Synthesis and Characterization of Poly(Butylene Sebacate-Co-Terephthalate) Copolyesters with Pentaerythritol as Branching Agent.

Poly(butylene sebacate-co-terephthalate) (PBSeT) copolyesters are prepared by melt polymerization via two-step transesterification and polycondensation using pentaerythritol (PE) as a branching agent. The effects of the incorporated PE on its chemical, thermal, mechanical, and degradation properties, along with the rheological properties of its melt, are investigated. The highest molecular weight and intrinsic viscosity along with the lowest melt flow index were achieved at a PE content of 0.2 mol%, with minimal reduction in the tensile strength and the highest tear strength. The addition of PE did not significantly influence the thermal behavior and stability of the PBSeT copolyesters; however, the elongation at break decreased with increasing PE content. The sample with 0.2 mol% PE exhibited a higher storage modulus and loss modulus as well as a lower loss angle tangent than the other samples, indicating improved melt elasticity. The incorporation of more than 0.2 mol% PE enhanced the enzymatic degradation of copolyesters. In summary, including within 0.2 mol%, PE effectively improved both the processability-related characteristics and degradation properties of PBSeT copolyesters, suggesting their potential suitability for use in agricultural and packaging materials.

A New Method to Determine Antioxidant Activities of Biofilms Using a pH Indicator (Resazurin) Model System.

Biopolymeric films were prepared with gelatin, plasticizer, and three different types of antioxidants (ascorbic acid, phytic acid, and BHA) corresponding to different mechanisms in activity. The antioxidant activity of films was monitored for 14 storage days upon color changes using a pH indicator (resazurin). The instant antioxidant activity of films was measured by a DPPH free radical test. The system using resazurin was composed of an agar, an emulsifier, and soybean oil to simulate a highly oxidative oil-based food system (AES-R). Gelatin-based films (GBF) containing phytic acid showed higher tensile strength and energy to break than all other samples due to the increased intermolecular interactions between phytic acid and gelatin molecules. The oxygen barrier properties of GBF films containing ascorbic acid and phytic acid increased due to the increased polarity, while GBF films containing BHA showed increased oxygen permeability compared to the control. According to "a-value" (redness) of the AES-R system tested with films, films incorporating BHA showed the most retardation of lipid oxidation in the system. This retardation corresponds to 59.8% antioxidation activity at 14 days, compared with the control. Phytic acid-based films did not show antioxidant activity, whereas ascorbic acid-based GBFs accelerated the oxidation process due to its prooxidant activity. The comparison between the DPPH free radical test and the control showed that the ascorbic acid and BHA-based GBFs showed highly effective free radical scavenging behavior (71.7% and 41.7%, respectively). This novel method using a pH indicator system can potentially determine the antioxidation activity of biopolymer films and film-based samples in a food system.

Grape pomace and its secondary waste management: Biochar production for a broad range of lead (Pb) removal from water.

Grape pomace (GP) management has been a challenge worldwide. We have previously demonstrated a biorefinery process to recover oil and polyphenols, and produce biofuels from GP sequentially, although over 50% of GP solid waste remains post-processing. To approach zero solid waste during GP processing, herein a pyrolysis process was designed for converting GP and its secondary processing wastes to biochars, which were then evaluated for lead (Pb) adsorption from water. GP lignin pyrolyzed at 700 °C (GPL2700 biochar) with specific surface area of 485 m2/g showed the highest Pb adsorption capacity, and achieved 66.5% of Pb removal from an initially high concentration of 300 mg/L within 30 min. At low initial Pb concentrations (50-3000 µg/L), GPL2700 biochar could reduce Pb concentrations to 0.208-77.2 µg/L. In addition, experimental and modeling results revealed that both physisorption and chemisorption mechanisms were involved in the adsorption process of GPL2700 biochar.

TEMPO-Oxidized Cellulose Nanofibril Films Incorporating Graphene Oxide Nanofillers.

To design a new system of novel TEMPO-oxidized cellulose nanofibrils (TOCNs)/graphene oxide (GO) composite, 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation was utilized. For the better dispersion of GO into the matrix of nanofibrillated cellulose (NFC), a unique process combining high-intensity homogenization and ultrasonication was adopted with varying degrees of oxidation and GO percent loadings (0.4 to 2.0 wt%). Despite the presence of carboxylate groups and GO, the X-ray diffraction test showed that the crystallinity of the bio-nanocomposite was not altered. In contrast, scanning electron microscopy showed a significant morphological difference in their layers. The thermal stability of the TOCN/GO composite shifted to a lower temperature upon oxidation, and dynamic mechanical analysis signified strong intermolecular interactions with the improvement in Young's storage modulus and tensile strength. Fourier transform infrared spectroscopy was employed to observe the hydrogen bonds between GO and the cellulosic polymer matrix. The oxygen permeability of the TOCN/GO composite decreased, while the water vapor permeability was not significantly affected by the reinforcement with GO. Still, oxidation enhanced the barrier properties. Ultimately, the newly fabricated TOCN/GO composite through high-intensity homogenization and ultrasonification can be utilized in a wide range of life science applications, such as the biomaterial, food, packaging, and medical industries.

Rapid quantification of melamine in milk using competitive 1,1'-oxalyldiimidazole chemiluminescent enzyme immunoassay.

A novel competitive 1,1'-oxalyldiimidazole (ODI) chemiluminescent enzyme immunoassay (CLEIA) was developed as a method for rapid and simple screening of melamine in milk. Fat existing in milk acts as an inhibitor in the competitive binding interaction of melamine and anti-melamine in the presence of melamine-conjugated horseradish peroxidase. Thus, the calibration curve and sensitivity of competitive ODI CLEIA for the quantification of melamine in fat free milk were wider and better than those in milk containing fat. However, a centrifuge is not a good method for removing the inhibitor because a portion of the melamine is also removed with the fat. The incubation time (20 min) for the competitive binding interaction of anti-melamine and melamine in 20% milk diluted with PBS buffer of pH 7.4 was longer than that (10 min) in 100% milk even though the sensitivity of the former was better than latter. The limit of detection (1.12 ppb) determined in rapid ODI CLEIA (dynamic range: 3.8-125 ppb) for the quantification of melamine in 20% milk not containing fat was lower than those (6.3 and 9.0 ppb) calculated in relatively time-consuming luminol CLEIA and enzyme-linked immunosorbent assay (ELISA). Also, we expect that ODI-CLEIA (dynamic range: 62.5-2000 ppb) capable of directly quantifying melamine in 100% milk without any pretreatment can be applied as a new and simple method for rapid screening of melamine in milk.

Exploration of hybrid nanocarbon composite with polylactic acid for packaging applications.

Polylactic acid (PLA) nanocomposite films were fabricated with graphene oxide (GO) and single-walled carbon nanotubes (CNT) as a hybrid-co-filler with GOCNT fraction varying from 0.05 to 0.4% by weight. The effect of the GOCNT on the physical, thermal, morphological, gas permeation, and optical properties was investigated. The X-ray diffraction test reveals no restacking and coagulation of GOCNT in the composite films. Differential Scanning Calorimetry analysis shows an insignificant shift of glass transition and melting temperature but enhanced crystallization resulting from the existence of GOCNT as a nucleating agent. Scanning Electron Microscope scans indicate GOCNT embedded homogeneously without considerable aggregates in the PLA. Transmission of ultraviolet-visible radiation decreases to 30% with increasing fraction of GOCNT while Oxygen Transmission Rate diminishes to 67% in the film. These are attributed to the tortuous pathways provided by the well-dispersed hybrid GOCNT in the PLA. Compared to the pristine PLA film, the composite film shows an increase of 75% and 130% in the tensile strength and Young's modulus, respectively. Taken together, all of these improvements observed in the hybrid GOCNT-PLA composites should provide useful guidelines in customizing designs for applications across a range of fields including packaging, life sciences, cosmetics, and conventional synthetic plastics.

Lignin-Based Biopolymeric Active Packaging System for Oil Products.

Antioxidant activity of enzymatically modified soybean protein film with two different forms of added lignin (alkali lignin and lignosulfonate) was investigated using two stimulated food systems involving direct and indirect contact with soybean oil and fish fatty acid ethyl ester (FAEE). For the direct system, control and lignin-doped films were added to oil vials which were stored at dark under 40 °C whereas for indirect, films were used to cover oil-containing glass vials stored under standard commercial lighting conditions. Autoxidation of oil samples in the direct contact system was determined by peroxide value (PV), color, headspace oxygen, and volatile compounds, while for the indirect contact system photoxidation was determined by using PV and color. For the direct contact system with soybean oil, the PV was significantly lower during storage for both lignins used compared to the control (packaging system without lignin film). There was not a significant effect of lignin on the color of the oils (P > 0.05). Modified films tested in this study did not have a significant effect on headspace oxygen contents of oil samples; however, it resulted in reduced volatile compounds for both soybean oil and fish oil samples. Based on our observation, soybean protein films with lignin showed a greater impact on soybean oil than fish oil, possibly because of high initial oxidation levels in the fish oil. Enzymatic modified soy protein films with lignin are alternative active packaging materials for highly sensitive to oxidation by radical and UV light. PRACTICAL APPLICATION: Plastic packaging materials require the use of petroleum oil and are not biodegradable. Packaging materials made from renewable, biodegradable biopolymers are of great interest but often suffer from performance problems, such as weak mechanical properties compared to petroleum-based plastics. Applying modified biopolymeric film with lignin in the inner layer of food packaging system improved some aspects of their performance during storage, not only by preventing the migration of chemical compounds from the package to the food but also by radical scavenging activity and UV-blocking ability of the packaging system.

Self-enzyme chemiluminescence immunoassay capable of rapidly diagnosing the infection of influenza A (H1N1) virus.

A highly sensitive self-enzyme immunoassay with 1,1'-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed for the first time to quantify influenza A (H1N1) virus. A specific capture antibody, capable of capturing hemagglutinin (HA) subtypes of H1N1, was immobilized on the surface of the magnetic Au-Fe3O4 nanocomposite. Neuraminidase (NA) subtype of H1N1 was acts as an enzyme in the self-enzyme immunoassay. A sample mixed with HA antibodies immobilized on the surface of magnetic Au-Fe3O4 nanocomposites was incubated for 1 h at 37 °C. After the incubation, magnetic Au-Fe3O4 nanocomposites separated with a magnetic bar were washed 3 times using phosphate buffered saline with Tween-20 (PBST). Then, 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA), a fluorogenic substrate of NA, was added and incubated for 10 min to produce 4-Methylumbelliferone (4-MU) from the enzyme reaction between MUNANA and NA of H1N1. After the incubation, the solution containing 4-MU emitted bright light with the addition of ODI-CL reagents (e.g., H2O2 and ODI). The relative CL intensity of 4-MU was proportionally enhanced with the increase of H1N1. Also, the brightness and color of 4-MU light emitted from the self-enzyme immunoassay was dependent on pH. The self-enzyme immunoassay was able to analyze trace levels of influenza A (H1N1) virus with good accuracy, precision, reproducibility and excellent selectivity. The limit of detection (LOD = 3σ/slope) was as low as 0.19 U/ml. We expect that the self-enzyme immunoassay can be applied as a cost-effective and rapid method capable of selectively quantifying a specific influenza A virus such as H1N1, H3N2, and H5N1.

Compositional Characterization of Different Industrial White and Red Grape Pomaces in Virginia and the Potential Valorization of the Major Components.

To better evaluate potential uses for grape pomace (GP) waste, a comprehensive chemical composition analysis of GP in Virginia was conducted. Eight commercial white and red pomace samples (cv. Viognier, Vidal Blanc, Niagara, Petit Manseng, Petit Verdot, Merlot, Cabernet Franc, and Chambourcin) obtained from different wineries in Virginia, USA were used. For extractives, GPs contained 2.89%-4.66% titratable acids, 4.32%-6.60% ash, 4.62%-12.5% lipids with linoleic acid being the predominant (59.0%-70.9%) fatty acid, 10.4-64.8 g total phenolic content (gallic acid equivalents)/kg GP, 2.09-53.3 g glucose/kg GP, 3.79-52.9 g fructose/kg GP, and trace sucrose. As for non-extractives, GPs contained 25.2%-44.5% lignin, 8.04%-12.7% glucan, 4.42%-7.05% xylan, and trace amounts of galactan, arabinan, and mannan (less than 3% in total). Potential usages of these components were further examined to provide information on better valorization of GP. Considering the valuable extractives (e.g., polyphenols and oil) and non-extractives (e.g., lignin), designing a biorefinery process aiming at fully recover and/or utilize these components is of future significance.

Utilization of Lignin in Biopolymeric Packaging Films.

Lignin is a byproduct of agricultural industries and only has limited applications. In this study, lignin was investigated for use in sustainable biopolymeric packaging film. Alkali lignin (AL) and lignosulfonate (LSS) were added to enzymatically modified soy protein isolate (SPI) biopolymeric film with different concentrations with the goal of improvement of film physical and functional properties. A radical scavenging activity test revealed that films containing LSS had values 28 and 6% higher than control and AL-based films, respectively; AL itself (not in films) had significantly higher radical scavenging activity than LSS. This indicates the activity of lignin is affected by interaction with SPI. The higher compatibility between LSS and enzymatically modified SPI resulted in a positive effect on surface smoothness, water absorption, and mechanical properties of LSS-based films. Films containing AL showed a high light absorption range in the UV region, and this UV-blocking ability increased with increasing level of lignin. Deconvoluted Fourier transform infrared spectra confirmed that the addition of lignin resulted in some changes in the secondary structure of the protein matrix, which were aligned with X-ray diffraction results. The addition of lignin improved tensile strength (TS) and thermal stability of films compared to the lignin-free control. This improvement in TS and thermal stability was probably a result of new intermolecular interactions between lignin and SPI. Water vapor permeability of the films containing lignin decreased to 50% of the control because lignin played a role as a filler in the matrix. On the basis of our observations, the incorporation of lignin into biopolymeric film is capable of providing additional benefits and solutions to various industries, such as food, packaging, agriculture, and pharmaceuticals.

Evaluation of Enzymatically Modified Soy Protein Isolate Film Forming Solution and Film at Different Manufacturing Conditions.

The effects of transglutaminase on soy protein isolate (SPI) film forming solution and films were investigated by rheological behavior and physicochemical properties based on different manufacturing conditions (enzyme treatments, enzyme incubation times, and protein denaturation temperatures). Enzymatic crosslinking reaction and changes in molecular weight distribution were confirmed by viscosity measurement and SDS-PAGE, respectively, compared to 2 controls: the nonenzyme treated and the deactivated enzyme treated. Films treated with both the enzyme and the deactivated enzyme showed significant increase in tensile strength (TS), percent elongation (%E), and initial contact angle of films compared to the nonenzyme control film due to the bulk stabilizers in the commercial enzyme. Water absorption property, protein solubility, Fourier transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy revealed that enzyme treated SPI film matrix in the molecular structure level, resulted in the changes in physicochemical properties. Based on our observation, the enzymatic treatment at appropriate conditions is a practical and feasible way to control the physical properties of protein based biopolymeric film for many different scientific and industrial areas. PRACTICAL APPLICATION: Enzymes can make bridges selectively among different amino acids in the structure of protein matrix. Therefore, protein network is changed after enzyme treatment. The behavior of biopolymeric materials is dependent on the network structure to be suitable in different applications such as bioplastics applied in food and pharmaceutical products. In the current research, transglutaminase, as an enzyme, applied in soy protein matrix in different types of forms, activated and deactivated, and different preparation conditions to investigate its effects on different properties of the new bioplastic film.

New approach for characterization of gelatin biopolymer films using proton behavior determined by low field 1H NMR spectrometry.

The behavior of protons in biopolymer films (BFs) formed with gelatin, water, and glycerol was investigated at various relative humidities (RHs) and concentrations of glycerol using a low field 1H NMR spectrometer. At a RH of approximately 0%, the distributed spin-spin relaxation times (T2) of protons in BFs showed two components: a rapidly relaxing proton with the shortest T2 derived from protons in the rigid backbone of the gelatin polymer such as CH1-, CH2-, and CH3-, and a slowly relaxing component with longer T2 from protons of the functional groups in amino acid residues in gelatin such as -OH, -COOH, and -NH3. These two components are referred to as nonexchangeable (T2N) and exchangeable protons (T2E), respectively, indicating the different mobility of the protons. The T2E increased as RH increased indicating the increase in relative mobility of protons due to the larger free volume in the BF matrix. Above a RH of 33%, the slowest relaxing component was found in all BFs and referred to as hydration-water protons (T2W) with the highest relative mobility of all protons in the films. It suggests that the free volume in BFs can be formed above a RH of 33% in the absence of glycerol. The behaviors of T2N, T2E, and T2W reveal the formation of free volume in the BF matrix associated with the presence of plasticizers (water and glycerol). The T2 behavior in BFs is consistent with the behavior of spin-lattice relaxation (T1). Our result is the first attempt to characterize using low field 1H NMR technology how all protons in a film matrix behave and to develop correlations between proton mobility and free volume in protein-based BFs plasticized with water and glycerol.

Improved thermal stability of polylactic acid (PLA) composite film via PLA-ß-cyclodextrin-inclusion complex systems.

The effects of the incorporation of PLA-ß-cyclodextrin-inclusion complex (IC) and ß-cyclodextrin (ß-CD) on biopolyester PLA films were investigated. Thermal stability, surface morphology, barrier, and mechanical properties of the films were measured at varying IC (1, 3, 5, and 7%) and ß-CD (1 and 5%) concentrations. The PLA-IC-composite films (IC-PLA-CFs) showed uniform morphological structure, while samples containing ß-CD (ß-CD-PLA-CFs) showed high agglomeration of ß-CD due to poor interfacial interaction between ß-CD and PLA moieties. According to the thermal property analysis, the 5% IC-PLA-CFs showed 6.6 times lower dimensional changes (6.5%) at the temperature range of 20-80°C than that of pure PLA film (43.0%). The increase of IC or ß-CD content in the PLA-composite films shifted the glass transition and crystallization temperature to higher temperature regions. The crystallinity of both composite films improved by increasing IC or ß-CD content. Both composite films had higher oxygen and water vapor permeability as IC or ß-CD content increased in comparison to pure PLA film. All the composite films had less flexibility and lower tensile strength than the pure PLA film. In conclusion, this study shows that the IC technique is valuable to improve the thermal expansion stability of PLA-based films.

Rapid aptasensor capable of simply diagnosing prostate cancer.

Using guanine (G)-rich DNA aptamer-conjugated 6-carboxyfluorescein (6-FAM) capable of rapidly capturing prostate specific antigen (PSA) in human serum, cost-effective and simple biosensor with guanine chemiluminescence detection was developed for early diagnosis of prostate cancer. Free G-rich DNA aptamer-conjugated 6-FAM emits bright light in guanine chemiluminescence reaction based on the principle of chemiluminescent resonance energy transfer (CRET). However, G-rich DNA aptamer-conjugated 6-FAM bound with PSA cannot emit light because PSA acts as a strong interference in CRET between 6-FAM and high-energy intermediate formed from the reaction of 3,4,5-trimethoxylphenylglyoxal (TMPG) and guanine of G-rich DNA aptamer. A chemiluminescent biosensor, developed using the different properties of G-rich DNA aptamer-conjugated 6-FAM in the absence and presence of PSA in guanine chemiluminescence reaction, was able to quantify trace levels of PSA in human serum within 30 min without time-consuming and complicated procedures (e.g., multiple incubation and washings) required for conventional immunoassays operated with expensive and intractable antibodies. The limit of detection of chemiluminescent biosensor having a wide linear dynamic range (1.9-125 ng/ml) was 1.0 ng/ml. The excellent correlation (R=0.985) between chemiluminescent biosensor and conventional enzyme immunoassay indicates that the accurate, precise, and rapid chemiluminescent biosensor can be applied as a new method for early diagnosis of prostate cancer.

Rapid and simple G-quadruplex DNA aptasensor with guanine chemiluminescence detection.

Cost-effective and sensitive aptasensor with guanine chemiluminescence detection capable of simply quantifying thrombin in human serum was developed using thrombin aptamer (TBA), one of the G-quadruplex DNA aptamers, without expensive nanoparticles and complicated procedures. Guanines of G-quadruplex TBA-conjugated carboxyfluorescein (6-FAM) bound with thrombin do not react with 3,4,5-trimethoxylphenylglyoxal (TMPG) in the presence of tetra-n-propylammonium hydroxide (TPA), whereas guanines of free TBA- and TBA-conjugated 6-FAM immobilized on the surface of graphene oxide rapidly react with TMPG to emit light. Thus, guanine chemiluminescence in 5% human serum with thrombin was lower than that without thrombin when TBA-conjugated 6-FAM was added in two samples and incubated for 20 min. In other words, the brightness of guanine chemiluminescence was quenched due to the formation of G-quadruplex TBA-conjugated 6-FAM bound with thrombin in a sample. High-energy intermediate, capable of emitting dim light by itself, formed from the reaction between guanines of TBA and TMPG in the presence of TPA, transfers energy to 6-FAM to emit bright light based on the principle of chemiluminescence energy transfer (CRET). G-quadruplex TBA aptasensor devised using the rapid interaction between TBA-conjugated 6-FAM and thrombin quantified trace levels of thrombin without complicated procedures. The limit of detection (LOD = background + 3 × standard deviation) of G-quadruplex TBA aptasensor with good linear calibration curve, accuracy, precision, and recovery was as low as 12.3 nM in 5% human serum. Using the technology reported in this research, we expect that various types of G-quadruplex DNA aptasensors capable of specifically sensing a target molecule such as ATP, HIV, ochratoxin, potassium ions, and thrombin can be developed.

Rapid monitoring of alkaline phosphatase in raw milk using chemiluminescence detection.

A simple biosensor with 1,1'-oxalyldiimidazole chemiluminescence (ODI-CL) detection capable of rapidly quantifying and screening alkaline phosphatase (ALP) in raw and pasteurized milk was developed as an indicator for confirming whether commercial milk is properly pasteurized. Fluorescein was formed when standards containing 1.0% milk with different activities of ALP and samples containing 1.0% raw milk were incubated with fluorescein diphosphate (FDP) for 15 min at room temperature. The relative CL intensity of fluorescein measured with the addition of 80 mM H2O2 and ODI formed from the reaction of 2.0 µM bis(2,4,6-trichlorophenyl) oxalate and 10.0 µM 4-methyl imidazole in ethyl acetate was proportional to the concentration of ALP in milk. The range (39∼2500 mU/L) of linear calibration curve (R2 = 0.998) for the quantification of ALP in milk using ODI-CL detection was wider than those using currently applied fluorescence and 1,2-dioxetane CL detections. Also, the limit of detection (3.7 mU/L) determined using the former detection, which has good precision, was lower than those reported using the latter detections. In conclusion, the cost-effective and highly sensitive biosensor with ODI-CL detection can be applied to monitor whether milk is pasteurized according to acceptable ALP activities threshold level (350 mU/L) for public safety newly adopted by US and EU and the internal investigation level (100 mU/L) proposed by EU.

Evidence of multiple pathways capable of emitting peroxyoxalate chemiluminescence using a charge coupled device spectrometer.

Peroxyoxalate chemiluminescence (PO-CL) spectra obtained simultaneously and continuously using a CCD spectrometer provide evidence of the complexity of PO-CL reactions.

Microfluidic device capable of sensing ultrafast chemiluminescence.

Based on the principle of liquid core waveguide, a novel microfluidic device with micro-scale detection window capable of sensing flashlight emitted from rapid 1,1'-oxalyldi-4-methylimidazole (OD4MI) chemiluminescence (CL) reaction was fabricated. Light emitted from OD4MI CL reaction occurring in the micro-dimensional pentagonal detection window (length of each line segment: 900.0 microm, depth: 50.0 microm) of the microfluidic device with two inlets and one outlet was so bright that it was possible to take an image every 1/30 s at the optimal focusing distance (60 cm) using a commercial digital camera. Peaks obtained using a flow injection analysis (FIA) system with the micro-scale detection window and OD4MI CL detection show excellent resolution and reproducibility without any band-broadening observed in analytical devices having additional reaction channel(s) to measure light generated from slow CL reaction. Maximum height (H(max)) and area (A) of peak, reproducibility and sensitivity observed in the FIA system with the microfluidic device and OD4MI CL detection depends on (1) the mole ratio between bis(2,4,6-trichlorophenyl) oxalate and 4-methyl imidazole yielding OD4MI, (2) the flow rate to mix OD4MI, H(2)O(2) and 1-AP in the detection window of the microfluidic device, and (3) H(2)O(2) concentration. We obtained linear calibration curves with wide dynamic ranges using H(max) and A. The detection limit of 1-AP determined with H(max) and A was as low as 0.05 fmole/injection (signal/background=3.0).