ULTRAVIOLET RADIATION-INDUCED PHOSPHOLIPASE A2 ACTIVATION OCCURS IN MAMMALIAN CELL MEMBRANE PREPARATIONS

Ultraviolet erythema in human skin is mediated in part by membrane derivatives of arachidonic acid (AA). UVA (320–400nm) and UVB (290–320nm) have been shown to induce release of AA from intact mammalian cells in culture. In order to investigate the mechanism of this release we examined the effect of UVA and UVB on release of [³H] AA from membrane preparations of murine fibroblasts. C3H 10T1/2 cells were prelabelled for 24 h with [³H] AA. The membrane fractions of the cells were separated after lysis by differential centrifugation. The membranes were irradiated in suspension and the [³H] AA released from the membranes was determined by scintillation spectroscopy of supernatants3–4 h after irradiation. Both UVA and UVB induced release of AA from the membrane preparations. The response to UVB was small but significant, reaching levels approximately 150% of control release at doses of 1,200-4,000 J/m². The response to UVA was larger; doses of 2.5-5.0 J/cm² induced release equal to twice control (200%) levels, while doses of10–20 J/cm² induced maximal release at levels approximately 400% of control. Time course studies with UVB and UVA showed maximal release at 4 h after irradiation. When the membrane preparations were incubated with a polyclonal anti-phospholipase A₂ antibody the UV induced release of [³H] AA was completely inhibited in both UVB (1200 J/m²) and UVA (10 J/cm²) treated cells. These data suggest that activation of phospholipase A₂ is responsible for the UV induced release of AA in mammalian cells and that the mechanism of this activation is due, in part at least, to direct photon-membrane interaction.     

Ultraviolet radiation stimulates the release of arachidonic acid from mammalian cells in culture

Abstract— C3H 10T½ cells in culture were prelabelled with [³H]arachidonic acid and exposed to UVB radiation. Almost immediately after irradiation cells released labelled arachidonate metabolites into media in a dose dependent manner. This release was inhibited by removing calcium ions from the system and by the addition of dexamethasone and parabromophenacyl bromide to the system. This suggests that the UVB stimulated release of arachidonic acid from membrane phospholipids is, in part, mediated by a phospholipase A₂ enzyme system. Thin layer chromatographic examination of media extracts revealed a dose dependent UVB stimulation of prostaglandin production by cultured cells.     

COMPARISON OF EFFECTS OF UVA, UVB AND UVC ON GROWTH AND VIABILITY OF MITOGEN-STIMULATED HUMAN PERIPHERAL BLOOD CELLS

Abstract— Peripheral blood mononuclear cells were irradiated with UVA, UVB or UVC. The highest exposure dose used in each waveband reduced the number of viable cells to one-third the control cell population after 3 days in culture. Exposure of these cells to half as much UV from each waveband resulted in an equivalent or greater degree of inhibition of their proliferative response to mitogen as measured by lymphoblast transformation, [³H]-thymidine uptake and viable cell number on day 3 in culture. The pattern of inhibition was distinct for each waveband. UVA interfered with blastogenesis on the first 2 days of culture at doses which had considerably less effect on viable cell number. UVA also depressed the first round of DNA synthesis, which was detectable on the second day of culture. By day 3 in culture, however, the UVA-induced reduction in both the number of lymphoblasts and the uptake of [³H]-thymidine was a direct reflection of reduced numbers of viable cells. UVB did not interfere with blastogenesis in mitogen-stimulated cultures to the same degree as did UVA. Only the highest dose of UVB depressed blast transformation more than viable cell number on day 1; by day 2 lower doses were also inhibitory. In contrast UVC had little effect on blastogenesis at any time; a reduced number of lymphoblasts observed on days 2 and 3 in culture was a direct reflection of a reduced number of viable cells rather than a reduced percent of these cells undergoing blast transformation. As with UVA-irradiated, mitogen-stimulated cells, [³H]-thymidine uptake was also depressed in both UVB and UVC irradiated, mitogen-stimulated cells on day 2. However, only UVB continued to depress DNA synthesis more than viable cell number after 3 days of culture. These results suggest that UVA, UVB and UVC may interfere with any one or more of the signals involved in the response to mitogen, be they the recognition of mitogen by T cells or accessory cells, the transformation of lymphocytes into lymphoblasts or the activation of lymphoblasts to synthesize DNA.     

THE EFFECT OF THE ANTIPSORIATIC DRUG METABOLITE ETRETIN (Ro 10-1670) ON UVB IRRADIATION INDUCED CHANGES IN THE METABOLISM OF ARACHIDONIC ACID IN HUMAN KERATINOCYTES IN CULTURE

[¹⁴C]Arachidonic acid was avidly incorporated into human keratinocytes in culture and following exposure to UVB irradiation of 9 mJ/cm² (erythemally effective, EE) substantial amounts of ¹⁴C-radiolabel were released from the cells. The release of radiolabel was accompanied by a decrease in the labelling of phosphatidylethanolamine whereas the labelling of triacylglycerols and cholesteryl esters was increased. Keratinocytes produced significant amounts of prostaglandin E₂ (PGE₂) and following UVB irradiation of 9 mJ/cm² (EE) the formation of prostaglandin E₂ was increased.
Etretin (Ro 10-1670), the active metabolite of the antipsoriatic drug etretinate (Ro 10-9359), affected significantly neither the total release of radiolabel induced by UVB nor the formation of prostaglandin E₂. However, in the presence of etretin the UVB irradiation induced transfer of [^l4C]arachidonic acid into triacylglycerols and cholesteryl esters was not increased as much as in the corresponding experiments without etretin. On the basis of the present study it appears that etretin does not interfere with the release of arachidonic acid in amounts which could be related to the therapeutic effects of the combination of retinoids with UVB irradiation (Re-UVB) in the treatment of psoriasis.     

Fast-atom bombardment tandem mass spectrometry of [13C]arachidonic acid labeled phospholipid molecular species

A method employing stable isotope labeling and fast-atom bombardment (FAB) tandem mass spectrometry has been developed to directly assess events of biosynthesis and metabolism of arachidonic acid containing phospholipid molecular species by cells carried in culture. Mast cells, cultured with [¹³C]linoleic acid, converted this precursor into arachidonic acid which was then incorporated into cellular phospholipids. Over a 24 hour period, the extent of label enrichment in each arachidonate-containing phospholipid molecular species was monitored by using negative FAB ionization with selected reaction monitoring. Specific incorporation of [¹³C₁₇] labeled arachidonate was determined from the ratio of the carboxylate anions at m/z 320 and 303, which correspond to [¹³C₁₇]arachidonate and unlabeled arachidonate, respectively, produced by collision-induced dissociation of each specific molecular anion. The use of [¹³C]linoleic acid as a precursor of arachidonic acid avoids the problem of changing the endogenous pool size by directly adding labeled arachidonic acid. Measurement of the [¹³C₁₇]label also avoids interferences from endogenous isobaric fatty acids that are naturally present at low levels.     

UVA irradiation induces energy-independent phospholipid-flip in mammalian plasma membrane

Translocation from the outer to the inner membrane leaflet (flip) of phospholipids after ultraviolet A (UVA) irradiation was investigated in Chinese hamster ovary cells. Fluorescent 1-palmitoyl-2-[6-[(7-nitro-2-1,3-benzox- adiazol-4-yl)amino]caproyl]-sn-glycero-3-phosphoserine (NBD-labeled phosphatidylserine [NBD-PS]) was used to assay transbilayer lipid movement. A marked increase in flip of NBD-PS was observed immediately after low-dose UVA irradiation which was not lethal and returned to the basal level after 6 h. UVA-induced flip was not attributed to the increase of permeability by UVA irradiation because cells that were negative for staining with propidium iodide also showed increased flip of NBD-PS. Furthermore, the enhancement was independent of adenosine 5'-triphosphate, demonstrating the lack of involvement of phospholipid translocase. Marked increases were also observed in flip of both NBD-phosphatidylethanolamine and NBD-phosphatidylcholine immediately after UVA irradiation, showing that the increase was independent on the head groups of phospholipids. These findings indicated that UVA changes the flip-flop of phospholipids and that the cell membrane is a molecular and cellular target of UVA.     

脂肪酸离子排斥分配色谱分离中的吸附作用

酸在氢型阳离子交换树脂柱上的分离机理已有研究,Glod等根据离子排斥色谱中的Donnan膜平衡,推导出分配系数K_d与酸离解常数K_a的关系,能大致预测酸的流出顺序,但不能说明为什么pK_a极为相近的有机酸却有较大的保留时间差别.Bafna等从分子吸附的角度考察了某些有机酸在阳离子交换树脂上的吸附平衡和洗脱行为.本文从有机酸离子排     

A methodology for radiolabeling of the endocannabinoid 2-arachidonoylglycerol (2-AG)

The metabolic intermediate and endocannabinoid signaling lipid 2-arachidonoylglycerol (2-AG) has not been readily labeled, primarily because of its instability toward rearrangement. We now detail a synthetic method that easily gives tritiated 2-AG from [5,6,8,9,11,12,14,15-(3)H(N)]arachidonic acid in two steps. We utilized a short chain 1,3-diacylglycerol and proceeded through the "structured lipid" [5',6',8',9',11',12',14',15'-(3)H(N)]2-arachidonoyl-1,3-dibutyrylglycerol, a triacylglycerol that was conveniently deprotected in ethanol with acrylic beads containing Candida antarctica lipase B to give [5',6',8',9',11',12',14',15'-(3)H(N)]2-arachidonoylglycerol ([(3)H]2-AG). The flash chromatographic separation necessary to isolate the labeled 2-acylglycerol [(3)H]2-AG resulted in only 4% of the rearrangement byproducts that have been a particular problem with previous methodologies. This reliable "kit" method to prepare the radiolabeled endocannabinoid as needed gave tritiated 2-arachidonoylglycerol [(3)H]2-AG with a specific activity of 200 Ci/mmol for enzyme assays, metabolic studies, and tissue imaging. It has been run on unlabeled materials on over 10 mg scales and should be generally applicable to other 2-acylglycerols.     

A novel ligand-free palladium catalytic system with SO3H-functional ionic liquids as cocatalyst for amidocarbonylation reaction

Effects of Lewis acid BF3·OEt2, and BrCnsted acids TsOH, CF3COOH, H3PO4, and HCIO4 as cocatalyst respectively on the ligand-free palladium-catalyzed amidocarbonylation were investigated. SO3H-functional ionic liquids 1-methyl-3-（4-sulfonic acid）butylimidazolium hydrosulfate [MIm（CH2）4803H][HSO4] and 1-methyl-3-（4-sulfonic acid）butylimidazolium triflate [MIm（CH2）4SO3H][OTf] were firstly employed as cocatalysts instead of these Lewis acid and Brφnsted acids. By using a ligand-free and weak corrosive catalyst in situ prepared form PdBr2, LiBr.H2O, and [MIm（CH2）4SO3H][OTf], the arnidocarbo- nylation of benzaldehyde, acetamide, and CO could proceed smoothly and afford N-acetyl-α-phenylglycine with yield of 58% in [C6mim]PF6 medium.     

Biochemical characteristics, metabolism, and antitumor activity of several acetylated hexosamines

We have synthesized several potential inhibitors and/or modifiers of the carbohydrate portion of plasma membrane glycoconjugates. These include fluorinated and actylated analogs of D-glucosamine, D-galactosamine, and D-mannosamine. These compounds have been tested to determine their effects on both[14C] glucosamine and [3H] leucine incorporation into glycoconjugate and on cell growth and viability using P-288 murine lymphoma cells maintained in tissue culture. The most cytotoxic agent tested was 2-acetamido-2-deoxy-1,3,4,6-tetra-O-acetyl-beta-D-glucopyranose or simply beta-pentaacetylglucosamine which prevented cell growth at 10(-4)-10(-3) M. beta-Pentaacetylglucosamine cytotoxicity was correlated with its high lipid solubility, having an octanol/water partition coefficient of 0.424 as compared with 0.278 for the alpha-anomer and 0.017 for N-acetylglucosamine. In vitro metabolism studies with [4C]- and/or [3H]-labeled pentaacetylglucosamine have indicated intracellular de-O-acetylation leading to the biosynthesis of UDP-N-acetylglucosamine, followed by the incorporation of this sugar into cellular glycoprotein. Concomitant with the formation of increased amounts of this nucleotide sugar, intracellular UTP and CTP pools fell to one third normal within 3 h after the administration of 1 mM pentaacetylglucosamine. At present it is unclear whether the cytotoxicity of beta-pentaacetylglucosamine or other similar agents is due to alterations in nucleotide and nucleotide-sugar pools causing a decrease in energy charge and polynucleotide biosynthesis or is due to a direct effect on membrane glycoconjugate biosynthesis.     

Effect of an inhibitor of squalene epoxidase, NB-598, on lipid metabolism in Hep G2 cells.

NB-598, a potent inhibitor of squalene epoxidase, inhibited cholesterol synthesis from [14C]acetate and induced intracellular squalene accumulation in Hep G2 cells. NB-598 inhibited cholesterol synthesis from [14C]acetate, [3H]mevalonate, and [3H]squalene, but not from [3H]2,3-oxidosqualene in Hep G2 cells. It reduced cholesterol ester synthesis remarkably in the absence of exogenous cholesterol. This compound did not have any effect on the synthesis of ubiquinone and dolichol. When Hep G2 cells were prelabeled with micellar [3H]cholesterol, NB-598 did not affect the excretion of bile acid incorporated from [3H]cholesterol. However, NB-598 decreased the secretion of free and esterified cholesterol, triacylglycerol, and phospholipids, and increased the secretion of squalene. NB-598 is thought not only to inhibit cholesterol synthesis, but also to inhibit the secretion of lipids.     

Effect of growth phase on the Escherichia coli response to ultraviolet-A radiation: influence of conditioned media, hydrogen peroxide and acetate

The results reported herein indicate that the ultraviolet-A (UVA) radiation-induced effects in Escherichia coli depend on its growth phase. Stationary-phase cells recover faster from a sub-lethal UVA exposure and have a higher resistance to lethal effect of the radiation than exponential growing cells. Although pre-incubation in spent medium supernatant increased the resistance of log-phase cells to lethal UVA effects, this pre-treatment considerably prolonged the duration of the radioinduced sub-lethal growth delay. The aim of the present study was to investigate the effect exerted by the E. coli conditioned media and evaluate the influence of nutritional stress, hydrogen peroxide and acetate. Pre-incubated in conditioned medium, cells in exponential growth phase were irradiated and the induced effects were compared with those found when catalase, high culture densities and acetate were employed. Unexpectedly, the duration of the growth delay in cells submitted to these treatments was shortened in comparison with control cells incubated in conditioned medium with no modifications. Lengthening of the growth delay was mimicked when exponentially growing cells were incubated in fresh medium supplied with 5 microM H(2)O(2). The effects of spent medium on wild type and rpoS mutant strains were similar, indicating that this response is independent of RpoS controlled functions. We assumed that an oxidative component of the spent medium, probably H(2)O(2), could be involved in the observed phenomenon. This effect is specific of E. coli and independent of rpoS.     

A radiometric assay method for aromatase activity using [1 beta-3H]16 alpha-hydroxyandrostenedione.

[1 beta-3H]16 alpha-Hydroxyandrostenedione (16 alpha-OHA) (715 mCi/mmol) was prepared from commercially available [1 beta-3H]androstenedione (A) by the microbiological method with Streptomyces roseochromogenes and its structure and purity were determined by chromatographic and reverse isotope dilution methods. When [1 beta-3H]16 alpha-OHA was incubated with human placental microsomes and reduced nicotinamide adenine dinucleotide phosphate (NADPH), 3H2O-release into the medium was dependent upon protein concentration and incubation time. An apparent Km and Vmax of the microsomal aromatase for the [1 beta-3H]substrate were 650 nM and 34 pmol/min/mg protein, respectively. In this assay, aromatase activity could be determined as low as 0.1 nmol estrogen formation/min/mg protein. 3-Deoxyandrostenedione, a potent competitive inhibitor of the A aromatization, also blocked the 16 alpha-OHA aromatization in a competitive manner with Ki of 15 nM.     

Metabolic fates of L-tryptophan in Saccharomyces uvarum (Saccharomyces carlsbergensis).

The metabolism of L-tryptophan by Saccharomyces uvarum (carlsbergensis) was investigated by simultaneous measuring of fluxes through kynureninase, through transaminases and into protein using L-[methylene-14C] and L-[side chain-2,3-3H]tryptophan. In yeasts cultivated in synthetic medium (S medium), the flux into protein was predominant, closely followed by the flux leading to 2-3H liberation. The proportion of L-tryptophan metabolized via the latter flux increased over 10-fold (75% of total tryptophan metabolized) as the concentration of L-tryptophan was raised from 5 x 10(-5) to 5 x 10(-4) M. L-Tryptophan metabolized via the kynureninase flux was less than 5% of total tryptophan metabolized. In yeast extract-polypepton-glucose medium (YPG medium), more tryptophan was incorporated into protein than in the S medium. Contribution of the kynureninase flux remained very low. Tryptophan metabolism via each flux changed depending on the growth phase. 2-3H liberation was shown to be primarily due to tryptophol synthesis by high performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR), indole-3-acetic acid and kynurenic acid also contributing to 2-3H liberation but to a much lesser extent. 2-3H liberation increased dose-dependently at tryptophan concentration higher than 10(-5)M, while the kynureninase flux reached its plateau at 10(-5)M. Formation of tryptophol and indole-3-acetic acid via indole-3-pyruvic acid and indole-3-acetaldehyde with indole aldehyde as a by-product was confirmed using exogenous tryptophan metabolites with indole rings.     

Determination of leukotrienes and prostaglandins in [14C] arachidonic acid labelled human lung tissue by high-performance liquid chromatography and radioimmunoassay

A liquid chromatographic method for the determination of 14C-labelled prostaglandins, leukotrienes and other lipoxygenase products formed by human lung tissue is described. In this paper we report our problems identifying these substances when 3H- or 14C-labelled compounds are compared with measurements of the mass by absorption or radioimmunoassay. Furthermore, some preliminary results of [14C] arachidonic acid labelled human lung tissue, stimulated by the Ca-ionophore A23187, show that, of the lipoxygenase products, mostly leukotriene B4 like compounds are formed and less leukotriene C4, E4 and D4. Relatively large amounts of hydroxyeicosatetraenoic acids are present. The main cyclooxygenase products are thromboxane B2, 6-ketoprostaglandin F1 alpha and prostaglandin D2.     

Preparation of 7-hydroxy-2-oxoindolin-3-ylacetic acid and its [13C2], [5-n-3H], and [5-n-3H]-7-O-glucosyl analogues for use in the study of indol-3-ylacetic acid catabolism

An improved synthesis of 7-hydroxy-2-oxoindolin-3-ylacetic acid via the base-induced condensation reaction between oxalate esters and 7-benzyloxyindolin-2-one is described. 7-Benzyloxyindolin-2-one was prepared in four steps and 50% overall yield from 3-hydroxy-2-nitrotoluene. The yield of the title compound from 7-benzyloxyindolin-2-one was 56%. This route was used to prepare 7-hydroxy-2-oxoindolin-3-yl[13C2]acetic acid in 30% yield from [13C2]oxalic acid dihydrate. The method could not be extended to the preparation of the corresponding [14C2]-compound. However, an enzyme preparation from Zea mays roots catalysed the conversion of carrier-free [5-n-3H]indol-3-ylacetic acid with a specific activity of 16.7 Ci mmol-1 to a mixture of 7-hydroxy-2-oxo[5-n-3H]indolin-3-ylacetic acid and its [5-n-3H]-7-O-glucoside in ca. 3 and 40% radiochemical yield respectively. The glucoside was converted into the 7-hydroxy compound in 80% yield by means of beta-glucosidase.     

Protective effects of cyanidin-3-O-beta-glucopyranoside against UVA-induced oxidative stress in human keratinocytes

Ultraviolet-A (UVA) radiation causes significant oxidative stress because it leads to the generation of reactive oxygen species (ROS), leading to extensive cellular damage and eventual cell death either by apoptosis or necrosis. We evaluated the protective effects of cyanidin-3-O-beta-glucopyranoside (C-3-G) against UVA-induced apoptosis and DNA fragmentation in a human keratinocyte cell line (HaCaT). Treatment of HaCaT cells with C-3-G before UVA irradiation inhibited the formation of apoptotic cells (61%) and DNA fragmentation (54%). We also investigated antioxidant properties of C-3-G in HaCaT cells against ROS formation at apoptotic doses of UVA; C-3-G inhibited hydrogen peroxide (H2O2) release (an indicator of cellular ROS formation) after UVA irradiation. Further confirmation of the potential of C-3-G to counteract UVA-induced ROS formation comes from our demonstration of its ability to enhance the resistance of HaCaT cells to the apoptotic effects of both H2O2 and the superoxide anion (O2*-), two ROS involved in UVA-oxidative stress. Furthermore, in terms of Trolox Equivalent Antioxidant Activity, C-3-G treatment led to a greater increase in antioxidant activity in the membrane-enriched fraction than in the cytosol (55% vs 19%). The protective effects against UVA-induced ROS formation can be attributed to the higher membrane levels of C-3-G incorporation. These encouraging in vitro results support further research into C-3-G (and other anthocyanins) as novel agents for skin photoprotection.     

Inhibition of apical membrane enzyme activities and protein synthesis by gentamicin in a kidney epithelial cell line LLC-PK1.

The mechanism of a gentamicin-induced decrease in apical membrane enzyme activities was investigated in LLC-PK1 cells. Increasing activities of apical membrane enzymes (alkaline phosphatase, aminopeptidase, and gamma-glutamyltransferase) were markedly suppressed by gentamicin during growth in culture. On the other hand, a lesser effect was observed when the activities of these enzymes were decreasing or relatively constant. Gentamicin treatment decreased the maximal enzyme activities of alkaline phosphatase and aminopeptidase, indicating that the number of active enzyme molecules in the apical membrane was decreased by gentamicin. [3H]Leucine incorporation in LLC-PK1 cells was inhibited by gentamicin in a dose-dependent manner, followed by a reduction of total protein. In addition, a well-known protein synthesis inhibitor, cycloheximide, also decreased the apical enzyme activities. These results suggest that the inhibition of protein synthesis by gentamicin is a possible cause of the decreased activities of apical membrane enzymes in LLC-PK1 cells. The inhibition of protein synthesis may be related to the nephrotoxicity induced by aminoglycoside antibiotics.     

Utility of isolated hepatocytes and radio-HPLC-MSn for the analysis of the metabolic fate of 19-nortestosterone laurate in cattle.

The metabolic fate of 19-nortestosterone laurate in cattle was investigated to evaluate target analyte(s) appropriate to surveillance for illicit use as a growth promoting agent. Bovine hepatocytes were incubated with either [3H]19-nortestosterone laurate (19-NTL; 4-estren-17 beta-laurate-3-one) or [3H]19-nortestosterone (19-NT; 4-estren-17 beta-ol-3-one; nandrolone). Hepatocyte medium was extracted with solid phase C18 media and analysed by narrow bore radio-HPLC-MSn (LCQ, Finnigan) to evaluate the structure of metabolites of 19-NTL and 19-NT. Radio-HPLC of hepatocyte medium extracts following incubation with [3H]19-NTL confirmed that the first step of biotransformation in liver was hydrolysis of the fatty acid ester to release [3H]19-NT, which, in turn, was converted into a range of metabolites of diverse polarity. Hydrolysis of hepatocyte medium extracts with beta-glucuronidase (Helix pomatia) indicated that some of these metabolites were glucuronide or sulfate conjugates. Structural analysis of unconjugated metabolities by positive-ion atmospheric pressure chemical ionisation MS2 and comparison with available reference preparations indicated biotransformation of 19-NT to 4-estren-17 alpha-ol-3-one, 4-estren-3, 17-dione (major metabolite after 1 h), n-hydroxy-4-estren-3, 17-dione, n-hydroxy-4-estren-17-ol-3-one, 5 beta-estran-3 alpha-ol-17-one (noretiocholanolone) and 5 beta-estran-3 alpha, 17 beta-ol (major metabolite after 4 h). Conjugated metabolites were analysed by electrospray ionization, which revealed the presence of glucuronide conjugates of alpha-(trace) and beta-epimers of 19-NT, n-hydroxy-4-estren-3, 17-dione, n-hydroxy-4-estren-17-ol-3-one and 5 beta-estran-3 alpha, 17 beta-diol. These studies provide a clear indication of the route of hepatic metabolism in the bovine, which may now be readily substantiated by reference to samples, such as urine or bile, derived from animals treated with unlabelled 19-NTL.     

软配位原子效应在锌和镉分离中的应用

研究了锌和镉在二（１,１,３,３－四甲基丁基）膦酸（ＨＭＢＰ）及二（１,１,３,３－四甲基丁基）巯基膦酸（ＨＭＴＰ）／环己烷－硝酸体系中的萃取行为。以软配位原子－硫取代ＨＭＢＰ分子中的羟基氧原子而得到的ＨＭＴＰ对镉具有良好的选择性,以ＨＭＴＰ为萃取剂,仅一次萃取可以实现锌和镉的定量分离。