride are polycondensed under various reaction conditions. The heating in bulk with or without catalysts as well in an aromatic solvent without catalyst, and polycondensation with the addition of pyridine only yield low molar mass copolyesters. However, heating in chlorobenzene with addition of SnCl₂ or ZnCl₂ produces satisfactory molar masses. The number average molecular weights (M_n) of most copolyesters fall into the range of 7000–15,000 Da with polydispersities (PD) in the range of 3–9. The MALDI‐TOF mass spectra almost exclusively displayed peaks of cyclics indicating that the chain growth was mainly limited by cyclization and not by side reactions, stoichiometric imbalance or incomplete conversion. The glass‐transition temperatures increased with the content of isophthalic acid from 75 to 180 °C and the thermo‐stabilities also followed this trend. © 2013 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2013, 51, 2464–2471     

Synthesis and characterization of multiblock copolymers based on L‐lactic acid,citric acid,and poly(ethylene glycol)

Because poly(L ‐lactic acid) (PLLA) is a biodegradable polyester with low immunogenicity and good biocompatibility, it is used as a biomaterial. However, hydrophobic PLLA does not have any reactive groups. Thus, its application is limited. To increase the hydrophilicity of PLLA and accelerate its degradation rate, functionalized pendant groups and blocks were introduced through copolymerization with citric acid and poly(ethylene glycol) (PEG), respectively. This article describes the synthesis and characterization of poly(L ‐lactic‐co‐citric acid) (PLCA)‐PLLA and PLCA‐PEG multiblock copolymers. The results indicated that the hydrolysis rate was enhanced, and the hydrophilicity was improved because of the incorporation of carboxyl groups in PLCA‐PLLA. The joining of the PEG block led to improved hydrophilicity of PLCA, and the degradation rate of PLCA‐PEG accelerated as compared with that of PLCA‐PLLA. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 2073–2081, 2003     

Novel route to synthesis of allyl starch and biodegradable hydrogel by copolymerizing allyl‐modified starch with methacrylic acid and acrylamide

Maize starch was modified by allyl chloride adopting an interfacial reaction technique with cetyltrimethyl ammonium bromide as a phase‐transfer catalyst and pyridine as an acid acceptor. The degree of substitution was determined from an increasing carbon content of the modified starch. The percentage of carbon and hydrogen of the allyl‐modified starch was estimated by elemental analysis (C, H, and N), and the product characterization was done through ¹H NMR and ¹³C NMR analyses. The allyl‐modified starch was then copolymerized with methacrylic acid and a combination of methacrylic acid and acrylamide at 50 and 70 °C with potassium persulfate as an initiator. The copolymer thus formed swelled in distilled water after neutralization with sodium carbonate. The percentage of absorption capacity of the hydrogels was determined with distilled water and 0.9% NaCl solution. The highest percentage of absorption, 6500%, was achieved for the developed hydrogel containing allyl starch and acrylic monomer in a 1.7:1 w/w ratio and acrylic monomer, namely, methacrylic acid and acrylamide in a 3.2:1 w/w ratio. The study on biodegradability of the developed hydrogel showed that the hydrogel is degradable in the presence of diastase (amylase). © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 1650–1658, 2003     

Anaerobic biodegradation tests of poly(lactic acid) and polycaprolactone using new evaluation system for methane fermentation in anaerobic sludge

The anaerobic biodegradation tests of polycaprolactone (PCL) and poly(lactic acid) (PLA) powders were done at thermophilic temperature (55 °C) under aquatic conditions (total solid concentrations of the used sludge were 1.73% (undiluted sludge) and 0.86% (diluted sludge)) using a newly developed evaluation system. With this system, the evolved biogas is collected in a gas sampling bag at atmospheric pressure. This method is more convenient than using a pressure transducer or inverted graduated cylinder submerged in water. The biodegradation of PCL powder (10 g, 125–250 μm) in the diluted sludge stopped in about 47 days when the biodegradability reached 92%. The biodegradability of PLA powder (10 g, 125–250 μm) in undiluted sludge was 91% at about 75 days. The biodegradability of PLA powder (10 g, 125–250 μm) in diluted sludge was 79% at about 100 days. The biodegradability of PLA powder (5 g, 125–250 μm) in diluted sludge was 80% at about 85 days. It was found that the PCL and PLA powders were quite degraded using the new evaluation method. In addition, the smaller particle size PCL powder was biodegraded faster.     

Effect of biodegradation conditions on morphology of ternary compositions of low density polyethylene with poly(lactic acid) and starch

Polyethylene–poly(lactic acid)–starch ternary compositions were obtained in a rotor disperser under conditions of shear deformation. Their biodegradation under the action of mold fungus spores and soil was investigated using IR spectroscopy and SEM. The changes under the fungal action revealed that the ternary blends represented a nutrient medium, while the exposure to soil led to alteration in morphology due to the whole range of numerous environmental factors.     

Synthesis and Characterization of Amphiphilic Biodegradable Copolymer,Poly(aspartic acid‐co‐lactic acid)

An amphiphilic biodegradable polymer, poly(aspartic acid‐co‐lactic acid) (PAL), was synthesized by simply heating a mixture of aspartic acid (Asp) and L ‐lactide without additional catalysts or solvents. The unique branched architecture comprising succinimide units and lactic acid units was confirmed by IR and NMR spectroscopy. A copolymer of sodium aspartate and lactic acid (PALNa) was prepared by reacting PAL with an aqueous sodium hydroxide solution. The PAL was soluble in many organic solvents, while the PALNa was soluble in methanol and water. The hydrolytic degradation behavior of PAL varied with the copolymer composition. A higher Asp content resulted in a faster molecular weight decrease, and introducing glycolic acid units accelerated the degradation rate.

Microphotograph of microsphere of PAL‐1/5.     

Production of fumaric acid using rice bran and subsequent conversion to succinic acid through a two-step process

The fungal production of fumaric acid using rice bran and subsequent bacterial conversion of succinic acid using fungal culture broth were investigated. Since the rice bran contains abundant proteins, amino acids, vitamins, and minerals, it is suitable material that fungi use as a nitrogen source. The effective concentration of rice bran to produce fumaric acid was 5 g/L. A large amount of rice bran caused excessive fungal growth rather than enhance fumaric acid production. In addition, we could produce fumaric acid without the addition of zinc and iron. Fungal culture broth containing appro × 25 g/L of fumaric acid was directly employed for succinic acid conversion. The amount of glycerol and yeast extract required for succinic acid conversion was reduced to 70 and 30%, respectively, compared with the amounts cited in previous studies.     

Structure and properties of the graft copolymer of starch and p‐hydroxybenzoic acid using horseradish peroxidase

A modified starch tannage was synthesized by free radical graft copolymerization of degraded starch with p‐hydroxybenzoic acid (pHA) using horseradish peroxidase/H₂O₂ as the initiator. In this study, the effects of the degree of degradation of the starch, dosage of pHA, polymerization temperature, system pH, and horseradish peroxidase content on the tanning properties of the graft copolymer were investigated. The shrinkage temperature of leather tanned by the graft copolymer was 78 °C. The thickness increment ratio of the retanned leather was 21.6%, and meanwhile the retanned leather showed better softness. The results indicate that the graft copolymer has excellent tanning and retanning properties. The structure of the graft copolymer was analyzed by Fourier Transform Infrared Spectroscopy (FTIR), ¹Hydrogen‐Nuclear Magnetic Resonance (¹H‐NMR), and ¹³Carbon‐Nuclear Magnetic Resonance (¹³ C‐NMR) and Gel Permeation Chromatography (GPC). Compared with conventional methods, we show that a “green” leather tannage could be achieved using a radical graft copolymerization of starch and phenols. Copyright © 2011 John Wiley & Sons, Ltd.     

Water absorption and enzymatic degradation of poly(lactic acid)/rice starch composites

Poly(lactic acid) (PLA) composites consisting of PLA, rice starch (RS) (0–50 wt%) and epoxidised natural rubber (ENR50) were compounded by a twin-screw extruder and compression moulded into dumbbell specimens. Tensile tests were performed to characterize the mechanical properties of the PLA/RS composites. Morphological studies were done on the tensile fractured surface of the specimens by using scanning electron microscopy (SEM). Twenty weight percent of RS achieved a good balance of strength and stiffness. Beyond 20 wt% loading of RS, the tensile strength and elongation at break of PLA decreased drastically. This may be attributed to the agglomeration of RS, which could then act as stress concentrator. The incorporation of ENR50 increased the tensile strength and elongation at break of the PLA/RS composites remarkably, owing to the elastomeric behaviour and compatibilisation effects of ENR50. Interestingly, the morphology of PLA/RS composites transformed to a more ductile one with the addition of ENR. The kinetics of water absorption of the PLA/RS composites conforms to Fick's law. The M_m and D values are dependent on the RS and ENR concentrations. The tensile properties of the PLA/RS composites deteriorated after water absorption. The retention-ability and recoverability of the PLA/RS composites are relatively low, attributed to the hydrolysis of PLA, degradation of the PLA–RS interface and leaching of the RS particles. In addition, the tensile properties of PLA/RS composites decreased drastically upon exposure to enzymatic degradation. Extensive pinhole and surface erosion on the PLA/RS composites indicate high degree of hydrolysis. Whilst the addition of ENR leads to some improvements in tensile properties, nevertheless, it enhanced the biodegradability of the PLA/RS composites when exposed to water and -amylase enzymatic treatments.     

Production of lactic acid from food wastes

Conversion of food wastes into lactic acid by simultaneous saccharification and fermentation (SSF) was investigated. The process involves saccharification of the starch component in food wastes by a commercial amylolytic enzyme preparation (a mixture of amyloglucosidase, α-amylase, and protease) and fermentation by Lactobacillus delbrueckii. The highest observed overall yield of lactic acid in the SSF was 91% of theoretical. Lactic acid concentration as high as 80 g/L was attainable in 48 h of the SSF. The optimum operating conditions for the maximum productivity were found to be 42°C and pH 6.0. Without supplementation of nitrogen-containing nutrients, the lactic acid yield in the SSF decreased to 60%: 27 g/L of lactic acid from 60 g/L of food waste. The overall performance of the SSF, however, was not significantly affected by the elimination of mineral supplements.