Development of high-throughput ATR-FTIR technology for rapid triage of brain cancer.

Non-specific symptoms, as well as the lack of a cost-effective test to triage patients in primary care, has resulted in increased time-to-diagnosis and a poor prognosis for brain cancer patients. A rapid, cost-effective, triage test could significantly improve this patient pathway. A blood test using attenuated total reflection (ATR)-Fourier transform infrared (FTIR) spectroscopy for the detection of brain cancer, alongside machine learning technology, is advancing towards clinical translation. However, whilst the methodology is simple and does not require extensive sample preparation, the throughput of such an approach is limited. Here we describe the development of instrumentation for the analysis of serum that is able to differentiate cancer and control patients at a sensitivity and specificity of 93.2% and 92.8%. Furthermore, preliminary data from the first prospective clinical validation study of its kind are presented, demonstrating how this innovative technology can triage patients and allow rapid access to imaging.

Rapid quantification of clove (Syzygium aromaticum) and spearmint (Mentha spicata) essential oils encapsulated in a complex organic matrix using an ATR-FTIR spectroscopic method.

Essential oils (EOs) are often encapsulated in various and complex matrices to protect them against potential degradation or to control their release. To achieve an optimum use in food products, their rapid and precise quantification after encapsulation and storage is required. Hence, a rapid ATR-FTIR method was developed and tested with two encapsulated essential oils (EOs): clove (Syzygium aromaticum) and spearmint (Mentha spicata);. Despite, the complexity of the matrix, this method coupled with univariate or multivariate regression models exhibited high potential for global quantification of the two encapsulated EOs. For clove EO, in relation to the major presence of eugenol and eugenol acetate, an analysis based on a unique band (1514 cm-1) was sufficient to obtain a good prediction with RMSEP value of 0.0173 g of EO per g of matrix. For spearmint oil which is characterized by numerous terpenoid compound, three bands (799, 885, and 1680-1676 cm-1) were suitable for a good prediction with RMSEP value of 0.0133. ATR-FTIR method was compared with a reference gas chromatography FID quantitative method in an EO release experiment and its efficiency was evaluated through modeling by the Avrami equation. Beside time saving, the ATR-FTIR method was also capable of monitoring the EO profile. This method could be easily adapted as a routine analysis in the EOs industry as quality control.

Detection of mycoplasma in contaminated mammalian cell culture using FTIR microspectroscopy.

Mycoplasma contamination represents a significant problem to the culture of mammalian cells used for research as it can cause disastrous effects on eukaryotic cells by altering cellular parameters leading to unreliable experimental results. Mycoplasma cells are very small bacteria therefore they cannot be detected by visual inspection using a visible light microscope and, thus, can remain unnoticed in the cell cultures for long periods. The detection techniques used nowadays to reveal mycoplasma contamination are time consuming and expensive with each having significant drawbacks. The ideal detection should be simple to perform with minimal preparation time, rapid, inexpensive, and sensitive. To our knowledge, for the first time, we employed Fourier transform infrared (FTIR) microspectroscopy to investigate whether we can differentiate between control cells and the same cells which have been infected with mycoplasmas during the culturing process. Chemometric methods such as HCA and PCA were used for the data analysis in order to detect spectral differences between control and intentionally infected cells, and spectral markers were revealed even at low contamination level. The preliminary results showed that FTIR has the potential to be used in the future as a reliable complementary detection technique for mycoplasma-infected cells. Graphical abstract FTIR microspectroscopy is able to differentiate between mycoplasma infected cells (LC for low contamination and HC for high contamination) and control non-infected cells (CN).

Infrared vibrational spectroscopy: a rapid and novel diagnostic and monitoring tool for cystinuria.

Cystinuria is the commonest inherited cause of nephrolithiasis (~1% in adults; ~6% in children) and is the result of impaired cystine reabsorption in the renal proximal tubule. Cystine is poorly soluble in urine with a solubility of ~1 mM and can readily form microcrystals that lead to cystine stone formation, especially at low urine pH. Diagnosis of cystinuria is made typically by ion-exchange chromatography (IEC) detection and quantitation, which is slow, laboursome and costly. More rapid and frequent monitoring of urinary cystine concentration would significantly improve the diagnosis and clinical management of cystinuria. We used attenuated total reflection - Fourier transform infrared spectroscopy (ATR-FTIR) to detect and quantitate insoluble cystine in 22 cystinuric and 5 healthy control urine samples. Creatinine concentration was also determined by ATR-FTIR to adjust for urinary concentration/dilution. Urine was centrifuged, the insoluble fraction re-suspended in 5 µL water and dried on the ATR prism. Cystine was quantitated using its 1296 cm-1 absorption band and levels matched with parallel measurements made using IEC. ATR-FTIR afforded a rapid and inexpensive method of detecting and quantitating insoluble urinary cystine. This proof-of-concept study provides a basis for developing a high-throughput, cost-effective diagnostic method for cystinuria, and for point-of-care clinical monitoring.

FTIR assay method for UV inactive drug carisoprodol and identification of degradants by RP-HPLC and ESI-MS.

A new method of analysis has been developed for UV inactive drug carisoprodol using FTIR spectroscopy. These methods were validated for various parameters according to ICH guidelines. The proposed method has also been successfully applied for the determination of the drug concentration in a tablet formulation. The method proved to be accurate (mean percentage recovery between 95 and 105%), precise and reproducible (relative standard deviation<2%), while being simple, economical and less time consuming than other methods and can be used for routine estimation of carisoprodol in the pharmaceutical industry. The developed method also implicates its utility for other UV inactive substances. The stability of the drug under various stress conditions was studied and the drug was found to be particularly susceptible to alkaline hydrolysis. Degradation products of the alkaline hydrolysis were detected by RP-HPLC and tentatively identified by ESI-MS.

Rapid determination and chemical change tracking of benzoyl peroxide in wheat flour by multi-step IR macro-fingerprinting.

BPO is often added to wheat flour as flour improver, but its excessive use and edibility are receiving increasing concern. A multi-step IR macro-fingerprinting was employed to identify BPO in wheat flour and unveil its changes during storage. BPO contained in wheat flour (<3.0 mg/kg) was difficult to be identified by infrared spectra with correlation coefficients between wheat flour and wheat flour samples contained BPO all close to 0.98. By applying second derivative spectroscopy, obvious differences among wheat flour and wheat flour contained BPO before and after storage in the range of 1500-1400 cm(-1) were disclosed. The peak of 1450 cm(-1) which belonged to BPO was blue shifted to 1453 cm(-1) (1455) which belonged to benzoic acid after one week of storage, indicating that BPO changed into benzoic acid after storage. Moreover, when using two-dimensional correlation infrared spectroscopy (2DCOS-IR) to track changes of BPO in wheat flour (0.05 mg/g) within one week, intensities of auto-peaks at 1781 cm(-1) and 669 cm(-1) which belonged to BPO and benzoic acid, respectively, were changing inversely, indicating that BPO was decomposed into benzoic acid. Moreover, another autopeak at 1767 cm(-1) which does not belong to benzoic acid was also rising simultaneously. By heating perturbation treatment of BPO in wheat flour based on 2DCOS-IR and spectral subtraction analysis, it was found that BPO in wheat flour not only decomposed into benzoic acid and benzoate, but also produced other deleterious substances, e.g., benzene. This study offers a promising method with minimum pretreatment and time-saving to identify BPO in wheat flour and its chemical products during storage in a holistic manner.

Fast IR laser mapping ellipsometry for the study of functional organic thin films.

Fast infrared mapping with sub-millimeter lateral resolution as well as time-resolved infrared studies of kinetic processes of functional organic thin films require a new generation of infrared ellipsometers. We present a novel laboratory-based infrared (IR) laser mapping ellipsometer, in which a laser is coupled to a variable-angle rotating analyzer ellipsometer. Compared to conventional Fourier-transform infrared (FT-IR) ellipsometers, the IR laser ellipsometer provides ten- to hundredfold shorter measurement times down to 80 ms per measured spot, as well as about tenfold increased lateral resolution of 120 µm, thus enabling mapping of small sample areas with thin-film sensitivity. The ellipsometer, equipped with a HeNe laser emitting at about 2949 cm(-1), was applied for the optical characterization of inhomogeneous poly(3-hexylthiophene) [P3HT] and poly(N-isopropylacrylamide) [PNIPAAm] organic thin films used for opto-electronics and bioapplications. With the constant development of tunable IR laser sources, laser-based infrared ellipsometry is a promising technique for fast in-depth mapping characterization of thin films and blends.

Large scale infrared imaging of tissue micro arrays (TMAs) using a tunable Quantum Cascade Laser (QCL) based microscope.

Chemical imaging in the field of vibrational spectroscopy is developing into a promising tool to complement digital histopathology. Applications include screening of biopsy tissue via automated recognition of tissue/cell type and disease state based on the chemical information from the spectrum. For integration into clinical practice, data acquisition needs to be speeded up to implement a rack based system where specimens are rapidly imaged to compete with current visible scanners where 100's of slides can be scanned overnight. Current Fourier transform infrared (FTIR) imaging with focal plane array (FPA) detectors are currently the state-of-the-art instrumentation for infrared absorption chemical imaging, however recent development in broadly tunable lasers in the mid-IR range is considered the most promising potential candidate for next generation microscopes. In this paper we test a prototype quantum cascade laser (QCL) based spectral imaging microscope with a focus on discrete frequency chemical imaging. We demonstrate how a protein chemical image of the amide I band (1655 cm(-1)) of a 2 × 2.4 cm(2) breast tissue microarray (TMA) containing over 200 cores can be measured in 9 min. This result indicates that applications requiring chemical images from a few key wavelengths would be ideally served by laser-based microscopes.

The contribution of Raman spectroscopy to the analytical quality control of cytotoxic drugs in a hospital environment: eliminating the exposure risks for staff members and their work environment.

The purpose of the study was to perform a comparative analysis of the technical performance, respective costs and environmental effect of two invasive analytical methods (HPLC and UV/visible-FTIR) as compared to a new non-invasive analytical technique (Raman spectroscopy). Three pharmacotherapeutic models were used to compare the analytical performances of the three analytical techniques. Statistical inter-method correlation analysis was performed using non-parametric correlation rank tests. The study's economic component combined calculations relative to the depreciation of the equipment and the estimated cost of an AQC unit of work. In any case, analytical validation parameters of the three techniques were satisfactory, and strong correlations between the two spectroscopic techniques vs. HPLC were found. In addition, Raman spectroscopy was found to be superior as compared to the other techniques for numerous key criteria including a complete safety for operators and their occupational environment, a non-invasive procedure, no need for consumables, and a low operating cost. Finally, Raman spectroscopy appears superior for technical, economic and environmental objectives, as compared with the other invasive analytical methods.

Rapid approach to analyze biochemical variation in rat organs by ATR FTIR spectroscopy.

ATR FTIR spectra were collected from rat tissue homogenates (myocardium, brain, liver, lung, intestine, and kidney) to analyze their biochemical content. Based on the second derivative of an average spectral profile it was possible to assign bands e.g. to triglycerides and cholesterol esters, proteins, phosphate macromolecules (DNA, RNA, phospholipids, phosphorylated proteins) and others (glycogen, lactate). Peaks in the region of 1600-1700 cm(-1) related to amide I mode revealed the secondary structure of proteins. The collected spectra do not characterize morphological structure of the investigated tissues but show their different composition. The comparison of spectral information gathered from FTIR spectra of the homogenates and those obtained previously from FTIR imaging of the tissue sections implicates that the presented here approach can be successfully employed in the investigations of biochemical variation in animal tissues. Moreover, it can be used in the pharmacological and pharmacokinetic studies to correlate the overall biochemical status of the tissue with the pathological changes it has undergone.

A cost-effective method for estimating di(2-ethylhexyl)phthalate in coastal sediments.

This study describes the development of a new method for the analysis of di(2-ethylhexyl)phthalate (DEHP) using 0.1-0.3 g of sediment sample, based on matrix solid phase dispersion (MSPD) using C18 as dispersant phase (0.4 g) and acetonitrile-water as eluting solvent (3.4 mL 1:3.25, v/v). No evaporation step is required. 3 mL of extracts were processed on-line by in-tube solid phase microextraction (IT-SPME) coupled to capillary liquid chromatography (CapLC) and diode array detector (DAD). A short analytical column Zorbax SB C18 (35×0.5 mm, 5 µm) provided suitable results. FTIR-ATR was employed for characterizing sediment samples and MSPD procedure. The total analysis time was less than 20 min (MSPD takes about 10 min). The utility of the described approach has been tested by analyzing several real samples. No matrix effect was found. Achieved precision was less than 10% for DEHP estimation. Detection limits in samples were 270 and 90 µg/kg for 0.1 and 0.3 g of taken sediment, respectively.

High-throughput phenotyping of uropathogenic E. coli isolates with Fourier transform infrared spectroscopy.

Fourier transform infrared (FT-IR) spectroscopy is an established rapid whole-organism fingerprinting method that generates metabolic fingerprints from bacteria that reflect the phenotype of the microorganism under investigation. However, whilst FT-IR spectroscopy is fast (typically 10 s to 1 min per sample), the approaches for microbial sample preparation can be time consuming as plate culture or shake flasks are used for growth of the organism. We report a new approach that allows micro-cultivation of bacteria from low volumes (typically 200 µL) to be coupled with FT-IR spectroscopy. This approach is fast and easy to perform and gives equivalent data to the lengthier and more expensive shake flask cultivations (sample volume = 20 mL). With this micro-culture approach we also demonstrate high reproducibility of the metabolic fingerprints. The approach allowed separation of different isolates of Escherichia coli involved in urinary tract infection, including members of the globally disseminated ST131 clone, with respect to both genotype and resistance or otherwise to the antibiotic Ciprofloxacin.

[Raman spectroscopy applied to analytical quality control of injectable drugs: analytical evaluation and comparative economic versus HPLC and UV / visible-FTIR]. / La spectroscopie Raman appliquée au contrôle de qualité analytique des médicaments injectables: évaluation analytique et économique comparative versus CLHP et uv/visible-iRTF.

In France, central IV admixture of chemotherapy (CT) treatments at the hospital is now required by law. We have previously shown that the shaping of Therapeutic Objects (TOs) could profit from an Analytical Quality Assurance (AQA), closely linked to the batch release, for the three key parameters: identity, purity, and initial concentration of the compound of interest. In the course of recent and diversified works, we showed the technical superiority of non-intrusive Raman Spectroscopy (RS) vs. any other analytical option and, especially for both HPLC and vibrational method using a UV/visible-FTIR coupling. An interconnected qualitative and economic assessment strongly helps to enrich these relevant works. The study compares in operational situation, the performance of three analytical methods used for the AQC of TOs. We used: a) a set of evaluation criteria, b) the depreciation tables of the machinery, c) the cost of disposables, d) the weight of equipment and technical installations, e) the basic accounting unit (unit of work) and its composite costs (Euros), which vary according to the technical options, the weight of both human resources and disposables; finally, different combinations are described. So, the unit of work can take 12 different values between 1 and 5.5 Euros, and we provide various recommendations. A qualitative evaluation grid constantly places the SR technology as superior or equal to the 2 other techniques currently available. Our results demonstrated: a) the major interest of the non-intrusive AQC performed by RS, especially when it is not possible to analyze a TO with existing methods e.g. elastomeric portable pumps, and b) the high potential for this technique to be a strong contributor to the security of the medication circuit, and to fight the iatrogenic effects of drugs especially in the hospital. It also contributes to the protection of all actors in healthcare and of their working environment.

A rapid method for detection of fumonisins B1 and B2 in corn meal using Fourier transform near infrared (FT-NIR) spectroscopy implemented with integrating sphere.

Fourier transform near infrared (FT-NIR) spectroscopy is an analytical procedure generally used to detect organic compounds in food. In this work the ability to predict fumonisin B(1)+B(2) contents in corn meal using an FT-NIR spectrophotometer, equipped with an integration sphere, was assessed. A total of 143 corn meal samples were collected in Friuli Venezia Giulia Region (Italy) and used to define a 15 principal components regression model, applying partial least square regression algorithm with full cross validation as internal validation. External validation was performed to 25 unknown samples. Coefficients of correlation, root mean square error and standard error of calibration were 0.964, 0.630 and 0.632, respectively and the external validation confirmed a fair potential of the model in predicting FB(1)+FB(2) concentration. Results suggest that FT-NIR analysis is a suitable method to detect FB(1)+FB(2) in corn meal and to discriminate safe meals from those contaminated.

Rapid strain classification and taxa delimitation within the edible mushroom genus Pleurotus through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy.

Fourier transform infrared (FT-IR) spectroscopy has been successfully applied for the identification of bacteria and yeasts, but only to a limited extent for discriminating specific groups of filamentous fungi. In the frame of this study, 73 strains - from different associated hosts/substrates and geographic regions - representing 16 taxa of the edible mushroom genus Pleurotus (Basidiomycota, Agaricales) were examined through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. A binary matrix, elaborated on the basis of presence/absence of specific absorbance peaks combined with cluster analysis, demonstrated that the spectral region 1800-600 cm(-1) permitted clear delimitation of individual strains into Pleurotus species. In addition, closely related species (e.g., Pleurotus ostreatus and Pleurotus pulmonarius) or taxa of the subgenus Coremiopleurotus demonstrated high similarity in their absorbance patterns, whereas genetically distinct entities such as Pleurotus dryinus, Pleurotus djamor, and Pleurotus eryngii provided spectra with noteworthy differences. When specific regions (1800-1700, 1360-1285, 1125-1068, and 950-650 cm(-1)) were evaluated in respect to the absorbance values demonstrated by individual strains, it was evidenced that this methodology could be eventually exploited for the identification of unknown Pleurotus specimens with a stepwise process and with the aid of a dichotomous key developed for this purpose. Moreover, it was shown that the nature of original fungal material examined (mycelium, basidiomata, and basidiospores) had an effect on the outcome of such analyses, and so did the use of different mycelium growth substrates. In conclusion, application of FT-IR spectroscopy provided a fast, reliable, and cost-efficient solution for the classification of pure cultures from closely related mushroom species.

Fourier transform infrared and Raman spectroscopies for the rapid detection, enumeration, and growth interaction of the bacteria Staphylococcus aureus and Lactococcus lactis ssp. cremoris in milk.

Staphylococcus aureus is one of the main pathogenic microorganisms found in milk and dairy products and has been involved in bacterial foodborne outbreaks in the past. Current enumeration techniques for bacteria are very time-consuming, typically taking 24 h or longer, and bacterial antagonism in the form of lactic acid bacteria (LAB) may inhibit the growth of S. aureus . Therefore, the aim of this investigation was to establish the accuracy and sensitivity of rapid nondestructive metabolic fingerprinting techniques, such as Fourier transform infrared (FT-IR) spectroscopy and Raman spectroscopy (RS), in combination with multivariate analysis techniques, for the detection and enumeration of S. aureus in milk, as well as to study the growth interaction between S. aureus and Lactococcus lactis ssp. cremoris , a common LAB. The two bacterial species were investigated both in a pure monoculture and in a combined inoculated coculture after inoculation into ultraheated milk during the first 24 h of growth at 37 °C. Plating techniques were used to obtain primary reference data for viable bacteria counts. Principal component discriminant function analysis, canonical correlation analysis, partial least-squares (PLS), and kernel PLS (KPLS) multivariate statistical techniques were employed to analyze the data. FT-IR provided very reasonable quantification results both with PLS and KPLS, the latter providing marginally better predictions, with correlation coefficients in the test set (Q(2)) and training set (R(2)) varying from 0.64 to 0.76 and from 0.78 to 0.88 for different bacterial sample combinations. RS results were less encouraging with high degrees of error and poor correlation to viable bacterial counts. S. aureus growth was not inhibited by the presence of the LAB, but metabolic fingerprinting of the coculture indicated that the phenotype of this dual bacterial culture was closer to that of pure LAB cultures. In conclusion, FT-IR spectroscopy in combination with the above multivariate techniques appears to be a promising discrimination and enumeration analytical technique for the two bacterial species. In addition, it has been demonstrated that the L. cremoris metabolic effect in milk dominates that of S. aureus even though there was no growth antagonism observed.

Fast determination of thyroid stimulating hormone in human blood serum without chemical preprocessing by using infrared spectroscopy and least squares support vector machines.

The least squares support vector machines (LS-SVM) was used to model infrared spectral data for TSH hormone secreted by thyroid, which regulates the basal metabolic rate. This model was used for direct estimation of the content of TSH in blood serum samples, and the results were comparable with those obtained with the conventional analytical method based on chemoluminescence methodology. Excellent agreement was observed between the conventional method and the newly developed calibration model based in analysis of spectral data with LS-SVM. The latter has clear advantages, because it is fast and requires no reagent once the measurements were done directly in the serum by using a simple mid-infrared spectrometer in the ATR mode. An important advantage observed in this calibration method based on LS-SVM is the remarkable capacity to avoid overfitting in the model-building step, that is, the developed method is highly robust.

A spectroscopic method to estimate the binding potency of amphiphile assemblies.

A fast and convenient spectroscopic methodology to determine the water uptake capacity of amphiphile assemblies studied in multilayer films is presented. This method was developed to provide a reliable but relatively simple tool for estimating the binding potency of such complex systems. The water-binding potency represents a general propensity of higher-order systems to bind or embed relevant ligands, such as various non-lipid effectors in the case of artificial lipid membranes. In this sense, the binding potency might contribute to a specific functional role of certain lipids. The essence of the new method is that the calibration of data measured by infrared (IR) spectroscopy against those directly obtained by Karl-Fischer titration (KFT) enables one to replace the expensive chemical-analytical technique by a more comfortable and efficient IR-spectroscopic protocol. This approach combines the easy handling, versatility, and availability of IR spectroscopy with the high accuracy of KFT. The usefulness of the procedure is demonstrated on an example set of six amphiphiles with a common chain length of 18 carbon atoms. Despite this similarity, the binding potency data differ tremendously in a way which can be correlated with the systematic variations introduced into the amphiphile structure. Going further beyond the methodical aspect, the scientific relevance of the data is comprehensively discussed especially in terms of the structural factors that govern the binding potency of amphiphiles. That is favored mainly by fluidity and disfavored mainly by inter-amphiphile binding networks. For phosphatidylcholine, our data are strongly in favor of a particular hydration model that involves primary water binding to phosphate as well as the formation of water semi-clathrates hosting the trimethylammonium moiety. Interestingly, stearylamine and diolein assemblies did not take up any water at all. This unexpected hydrophobicity is due to the unusual structures formed in these latter cases: rigid ammonium amide with a strong hydrogen-bonding/salt bridge network in stearylamine, and patches of inverted micelles in diolein, as revealed by molecular dynamics simulations.

Attenuated total reflection Fourier transform infrared spectroscopy for on-chip monitoring of solute concentrations.

We report a cost-efficient Attenuated Total Reflection Fourier Transform Infrared Spectroscopy (ATR-FTIR) method for monitoring concentrations of solutes in solutions flowing through microfluidic channels. The method allows rapid acquisition of spectra and enables chemical characterisation and concentration measurements that are independent of the flow rate of liquids. The method enables independent measurement of concentrations of solutes with distinct spectral features in mixed solutions. For the polymer solutes studied in the present work, the method has a sensitivity of at least 10 microM (0.01 wt%). We also propose the applicability of the method for the differentiation between dissolved and adsorbed amphiphilic species.

Use of FT-IR to identify enhanced biomass production and biochemical pool shifts in the marine microalgae, Chlorella ovalis, cultured in media composed of different ratios of deep seawater and fermented animal wastewater.

Growth rates, photosystem II photosynthesis, and the levels of chlorophyll a and secondary metabolites of Chlorella ovalis were estimated to determine if they were enhanced by the addition of swine urine (BM) or cow compost water (EP) that had been fermented by soil bacteria to deep seawater (DSW) in an attempt to develop media that enabled batch mass culture at lower costs. Growth of C. ovalis in f/2, f/2-EDTA+BM60%, DSW+BM30%, and DSW+EP60% was enhanced and maintained in the log phase of growth for 16 days. The cell densities of C. ovalis in DSW+EP60% (4.1x10(6) Cells/ml) were higher than those of f/2 (2.9x10(6) Cells/ml), f/2-E+BM60% (3.7x10(6) Cells/ml), and DSW+BM30% (2.7x10(6) Cells/ml). The growth rate was also more favorable for C. ovalis cultured in DSW+EP60% (0.15 day(-1)) than that of C. ovalis cultured in the control medium (f/2) (0.12 day(-1)). Furthermore, the chlorophyll a concentration of C. ovalis cultured in DSW+EP60% (4.56 mg/l) was more than 2-fold greater than that of C. ovalis cultured in f/2 (2.35 mg/l). Moreover, the maximal quantum yields of photosystem II at 470 nm (Fv/Fm) were significantly higher in organisms cultured at f/2-E+BM60% (0.53) and DSW+EP60% (0.52) than in the other treatment groups. Finally, Fourier transformation infrared (FT-IR) spectroscopy revealed that C. ovalis grown in DSW+EP60% had more typical peaks and various biochemical pool shifts than those grown in other types of media. Taken together, the results of this study indicate that the use of DSW+EP60% to culture C. ovalis can reduce maintenance expenses and promote higher yields.