Pyranine (HPTS)
(Synonyms: 溶剂绿7; HPTS; Solvent Green 7) 目录号 : GC30145
A cell-impermeable fluorescent pH probe
Cas No.:6358-69-6
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Cell experiment: |
PAAm-κC composite gels are prepared by free-radical copolymerization using 0.71 g of AAm, 0.01 g of BIS (N, N’-methylenebisacrylamide), 0.008 g of APS (ammonium persulfate) and 2 µl of TEMED (tetramethylethylenediamine) are dissolved in 5 mL of distilled water (pH 6.5) by heating. The heated mixture solution is held at 80°C. Then different amounts of κ-carrageenan (0.5, 1, 1.5, 2, 2.5 and 3 (w/v) % κC) are added. Pyranine (Py) is used in the PAAm–κC composites as a fluorescence probe, which is a derivative of pyrene possessing three SO3- groups, which can form bonds with positive charges on the gel. Pyranine (Py) concentration is kept constant at 4×10-4 M, for all experiments. The solution is stirred (200 rpm) for 15 min to achieve a homogenous sample solution. All samples are deoxygenated by bubbling nitrogen for 10 min just before the polymerization process. The drying and swelling experiments of disc-shaped PAAm–κC composite gels prepared are performed in air and in water, respectively, at various temperatures (30, 40, 50, and 60°C). A model LS-50 spectrometer equipped with a temperature controller is used for fluorescence intensity measurements, which are made at a 90° position and spectral bandwidths are kept at 5 nm. Disc-shaped gel samples are placed on the wall of a 1-cm path-length square quartz cell filled with air and/or water for the drying and swelling experiments[3]. |
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References: [1]. Saha T, et al. In vitro sensing of Cu(+) through a green fluorescence rise of pyranine. Photochem Photobiol Sci. 2014 Oct;13(10):1427-33. |
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8-Hydroxypyrene-1,3,6-trisulfonic acid (HPTS) is a cell-impermeable fluorescent pH probe.1,2 It displays excitation maxima of 403 and 450 nm, the intensities of which decrease and increase, respectively, as pH increases from 5 to 9 and an emission maximum of 510 nm.1 HPTS has been used in excitation ratio imaging to monitor changes in and measure the pH of endocytosed liposomes, acidic organelles, or the cytoplasm in live cell applications.1,2 It is sensitive to fluorescence quenching by viologens, which can be reversed in the presence of glucose or other monosaccharides, and has been used as a glucose and carbohydrate sensor.3
1.Daleke, D.L., Hong, K., and Papahadjopoulos, D.Endocytosis of liposomes by macrophages: binding, acidification and leakage of liposomes monitored by a new fluorescence assayBiochim. Biophys. Acta1024(2)352-366(1990) 2.Han, J., and Burgess, K.Fluorescent indicators for intracellular pHChem. Revs.110(5)2709-2728(2010) 3.Cordes, D.B., and Singaram, B.A unique, two-component sensing system for fluorescence detection of glucose and other carbohydratesPure Appl. Chem.84(11)2183-2202(2012)
| Cas No. | 6358-69-6 | SDF | |
| 别名 | 溶剂绿7; HPTS; Solvent Green 7 | ||
| Canonical SMILES | O=S(C1=C(C2=C34)C=CC4=C(O)C=C(S(=O)([O-])=O)C3=CC=C2C(S(=O)([O-])=O)=C1)([O-])=O.[Na+].[Na+].[Na+] | ||
| 分子式 | C16H7Na3O10S3 | 分子量 | 524.39 |
| 溶解度 | Water: 100 mg/mL (190.70 mM; Need ultrasonic);DMSO: 25 mg/mL (47.67 mM; Need ultrasonic) | 储存条件 | Store at -20°C,protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.907 mL | 9.5349 mL | 19.0698 mL |
| 5 mM | 0.3814 mL | 1.907 mL | 3.814 mL |
| 10 mM | 0.1907 mL | 0.9535 mL | 1.907 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
The Dual Use of the Pyranine (HPTS) Fluorescent Probe: A Ground-State pH Indicator and an Excited-State Proton Transfer Probe
Acc Chem Res 2022 Sep 20;55(18):2728-2739.36053265 PMC9494743
Molecular fluorescent probes are an essential experimental tool in many fields, ranging from biology to chemistry and materials science, to study the localization and other environmental properties surrounding the fluorescent probe. Thousands of different molecular fluorescent probes can be grouped into different families according to their photophysical properties. This Account focuses on a unique class of fluorescent probes that distinguishes itself from all other probes. This class is termed photoacids, which are molecules exhibiting a change in their acid-base transition between the ground and excited states, resulting in a large change in their pKa values between these two states, which is thermodynamically described using the Förster cycle. While there are many different photoacids, we focus only on pyranine, which is the most used photoacid, with pKa values of ∿.4 and ∿.4 for its ground and excited states, respectively. Such a difference between the pKa values is the basis for the dual use of the pyranine fluorescent probe. Furthermore, the protonated and deprotonated states of pyranine absorb and emit at different wavelengths, making it easy to focus on a specific state. Pyranine has been used for decades as a fluorescent pH indicator for physiological pH values, which is based on its acid-base equilibrium in the ground state. While the unique excited-state proton transfer (ESPT) properties of photoacids have been explored for more than a half-century, it is only recently that photoacids and especially pyranine have been used as fluorescent probes for the local environment of the probe, especially the hydration layer surrounding it and related proton diffusion properties. Such use of photoacids is based on their capability for ESPT from the photoacid to a nearby proton acceptor, which is usually, but not necessarily, water. In this Account, we detail the photophysical properties of pyranine, distinguishing between the processes in the ground state and the ones in the excited state. We further review the different utilization of pyranine for probing different properties of the environment. Our main perspective is on the emerging use of the ESPT process for deciphering the hydration layer around the probe and other parameters related to proton diffusion taking place while the molecule is in the excited state, focusing primarily on bio-related materials. Special attention is given to how to perform the experiments and, most importantly, how to interpret their results. We also briefly discuss the breadth of possibilities in making pyranine derivatives and the use of pyranine for controlling dynamic reactions.
Supramolecular Construction of a [16]-Imidazolium Cage via a Quadruple [2+2] Photocycloaddition and Its Selective Fluorescent Recognition of Pyranine (HPTS)
Chemistry 2020 Jun 5;26(32):7190-7193.32141658 10.1002/chem.202001138
Polyimidazolium-based cages are considered promising materials based on their fascinating properties and potential applications. These three-dimensional functional structures are highly desirable for the recognition of particular guest molecules, however, their synthesis remains challenging. In this work, we have designed and synthesizes a pure [n]-imidazolium (n=16) cage, the hexadecakisimidazolium salt H16 -2(PF6 )16 , from tetragonal octakisimidazolium salt H8 -1(PF6 )8 . The synthetic method involves formation of metal-carbene templates, a quadruple photochemical [2+2] cycloaddition reaction and subsequent removal of metal ions. Specifically, the synthesized cage, featuring sixteen imidazolium moieties, demonstrated high efficiency for the selective fluorescent recognition of 8-hydroxy-1,3,6-pyrene trisulfonate (HPTS). The present work not only further develops the metal-carbene template strategy by exploiting a new type of polyimidazolium cage, but also provides encouraging prospects for the design of versatile imidazolium-based functional acceptors.
Deprotonation dynamics and stokes shift of Pyranine (HPTS)
J Phys Chem A 2007 Jan 18;111(2):230-7.17214458 10.1021/jp066041k
The short and intermediate time scale dynamics of the photoacid pyranine (1-hydroxy-3,6,8-pyrenetrisulfonic acid, commonly referred to as HPTS) are studied with visible pump-probe spectroscopy in various solvents to elucidate the nature of its proton-transfer kinetics in water. The observed time dependences of HPTS are compared with those of the methoxy derivative, MPTS. A global fitting procedure is employed to model both the spectral shift (Stokes shift) caused by solvent reorganization and deprotonation of pyranine in water. Three distinct time-dependent features can be clearly identified. They are the Stokes shift (1 ps in H(2)O and 1.5 ps in D(2)O), followed by the deprotonation processes, which gives rise to a biexponential decay of the protonated species with time constants (in H(2)O) of 3 and 88 ps. By the use of a model previously discussed in the literature, the biexponential process can be interpreted as an initial deprotonation step followed by the longer time scale process which separates the resulting ion pair. The results presented here are consistent with some of the previous reports but unambiguously identify and quantitatively measure the Stokes shift as a separate and distinct phenomenon from the deprotonation process, in contrast to other reports that have suggested that all short time (a few picoseconds) dynamics are merely a Stokes shift.
A slowing down of proton motion from HPTS to water adsorbed on the MCM-41 surface
Phys Chem Chem Phys 2016 Jan 28;18(4):2658-71.26705542 10.1039/c5cp04548g
We report on the steady-state and femtosecond-nanosecond (fs-ns) behaviour of 8-hydroxypyrene-1,3,6-trisulfonate (Pyranine, HPTS) and its interaction with mesoporous silica based materials (MCM-41) in both solid-state and dichloromethane (DCM) suspensions in the absence and presence of water. In the absence of water, HPTS forms aggregates which are characterized by a broad emission spectrum and multiexponential behavior (τsolid-state/DCM = 120 ps, 600 ps, 2.2 ns). Upon interaction with MCM41, the aggregate population is found to be lower, leading to the formation of adsorbed monomers. In the presence of water (1%), HPTS with and without MCM41 materials in DCM suspensions undergoes an excited-state intermolecular proton-transfer (ESPT) reaction in the protonated form (ROH*) producing a deprotonated species (RO(-)*). The long-time emission decays of the ROH* in different systems in the presence of water are multiexponential, and are analysed using the diffusion-assisted geminate recombination model. The obtained proton-transfer and recombination rate constants for HPTS and HPTS/MCM41 complexes in DCM suspensions in the presence of water are kPT = 13 ns(-1), krec = 7.5 Å ns(-1), and kPT = 5.4 ns(-1), krec = 2.2 Å ns(-1), respectively, The slowing down of both processes in the latter case is explained in terms of specific interactions of the dye and of the water molecules with the silica surface. The ultrafast dynamics (fs-regime) of the HPTS/MCM41 complexes in DCM suspensions, without and with water, shows two components which are assigned to intramolecular vibrational-energy relaxation (IVR) (∿20 fs vs. ∿.8 ps), and vibrational relaxation/cooling (VC), and charge transfer (CT) processes (∿ ps without water and ∿ ps with water) of the adsorbed ROH*. Our results provide new knowledge on the interactions and the proton-transfer reaction dynamics of HPTS adsorbed on mesoporous materials.
Effects of Cations on HPTS Fluorescence and Quantification of Free Gadolinium Ions in Solution; Assessment of Intracellular Release of Gd3+ from Gd-Based MRI Contrast Agents
Molecules 2022 Apr 12;27(8):2490.35458689 PMC9032885
8-Hydroxypyrene-1,3,6-trisulfonate (HPTS) is a small, hydrophilic fluorescent molecule. Since the pKa of the hydroxyl group is close to neutrality and quickly responds to pH changes, it is widely used as a pH-reporter in cell biology for measurements of intracellular pH. HPTS fluorescence (both excitation and emission spectra) at variable pH was measured in pure water in the presence of NaCl solution or in the presence of different buffers (PBS or hepes in the presence or not of NaCl) and in a solution containing BSA. pKa values have been obtained from the sigmoidal curves. Herein, we investigated the effect of mono-, di-, and trivalent cations (Na+, Ca2+, La3+, Gd3+) on fluorescence changes and proposed its use for the quantification of trivalent cations (e.g., gadolinium ions) present in solution as acqua-ions. Starting from the linear regression, the LoD value of 6.32 µM for the Gd3+ detection was calculated. The effects on the emission were also analyzed in the presence of a combination of Gd3+ at two different concentrations and the previously indicated mono and di-valent ions. The study demonstrated the feasibility of a qualitative method to investigate the intracellular Gd3+ release upon the administration of Gd-based contrast agents in murine macrophages.