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An introduction to

Phosphorescence Spectroscopy

. . . 9 Table of Phosphorescence Data . . . . . . . . . . . . . . . . . . . . . . . . 3 Theory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Sample Measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Table of Contents Table of Contents . . . . . . . . . . . . 7 Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 Sample Preparation . . . . . . . . . . . . 2 Introduction . . . 6 Sample Handling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14-15 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

From this S1 level. three possible routes can return the molecule to the ground state. The introduction of a microprocessor-based phosphorimeter using a pulsed xenon source will provide the analyst with much greater freedom and ease of operation which should stimulate greater interest in the technique. The first analytical uses of phosphorescence were published in 1957 by Kiers et al [1]. Theory The ground state of most organic compounds is the singlet state S0. The reluctance to use phosphorescence probably arises from the practical aspect of measuring the signal since cryogenic temperatures. Simplified energy level diagram of a polyatomic molecule. depending upon the wavelength (frequency) of the radiation. As well as offering selectivity through the use of excitation and emission wavelengths. are normally required. the technique has still not been widely accepted apart from a few selected areas such as pharmaceutical analysis and forensic science.Introduction Phosphorescence has been observed from a wide variety of compounds and is differentiated from fluorescence by the long-lived emission of light after extinction of the excitation source. the molecule will rapidly relax (10-12-10-14 sec) by a non-radiative route (internal conversion) to the lowest excited singlet state. Fluorescence to phosphorescence ratios can be used to aid the identification of particular compounds and the purity of organic compounds can be determined by measuring their decay times. c o m 3 . S1. Background interference from fluorescence and Rayleigh and Raman scattering can be rejected on a time basis. and the absorption of light promotes the electrons within the molecule to any one of several excited states S1. w w w. S2. The sensitivity of phosphorescence is comparable to that of fluorescence and complements the latter technique by offering a wider range of molecules for study. However. Developments in room temperature phosphorescence (RTP) have given rise to practical and fundamental advances which should help stimulate interest in phosphorimetry. Sn. This absorption process takes about 10-15 sec to occur and while in this excited state. Figure 1. phosphorescence adds another dimension – that of time. p e r k i n e l m e r. using liquid nitrogen at 77 K.

there have been a number of other fundamental and practical advances which have increased the interest in room temperature phosphorescence (RTP).A non-radiative internal conversion (10-5 to 10-7 sec) can take place with the excess energy released as heat. has been removed by heating the block at 100 ˚C for 12 hours (Figure 2). silica and other chromatographic supports. At these low temperatures. molecular collisions are greatly reduced and strong phosphorescence signals are observed. and thorough degassing of the solution is required before measurement. Since their publication. Several methods have been used to enable the observation of phosphorescence. bimolecular collision and photodecompositions. Phosphorescence emission spectra occur at longer wavelengths than fluorescence emission spectra because of the slight loss in energy which occurs in going from the singlet to triplet state. Quenching of the triplet state by oxygen is also effective in preventing phosphorescence. the most important feature is the addition of heavy atoms which improves the sensitivity of the technique. Because of the long lifetimes. The latter has a long lifetime which can vary from 10-6 to 102 sec depending upon the structure of the molecule. Among these advances. usually at the temperature of liquid nitrogen (77 K). although the area of applications is limited. Developments have shown that. One of the most common techniques is to supercool solutions to a rigid glass state. the molecule has a very high probability of losing its excess energy by radiativeless routes such as internal conversion. 4 . Fluorescence can also be observed at low temperatures with a sharpening of the emission peaks through the Shpol’skii effect. Figure 2. [2] In molecules where the singlet S1 and triplet T1 energy levels are closely spaced. to restrict collisional deactivation. In 1972. phosphorescence is not routinely observed in solutions at room temperature. under certain circumstances. in other words. the molecule can drop into the lower energy T1 state through a process known as intersystem crossing (10-8 sec). which may have diffused into the block. Phosphorescence can also be observed by inserting the analyte into a rigid polymer matrix. As a result. Phosphorescence emission spectrum of coronene in PMMA. The molecule can then return to the ground state by emission of radiation and this T1 to S1 transition is called phosphorescence. The phosphorescence of coronene embedded in a polymethylmethacrylate block can be observed but only after oxygen. Schulman and Walling [3] published two papers indicating that phosphorescence of sufficient strength for analytical purposes could be observed from polar organic molecules adsorbed onto filter paper. the phosphorescence of polyatomic aromatic compounds adsorbed on a variety of supports can be observed at room temperature. A radiative transition can occur (10-6 to 10-9 sec) to the various vibrational states that form the ground state S0 and this is called fluorescence.

inorganic salts and oxides such as the rare earths. c o m 5 . called EPA. which would lead to poor solidification and cracking of the glass. phosphorescence can also be observed from solid samples at room temperature. Stray light from the excitation monochromator is rejected in the phosphorometric mode.The added dimension of time imparts a number of advantages for the analyst. Figure 3 shows the phosphorescence spectrum of salicylic acid adsorbed onto a paper surface from a solution containing 1 N sodium hydroxide and 1 N sodium iodide. The samples are placed in a narrow quartz tube (internal diameter varying from 1 to 3 mm). The sample tubes are placed in liquid nitrogen held in a quartz Dewar flask. Sample preparation The majority of phosphorescence measurements are carried out in rigid media at the temperature of liquid nitrogen. europium and uranium. lead. Raman bands no longer interfere with the analysis since the Raman phenomenon has essentially the same time scale as fluorescence. The criteria for solvent selection are: · good solubility of the analyte · formation of a clear rigid glass at 77 K · low phosphorescence background (high purity) Unfortunately. silver. for example iodine. relatively few solvents form clear solids at 77 K. For non-polar compounds. w w w. and too large a diameter. p e r k i n e l m e r. Other solvents used are hexane. Polar organic molecules. cellulose. isopentane and ethanol in the ratio of 5:5:2 respectively. Phosphorescence is observed from these compounds by holding them in a powder holder of the solid sample accessory fitted to the PerkinElmer Model LS-55. As mentioned earlier. it is a nanosecond event. and clearly the selection of the right solvent plays an important part in obtaining accurate measurements. silica. when absorbed onto filter paper from solutions containing 1 N NaOH and thoroughly dried. ethanol is an excellent solvent and small quantities of base or acid may be added to produce a clear solid. which would give poor signal response and high scatter. isopentane and heptane. The dimensions are a compromise between too small a diameter. and the compounds can be divided into two types. Further studies have shown that the phosphorescence could be enhanced by the addition of heavy atoms. though even the spectrograde quality requires distillation to reduce the background phosphorescence signal. In addition. in other words. as is second order scattering. For polar compounds. exhibit phosphorescence. and the latter placed in the sample holder known as a phosphoroscope. Room temperature phosphorescence of salicylic acid adsorbed onto paper. the most popular solvent is a mixture of diethyl ether. etc. The second type of compounds are those which exhibit phosphorescence when absorbed onto certain substrates such as paper. which phosphoresce naturally without any sample pre-treatment. The first includes Figure 3.

Sample measurement Two types of phosphoroscopes are used to measure phosphorescence. Radiation from the excitation monochromator passes onto the rotating can and. the excitation beam is blocked and the fluorescence and scattered radiation decay to a negligible amount. radiation falls on the sample and excites both fluorescence and phosphorescence. · A pulsed source produces higher peak intensities than a continuously operated xenon lamp resulting in a greater peak phosphorescence emission intensity.1 to 50 msec) [7]. Figure 4 shows the excitation and emission spectra of a number of halogenated biphenyls at 77 K. · Pulsed source phosphorimetry has the advantage of time resolution compared with a mechanically modulated system permitting the analysis of organic phosphors with short lifetimes (0. Further rotation of the can will bring the window into alignment with the emission monochromator entrance slit and phosphorescence radiation will pass into the emission monochromator and onto the sample photomultiplier. Pulsed source-time resolved phosphorimetry was first proposed by Fisher and Winefordner in 1972 [6] and with the introduction of the PerkinElmer Model LS-5 luminescence spectrometer. During the cycle. The first type is known as the rotating disk phosphoroscope. the advantages of this system compared with the conventional mechanical chopper are realized. The emission monochromator is blocked by the can so that fluorescence and scattered radiation cannot be detected. with three open and three larger opaque areas. These advantages are as follows: 6 Figure 4. This minimizes variation in the signal due to sample inhomogeniety resulting from imperfect glass formation at low temperatures [4. Porro. when the window in the can is aligned with the excitation beam. aqueous solutions which have not frozen correctly. a recorder trace of the decay with respect to time can be produced. Phosphorescence spectra have been observed from “snows”. the phosphorescence intensity increases to a peak value (I0) and then theoretically decays exponentially (in practice. Courtesy T. Low temperature phosphorescence spectra of halogenated biphenyls. Figure 5 shows the schematic diagram of the events occurring during the excitation of a sample with the pulsed xenon source in the phosphorescence mode. The rotating can or chopper rotates at speeds greater than 1000 rpm. The signals from the sample photomultiplier are gated and both the delay of the start of the gate after the start of the flash (td). By measuring the phosphorescence intensity at several time intervals along the emission decay curve. A rotating disk excitation optical chopper.J. Timing is performed by a crystal clock and is highly accurate. As the can rotates. The analytical precision and accuracy for quantitative measurements is improved by rotating the sample tube. in other words. is used to alternately excite the sample and allow phosphorescence to be measured. . using this technique. 5]. The xenon lamp produces a burst of energy with a width at half peak intensity of less than 10 µsec. The second type is the rotating can phosphoroscope. During this period. and the duration of the gate (tg) can be varied in multiples of 10 µsec. the shape of the decay curve may differ due to a number of factors such as solvent interaction and the presence of more than one compound). a quantum-corrected reference photomultiplier is used to monitor the flash intensity and the signals from the sample and reference photomultiplier are ratioed to compensate for any source instability and to provide corrected excitation spectra.

Plot of log intensity against time in msec for 0. c o m 7 . dissolving in a suitable solvent and measuring the phosphorescence at 77 K [9].54% Eu in Y2O3. The technique can also be applied to room temperature phosphorescence [11]. The added dimension of time can be used to discriminate between background fluorescence and Rayleigh and Raman scatter. work has shown that phosphorimetry can be successfully combined with HPLC and TLC. p e r k i n e l m e r. the scatter is not observed and the true emission spectrum of the uranyl ion is seen. Figure 5. The earlier studies involved scraping the sample from the TLC plate. Lloyd [8] described packing a flow cell with a dry mixture of lint and crushed quartz to determine various enzyme compounds in an aqueous free solvent.54 mol%. The lifetime of the various transitions can be easily measured by monitoring the intensity with varying delay times (td) and calculating the slope of the line Figure 7. When measured in the phosphorescence mode. Figure 7 shows the results of the two such plots taken at different wavelengths. In the fluorescence mode. The method has been successfully applied to the measurement of phenothiazines. and phenytoin and to analyze 6-mercaptopurine and its metabolites in blood serum. Figure 6 shows the emission spectrum of a Eu:Y2O2 mixture measured using a solid sampling accessory. In 1975. Giffard showed that phosphorescence could be measured from a flexible TLC plate at 77 K in a modified phosphoroscope [10]. an emission spectrum containing the Rayleigh and Raman scatter superimposed upon the emission peaks of the uranyl phosphate is shown in Figure 8. Phosphorescence spectrum of Eu/Y2O3 0. obtained by plotting log intensity against time.Figure 6. w w w. Schematic diagram of events occurring during the excitation of a sample with a pulsed xenon source in phosphorescence mode. Sample handling Although the vast majority of phosphorescence measurements are made with phosphoroscopes and a solid sample holder. thiobarbiturates.

A review on the use of phosphor coated stamps and envelopes has appeared which gives a fascinating insight into the practical use of phosphorescence [20]. Europium decays with a lifetime of 0. Phosphorescence has been used in the detection of air and water-borne pollutants [15]. cocaine. Figure 8. Dinh [18] using a continuous filter paper device. Figure 9 shows two emission spectra of a 50:50 mixture of europium and terbium chloride taken at different delay times. phosphorescence mode. the reader is referred to the excellent reviews which appear annually in Analytical Chemistry. He also suggested using heavy atoms to aid the analysis of complex mixtures by RTP. particularly europium and terbium. The phosphorescence intensity of the rare earths increases tremendously when they are covalently bound to certain molecules and this feature has been used in the analysis of transferin in blood [21].74 msec. The rare earths and uranyl elements phosphoresce and a number of them. Table 1 lists the phosphorescence data taken from the literature for a number of organic compounds measured at 77 K and at room temperature. Discrimination between phosphorescence and Rayleigh and Raman scatter. An online analysis by RTP was proposed by Vo. as did Meyers for the determination of tryptophan and tyrosine [19]. are used as phosphors in lamps and TV tubes. chlorpromazine and salicylic acid. For further applications and references. Applications The majority of phosphorescence applications have been applied in the drug and pharmaceutical field [12] and in the analysis of pesticides [13]. procaine.Figure 9. Emission spectrum Tb Cl3/EuCl3 50/50 mixture. Winefordner and Tin [14] have published a paper on the determination of a number of drugs in urine and blood. Setting a short delay and sampling or gating time will have the opposite effect and favor the quickly decaying europium. A number of the sulphonamide class of drugs exhibit phosphorescence as do phenobarbital. Time-resolved spectroscopy offers an ideal way of removing unwanted background fluorescence with a consequent increase in sensitivity [22. Setting the delay to 1 msec gives an emission spectrum which is predominantly that of terbium. inhibiting the terbium from accumulating a significant signal. 23].25 msec whereas terbium has a lifetime of 0. 8 . for the analysis of impurities in polycyclic aromatic hydrocarbons [16] and in petroleum products [17]. Time can also be used to discriminate between two phosphors each with differing lifetimes.

2.10 0.2 24 25 26 27 28 28 24 29 30 14 24 27 24 14 24 3.05 0.3 2.2 0. Table 1.1 0. WE = water.0 0. WM = water. particularly in clinical chemistry [46] and the forensic. environmental and pharmaceutical fields. the introduction of new instrumentation and advances made in room temperature phosphorescence have lead to an increase in its use.3 5.001 0. NOTE: Ex (excitation) and Em (emission) wavelengths are given in nm for maximum excitation and emission.004 0.4-Benzpyrene Biphenyl 6-Bromopurine Brucine Caffeine Carbazole Chloroneb p-Chlorophenol w w w.2 0.002 0. c o m 9 . methanol in the ratio 9:1. The abbreviation RTP indicates room temperature phosphorescence carried out on various support materials.2 0.001 0.4 0.1 2.4 1.Conclusion Although the applications of phosphorescence have been somewhat limited in the past.10 0.8 0.1 1.009 < 0. Phosphorescence data for organic compounds. ethanol in the ratio 5:5:2.9 – 0. µg/ml. EPA = diethylether. isopentane.Benzanthracene Benzimidazole Benzocaine Benzoic acid 3. but in the case of RTP measurements.02 4.9 2.5 0.2 2. 1 2 Acenaphthene Adenine EtOH WM RTP EtOH EtOH WM EtOH EtOH EPA EtOH EtOH EtOH EtOH EPA EtOH EtOH WM EtOH EtOH EtOH WE EtOH 300 278 20 280 305 321 300 320 240 360 310 280 310 240 325 270 273 305 285 341 303 290 515 406 470 422 425 456 462 470 380 410 510 406 430 400 508 385 420 435 440 436 487 505 – 2.001 0. the detection limit is in ng total weight. Detection limits are quoted in concentration units. ethanol in the ratio 9:1.005 3.66 3.4 2.0 7. Various solvents are used and the abbreviations are EtOH = ethanol.007 0.1 3.2 0.006 0.0 0. p e r k i n e l m e r.8 0. The following table gives phosphorescence data for some organic compounds.0002 0.03 0. Compound Solvent Wavelengths EX EM Lifetime in sec Limit of Detection µg/ml Ref.1 25 29 14 31 32 13 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 Adenosine p-Aminobenzoic acid 6-Amino-6-methyl mercaptopurine Anthracene Apomorphine HCl Aspirin Atropine 1.

007 0.4-Dichlorophenoxyacetic acid Dicumarol 3.5-Dimethoxy-4methylamphetamine N.004 0.3 0.001 0.2 0.3 0.001 0.2 1.004 0.0 0.7 < 0.2 0.6 0.9 0.1 5.07 2.6 0.64 0.04 0.002 0.01 0.9 6. 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 o-Chlorophenoxyacetic acid p-Chlorophenoxyacetic acid 6-Chloropurine Chlorpromazine HCl Chlortetracycline Cocaine HCl Codeine Co-Ral DDT Diacetylsulphanamide 1.5 0.9 3.2.01 0. Compound Solvent Wavelengths EX EM Lifetime in sec Limit of Detection µg/ml Ref.6-Dibenzanthracene Dichloran Dichlorophen 2.6 < 0.2 0.04 0.5 0.1 0.4-Dihydroxymandelic acid 3.03 0.3 1.02 0.7 2.3 < 0.4-Dihydroxyphenylacetic acid 2.2 0.002 0.9 0.15 0.5 0.01 0.004 0.09 0.3 0.001 0.001 33 33 25 34 14 14 14 36 29 13 13 35 24 32 32 32 33 36 37 37 38 38 37 37 14 37 27 33 28 32 10 .015 0.6 2.9 3.5.05 0.Table 1 continued.003 0.0 3.2 0.028 < 0.7 0.N-Dimethyl-tryptamine Dopa Dopamine Ephedrine Epinephrine Estradiol Ethyl-3-indoleacetate Folic acid Folpet EtOH EtOH WM RTP EtOH EtOH EtOH RTP EtOH EtOH EtOH EtOH EtOH WE WE RTP EtOH EtOH EtOH EtOH WM WM EtOH EtOH EtOH EtOH EtOH EtOH EtOH WE 280 283 273 290 320 280 240 285 270 335 270 280 340 322 290 309 289 305 260 295 289 286 286 273 225 260 292 290 367 305 518 396 419 460 490 410 400 460 505 510 420 405 550 504 487 489 490 475 397 430 411 434 435 410 390 412 403 440 425 440 0.

15 0.0004 0.1 0.2 1.8 1.8 2.06 0. 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 Fluoranthene Guthion Homovanillic acid 5-Hydroxy indole acetic acid 5-Hydroxy tryptophan Ibocaine HCl Indole-3-acetic acid Kelthane Lidocaine Lysergic acid diethylamide (LSD) Metanephrine Methoxychlor 6-Methylmercaptopurine 6-Methytpurine Morphine Morphine sulphate Nalidixic acid Naphthalene -Naphthalene acetic acid -Naphthalene sulphonic acid -Naphthol -Naphtho xyacetic acid Niacinamide Nicotine 4-Nitrophenol Norepinephrine Normetanephrine Papavarine HCl Parathion RTP EtOH EtOH RTP RTP WM EtOH EtOH EtOH WM EtOH EtOH WM RTP WM RTP EtOH EtOH WM RTP EPA EtOH RTP EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH 365 325 292 312 320 292 290 285 265 311 288 275 291 290 272 268 285 265 336 338 310 295 286 320 328 270 270 355 284 290 260 360 545 420 418 510 500 430 438 515 400 512 418 380 420 458 405 449 500 460 472 438 475 510 511 475 497 410 390 520 386 416 480 515 0.2 0.004 1.8 0.7 0.01 5 0.04 0.00002 0.5 < 0.0002 0.02 0.2 < 0.5 < 0.008 0.6 1.02 0.2 0.6 3.83 1.5 0. p e r k i n e l m e r.0004 0.1 8.2 0.006 0.02 0.3 0.006 0.0006 1.05 0.01 0.01 0.3 0.15 2.2 0.2 0.7 0.0 12 0.06 0. c o m 11 .5 0.2 0.Table 1 continued.008 39 13 37 32 32 38 33 13 14 38 37 33 25 26 25 40 30 30 11 11 41 33 42 13 33 28 13 13 37 37 30 13 w w w.6 5.01 10.65 1.0005 0. Compound Solvent Wavelengths EX EM Lifetime in sec Limit of Detection µg/ml Ref.5 0.6 < 0.2 0.25 0.

0 0.7 50 0.2 1.3 1.8 2.4 1.8 0.012 3 0.06 0.1 1.01 0.0006 0.0 0.0001 0.9 3.0 1.07 0.0 0.2 2.9 1.01 1.6-Trichlorophenol EPA EPA EtOH EtOH EtOH WM RTP EtOH EtOH EtOH EtOH EtOH RTP EtOH EtOH RTP EtOH EtOH EtOH EtOH EtOH EtOH RTP EtOH EtOH EtOH RTP EtOH EtOH EtOH EtOH WE WE 360 340 240 305 310 272 330 310 291 343 340 340 320 300 315 320 300 315 290 305 280 267 280 280 297 267 310 310 312 296 305 297 410 465 380 405 430 405 600 440 425 480 500 500 510 475 430 470 410 510 410 440 405 410 426 405 410 411 426 440 420 432 430 480 482 2.4 2.2 1.05 0.9 0.6 1.2 6.7 0.0001 1.004 1.04 0.4.4.3 0.7 1.9 < 0.3 0.1 1.3 0.5-Trichlorophenol 2.05 0.Table 1 continued.04 25 0. 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 705 106 107 108 Phenacetin Phenanthrene Phenobarbital Phthalylsulphathiazole Procaine HCl Purine Pyrene Pyridine Pyridoxine HCl Quercetin Quinidine sulphate Quinine Quinoline Ronnel Salicyclic acid Serotonin Sevin Sodium sulphathiazole Strychnine phosphate Sulphabenzamide Sulphacetamide Sulphaguanidine Sulphamerazine Sulphamethazine Sulphanilamide Sulphapyridine Sulphathiazole 2-Thiouracil -Tocopherol 2.16 31 43 14 35 14 25 26 35 28 27 14 14 25 13 35 26 44 13 35 30 45 45 40 45 45 27 26 45 45 27 28 32 32 12 . Compound Solvent Wavelengths EX EM Lifetime in sec Limit of Detection µg/ml Ref.008 0.0001 0.0001 0.3 0.0001 0.0001 0.3 < 0.05 0.0 0.0038 0.4 0.2 0.5 2.5 0.

8 7. c o m 13 .5 1.Table 1 continued.01 0.002 4 0. Compound Solvent Wavelengths EX EM Lifetime in sec Limit of Detection µg/ml Ref.9 0. p e r k i n e l m e r.1 0.01 35 26 28 40 37 29 w w w.4 0.02 0. 109 110 111 112 113 Tryptophan Tyrosine Vanillin Warfarin Yohimbine HCI EtOH RTP EtOH EtOH EtOH EtOH 295 280 253 332 305 290 440 448 394 519 460 410 1.

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001 38 38 37 37 14 37 27 33 28 32 10 .methylamphetamine 40 41 42 43 44 45 46 47 48 N.9 3.6 2.N-Dimethyl-tryptamine Dopa Dopamine Ephedrine Epinephrine Estradiol Ethyl-3-indoleacetate Folic acid Folpet WM WM EtOH EtOH EtOH EtOH EtOH EtOH EtOH WE 289 286 286 273 225 260 292 290 367 305 411 434 435 410 390 412 403 440 425 440 3.1 0.004 0.015 0.2 0.9 0.9 6.3 0.01 0.02 0.9 0.6 0.6 0.3 0.15 0.0 3.2 0.

0004 0.0006 1.8 0. 49 50 51 52 Fluoranthene Guthion Homovanillic acid 5-Hydroxy indole acetic acid RTP EtOH EtOH 365 325 292 545 420 418 0.2 0.8 0.8 0.6 < 0.006 0.01 0.7 0.2 0.06 0.3 0.1 0.5 0.7 38 37 33 25 26 25 40 30 30 11 11 41 59 60 61 62 63 64 65 66 67 Metanephrine Methoxychlor 6-Methylmercaptopurine 6-Methytpurine Morphine Morphine sulphate Nalidixic acid Naphthalene ®-Naphthalene acetic acid EtOH 295 510 2.25 0.6 1.008 0.1 0.05 0.5 < 0. Limit of Compound Solvent Wavelengths EX EM Lifetime in sec Detection µg/ml Ref.2 32 32 38 33 13 14 53 54 55 56 57 58 5-Hydroxy tryptophan Ibocaine HCl Indole-3-acetic acid Kelthane Lidocaine Lysergic acid diethylamide (LSD) WM EtOH EtOH WM RTP WM RTP EtOH EtOH WM RTP EPA 311 288 275 291 290 272 268 285 265 336 338 310 512 418 380 420 458 405 449 500 460 472 438 475 0.2 1.02 0.5 0.004 1.0 12 0.06 0.01 5 0.Table 1 continued.02 0.3 0.6 3.2 42 Norepinephrine Normetanephrin 69 70 71 ®-Naphtho Niacinamide Nicotine acid 7 Naphthol xyacetic 4-Nitrophenol e .04 39 13 37 RTP RTP WM EtOH EtOH EtOH 312 320 292 290 285 265 510 500 430 438 515 400 8.01 10.0004 33 68 ®-Naphthalene sulphonic acid RTP 72 73 74 286 7 5 7 511 6 〈7 0.83 1.

5 < 0.15 2.01 0.Papavarine HCl Parathion EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH 320 328 270 270 355 284 290 260 360 475 497 410 390 520 386 416 480 515 1.2 0.02 0.0002 0.2 < 0.0005 0.6 5.00002 0.15 0. c o m 11 .006 0.2 0.2 0.008 13 33 28 13 13 37 37 30 13 w w w. p e r k i n e l m e r.65 1.5 0.

9 0.9 1.0 0.3 < 0.0006 0. Limit of Compound Solvent Wavelengths EX EM Lifetime in sec Detection µg/ml Ref.2 1.1 1.5 0.2 6.4 1.2 2.2 0.7 1.4 0.05 0.8 2.9 < 0.05 0.1 1.5 2.3 0.0 0.7 50 0.0001 0.012 3 0.3 0. 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 Phenacetin Phenanthrene Phenobarbital Phthalylsulphathiazole Procaine HCl Purine Pyrene Pyridine Pyridoxine HCl Quercetin Quinidine sulphate Quinine Quinoline Ronnel Salicyclic acid Serotonin Sevin Sodium sulphathiazole Strychnine phosphate Sulphabenzamide Sulphacetamide Sulphaguanidine EPA EPA EtOH EtOH EtOH WM RTP EtOH EtOH EtOH EtOH EtOH RTP EtOH EtOH RTP EtOH EtOH EtOH EtOH EtOH EtOH RTP EtOH EtOH EtOH RTP EtOH EtOH EtOH EtOH WE WE 360 340 240 305 310 272 330 310 291 343 340 340 320 300 315 320 300 315 290 305 280 267 280 280 297 267 310 310 312 296 305 297 410 465 380 405 430 405 600 440 425 480 500 500 510 475 430 470 410 510 410 440 405 410 426 405 410 411 426 440 420 432 430 480 482 2.004 1.8 0.008 0.07 0.5-Trichlorophenol 108 2.4 2.0001 0.9 3.0 1.0001 0.7 0.6 1.6-Trichlorophenol .05 0.0038 0.04 25 0.16 31 43 14 35 14 25 26 35 28 27 14 14 25 13 35 26 44 13 35 30 45 45 40 45 45 27 26 45 45 27 28 32 32 100 Sulphamerazine 101 Sulphamethazine 102 Sulphanilamide 103 Sulphapyridine 104 Sulphathiazole 705 2-Thiouracil 106 〈-Tocopherol 107 2.4.01 1.2 1.0001 1.06 0.Table 1 continued.3 1.3 0.0 0.04 0.0001 0.0 0.0001 0.4.3 0.01 0.

12 .

01 35 26 28 40 37 29 w .8 7. Limit of Compound Solvent Wavelengths EX EM Lifetime in sec Detection µg/ml Ref.02 0.Table 1 continued.002 4 0. 109 Tryptophan 110 Tyrosine 111 Vanillin 112 Warfarin 113 Yohimbine HCI EtOH RTP EtOH EtOH EtOH EtOH 295 280 253 332 305 290 440 448 394 519 460 410 1.4 0.1 0.5 1.9 0.01 0.

13 .

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