Month: December 2019

Synergy between HPTLC and HPLC-DAD for the investigation of wine-making by-products

By Tatiana Bernardi,* Olga Bortolini, Alessandro Massi, Gianni Sacchetti, Massimo Tacchini and Carmela De Risi*

This post is a summary of recently published results concerning the investigation of wine-making by-products by synergistic use of high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography with diode array detection (HPLC-DAD) [1].
This work fits into a more general context related to the revalorization of agri-food by-products, which is extremely current for both environmental and economic reasons [2,3].
With particular regard to wine-making by-products, it is known that they are sources of biologically relevant compounds, mostly polyphenols, such as flavonoids and phenolic acids [4,5]. Thanks to the biological activities of these constituents (e.g. antioxidant, antiinflammatory, antimicrobial, antibacterial) [6-9], grape wastes could be re-used for applications in various fields, such as the nutraceutical and pharmaceutical ones. Accordingly, it is desirable to develop methods for isolation, characterization and quantification of molecules contained in winery by-products, especially in view of their re-utilization on a large scale.
Phenolic compounds present in natural matrices have been investigated by HPTLC [10-14], sometimes in conjuction with HPLC [15,16]. To the best of our knowledge, each method has been generally used to confirm the results of the other, and no further information was inferred by comparing the data obtained from each technique.
Based on these literature findings and our experience on the analysis of complex samples through HPTLC [17-20], we have developed the work in question, proving that HPTLC and HPLC-DAD are really complementary in both determining and quantifying flavonoid and non-flavonoid compounds within wine-making by-products.
Going into more detail, we investigated two types of winery by-products, that is pomace (stalks, seeds, skins) and desiccated seeds deriving from both Italian red (Lambrusco) and white (Trebbiano) cultivars of Vitis vinifera L. After being dried (ventilated oven, 70 °C, 24 h), these materials were subjected to solid-liquid extraction procedures (Naviglio® extractor [21], ultrasounds) using pure ethanol-water mixtures with increasing water content up to 100% [9].
Next, HPTLC fingerprint evaluation [22] served to establish the best extraction solvent (50/50 pure ethanol-water) and identify the major classes of organic compounds (anthocyanins, phenolic acids, non-anthocyanic flavonoids) present in the extracts on the basis of analyte spots intensity and color, after derivatization with NP/PEG reagent (Figure 1A).
Concurrent investigation based on HPTLC/mass spectrometry (MS) interface and band comparison method gave indications on the type of molecules. In particular, only anthocyanins could be detected by HPTLC/MS: cyanidin-3-O-glucoside (kuromanin), delphinidin-3-O-glucoside (myrtillin) and malvidin-3-O-glucoside (oenin) were the main species (Figure 1B), with the latter being found in greater quantity. It is worthwhile noting that chromatogram zones corresponding to phenolic acids and non-anthocyanic flavonoids gave no noticeable signals in the mass spectra, probably due to their low concentration in the extracts, given that more than 250 ng/zone are required to have a significant visualization with MS interfaced to HPTLC [23].
On the other hand, phenolic acid (i.e. caftaric acid) and non-anthocyanic flavonoid (i.e. quercetin 3-O-glucuronide, quercetin-3-O-glucoside) compounds were assigned by comparison of their bands (color and retardation factor) with those of available standards (Figure 1C).

Figure 1. A: typical image of HPTLC separations for optimization of extraction solvent and identification of compound classes. B: chemical structures of the main anthocyanins identified by HPTLC-MS. C: HPTLC identification of non-anthocyanic flavonoids and phenolic acids by band comparison method (1: quercetin 3-O-rutinoside, 2: quercetin 3-O-glucoside, 3: quercetin 3-O-rhamnoside, 4: quercetin 3-O-glucuronide, 5: quercetin 3-O-galactoside, 6: extract from red grape pomace, 7: extract from white grape pomace, 8: extract from red grape seeds, 9: caftaric acid, 10: kaempferol, 11: quercetin).

Subsequent HPLC-DAD screening of red grape pomace extracts led to identify just anthocyanins at 520 nm (Figure 2A) and surprisingly, the same molecules have been identified in the extracts deriving from seeds incorporated in red grape pomace samples. This was in contrast with literature reports indicating that anthocyanins are not found in grape seeds [24], but it may be assumed that the contact between seeds and skins during grape processing may lead to this consequence.
Importantly, the absence in the chromatograms of peaks related to phenolic acids and non-anthocyanic flavonoids (wavelenght range 250-370 nm) has proven not to depend on sample concentration, indeed both types of compounds could not be detected while increasing the concentration of the injected sample solutions up to the limits of filterability (70 mg/mL).
In light of the above, what obtained by HPLC-DAD seemed to contradict the information given by HPTLC fingerprint, however some possible explanations could be offered. Possibly, the concentration of either phenolic acids or non-anthocyanic flavonoids, even when present, is so low that they may be lost during sample pre-treatment (filtration) before HPLC-DAD analysis. On this point, it has to be noted that the supposed low concentration of these species matches the results observed in HPTLC-MS studies. Moreover, it cannot be excluded that the limit of detection (LOD) of DAD was responsible for the incomplete (false) analytical response.
Suitably optimized HPLC-DAD chromatographic conditions (modified Flamini and Favretto [25] method) allowed to identify delphinidin-3-O-glucoside (myrtillin), cyanidin-3-O-glucoside (kuromanin) and malvidin-3-O-glucoside (oenin) in the extracts, confirming what we found by way of HPTLC-MS. Then, quantification via calibration curve method (Figure 2B) showed that oenin was the most abundant species, followed by myrtillin and kuromanin in this order (Figure 2C). At the same time, it was possible to quantify the content of unidentified anthocyanins building on the calibration curve of oenin (Figure 2D).

Figure 2. A: HPLC-DAD chromatogram of a typical red grape pomace extract at 520 nm. B: calibration curves of kuromanin, myrtillin and oenin. C: quantification of kuromanin, myrtillin and oenin; D: quantification of unidentified anthocyanins.

In addition, HPLC-DAD chromatograms provided an indication of the presence of proanthocyanidins (baseline drift at 280 nm), but their identification was not possible.
Instead, HPTLC helped with that, in fact inspection of representative extracts deriving from red (white) grape pomace and seed materials showed that catechin was the only compound present, as demonstrated by densitogram profiles (Figure 3A). Additional HPTLC analysis corroborated this observation, with higher molecular weight derivatives (i.e. epigallocatechin, epigallocatechin gallate) being not found at all. Finally, catechin content was shown to be about the same in both red and white grape pomace extracts (Figure 3B).

Figure 3. A: (a) densitograms of four representative extracts (tracks 1-4), epicatechin standard solution (track 5) and catechin standard solution (track 6); (b) detail of standard catechin densitogram; (c) detail of standard epicatechin densitogram; (d) illustrative densitogram of sample extracts showing the complete lack of epicatechin. B: calibration curve of catechin for red and white grape pomace extracts (top), and quantification of catechin in red and white grape pomace extracts (bottom).

To conclude, this work confirmed the potential of both HPTLC and HPLC-DAD for the analysis of complex mixtures, also showing that the advantages of one technique could be exploited to compensate for the limits of the other. Thus, typical features of HPTLC (minimal sample manipulation, i.e. dilution, low LOD, multiple derivatization) have helped achieve a greater extracts characterization (anthocyanins, non-anthocyanic flavonoids, phenolic acids) that supplemented the limited information given by HPLC-DAD (anthocyanins) for same sample types. Although HPLC-DAD gives the advantage of monitoring matrices content in typical wavelength ranges with just one injection, either sample pre-treatment (filtration) or LOD of detector might provide incomplete (false) results, in apparent contradiction with HPTLC response. This being the case, each technique is not just for confirming the data provided by the other, as usually done, but one adds to the other.

Acknowledgments: Thanks are due to the regional funding POR-FESR 2014–2020 within the project “Valorizzazione sostenibile degli scarti della filiera vitivinicola per l’industria chimica e salutistica (VALSOVIT)”.
For more information about this study, go to the DOI: 10.3390/molecules24193416



Tatiana Bernardi (right) graduated in Chemistry at the University of Ferrara in 1997. She worked at the Basell laboratories as Quality Controller of polymers production (1998-1999), then she moved back to the University of Ferrara and was hired as a technician. She received a master's degree in “Sugar and Alcohol Technology” from the University of Ferrara (2003) and her PhD in “Applied Bio-Catalysis and Industrial Microbiology” from the University of Bologna (2008). She is presently technician (level C5) at the Department of Chemical and Pharmaceutical Sciences of the University of Ferrara. Her main research interests are in the analytical field, with particular regard to high-performance thin-layer chromatography (HPTLC, AMD-OPLC) and high-performance liquid chromatography (HPLC), in mass spectrometry applications, and also include industrial microbiology and fermentation developments and monitoring.

Scopus Author ID: 8343635300

Carmela De Risi (left) graduated in Chemistry at the University of Ferrara (1992) and received her PhD in Organic Chemistry in 1996. That same year, she joined the group of Prof. P. Vogel at the University of Lausanne (Switzerland) where she spent one year research as a grant holder. Then, she moved back to the University of Ferrara where she performed postdoctoral studies (1997-1999). Since November 1999 she has been Research Associate at the University of Ferrara. Her main research interests focus on synthesis and modification of biologically active compounds, general synthetic methodologies, organocatalysis, biomass valorization.

Scopus Author ID: 6603965163


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Posted by in Chemistry

Fertility and Wine

By Shaun R. McCann

The making of wine is an organic process and has similarities with pregnancy. Many characteristics of a wine are determined during the growing period. Mineral deficiencies can inhibit full development of the grape. Fungal infection or hailstones can destroy a crop shortly before harvest, just as intrauterine infection can be devastating during a pregnancy. Viral infections can affect grapes and the growing foetus. When to harvest/deliver requires careful consideration. As the time to harvest approaches oenologists (and gynaecologists) become agitated. A lot of things can happen in the remaining few days. Decisions made in these few days can have deleterious or beneficial effects on the outcome. Oenologists will do two things: measure the sugar content of the grapes and taste the grape juice. The oenologist has many scientific aids available to help in making the decision to harvest, but there is no substitute for experience and knowledge of the soil and previous behaviour of that clone of grapes. Similarly, the obstetrician may have many tools, such as electronic foetal heart monitoring, but there is no test or machine that can replace experience. Harvesting too soon or too late will result in a suboptimum wine. Delivering a baby too early or too late may result in prematurity or foetal distress. The oenologist and the obstetrician may not be fully aware of the final results for many years after all the work is done.

Since the documentation of the foetal alcohol syndrome (FAS) by Jones and Smith in 1973 women who consume a very moderate amount of wine e.g. a glass of wine with their evening meal, have become pariahs. There is no doubt that the FAS exists and causes devastating toxicities to infants including irreversible brain damage and growth retardation. However, the deleterious effects of alcohol on the developing foetus seem to be maximal in the first trimester and diminish thereafter. So, it is probably best for pregnant women to desist from alcohol during the first trimester of pregnancy but the evidence that moderate wine consumption (1–2 glasses of wine per week) during the second and third trimester are harmful is difficult to elicit. I have asked a number of distinguished Professors of Obstetrics and Gynaecology to provide me with the evidence that moderate wine consumption by pregnant women, who are otherwise healthy and well fed is harmful to their health or that of the foetus, but so far, I have received no satisfactory reply. Meta-analyses conclude that the evidence that moderate wine consumption is harmful to foetal development is sparse. What about the effects of wine consumption on fertility? Published results are confusing. One study found that a moderate intake of wine shortened the time to pregnancy in couples not using any form of contraception. As they say, ‘Whether this is an effect of wine itself or the characteristics of the wine drinker is not known’. Perhaps another explanation is that after a glass of wine they felt in the mood!

Another study claimed that it is not possible to demonstrate any deleterious effect on socioemotional difficulties or cognitive deficits on 5-year-old children whose mothers had engaged in light wine drinking during pregnancy. So, it looks like light wine consumption during pregnancy has no measurable deleterious effects on the baby. However, as Antonio said in William Shakespeare’s The Merchant of Venice: ‘The devil can quote scripture for his purpose’. Although the data are conflicting, probably the best advice to give pregnant women is to abstain from alcohol during the first trimester and if she must, then limit wine consumption to a couple of glasses per week during the second and third trimester. Some authorities e.g. The CDC (Center for Disease Control) in Atlanta, Georgia, in the United States, and some national bodies recommend total abstinence from alcohol during pregnancy based on little or no evidence. Luckily, it is not a decision I will ever have to make!
I presume we all worry that we will end up with some form of dementia if we live long enough. There may be some good news. Ruth Peters and colleagues from Imperial in London suggest that moderate alcohol consumption in early adult life might offer some protection from dementia. Sigfried Weyerer and colleagues, in the same journal in an article entitled, ‘Current alcohol consumption and its relationship to incident dementia: results from a 3-year follow-up study among primary care attenders aged 75 years and older’, concluded that light-to-moderate alcohol consumption is inversely related to incident dementia. Let’s hope it’s true and even if it isn’t, a glass of wine or two daily helps to make the world bearable especially in these turbulent times.
One may well be skeptical about the claims made for the purported health benefits of wine, but beware of questioning Greeks bearing gifts, and surely nobody can, in good faith, deny the mood enhancement that moderate wine drinking can bring? True, when it is abused it can have disastrous and life-destroying consequences, especially when in conjunction with addictive tendencies. However, when consumed in sensible quantities, it can be and is, as the Greeks realized, one of life’s great pleasures.

Shaun R. McCann
MB, FRCPI, FRCPEdin, FRCPath. Hon.FTCD. Hon Life Member EBMT. Hon Life Member EHA
Recipient of EHA Education and Mentoring Award June 2019.
Professor Emeritus of Haematology and Academic Medicine, St James’s Hospital and Trinity College, Dublin.


DOB 1-10-46
Graduate of University College Dublin, 1970, MB, B. Ch, B.A.O.
Specialist Medical Fellow University of Minnesota, U.S.A.1974-76
Lecturer/Senior Lecturer in Haematology Trinity College Dublin, 1976 -1984
Consultant Haematologist and Head of Department St James’ Hospital/TCD 1984-2009.
Performed the first successful allogeneic bone marrow transplant for leukaemia in Ireland in 1984.
Developed a molecular haematology research laboratory in SJH.
A test to measure chimaerism using PCR of STRs was developed by Dr Mark Lawler, Professor Pete Humphries and myself and was widely used internationally by Stem Cell Transplant Teams.
Introduced an art intervention (Open Window) into the stem cell transplant unit and this study, of which I was the PI, was published in 2011. The study played a major role in the choice of QoL as the ‘theme’ for the European Hematology Association (EHA) in 2012-2013.
Appointed George Gabriel Stokes Professor of Haematology @ Trinity College Dublin, 1995-2010.
Vice-President of IACRLRLD (International Association for Comparative Research on Leukemia and Related diseases).1994-1997.
Appointed National Medical Director, Blood Transfusion Service (by invitation of the Minister for Health, Michael Noonan TD) 1995-1996 during the Hepatitis C outbreak.
First President of the Haematology Association of Ireland.
Author of over 200 articles in peer-reviewed medical journals, numerous book chapters and two medical textbooks.
Clinical Cases Uncovered (CCU), Haematology, Wiley/Blackwell. ISBN 978-1-4051-8322-2
This is currently being developed as an interactive platform by EHA.
Appointed Professor of Academic Medicine, School of Medicine, Trinity College Dublin 2006- 2011
On the Editorial Board of Bone Marrow Transplantation and Hematology and reviewer for The British Journal of Haematology, Blood, The Journal of Blood and Bone Marrow Transplantation, Cancer, Bone Marrow Transplantation.
Author of ‘invited editorials’ on Wine and Haemopoietic Cell Transplant (HCT) for the journal Bone Marrow Transplantation since July 2018.

1970. O’ Ferrall Gold Medal in Surgery, St Vincent’s Hospital, Dublin.
1976-77 Registrar’s prize RAMI (Royal Academy of Medicine in Ireland), Section of Pathology.
1986 St Luke’s Lecture, RAMI.
2011 Guest speaker Biological Society, TCD.
2016 Guest lecturer Gemelli Hospital, Rome, Italy.

An Immodest Proposal-Wine and Health (under the pseudonym Giovanni Morelli) ISBN 978-1-78280-207-5.
A history of Haematology: from Herodotus to HIV. Oxford University Press. ISBN 978-0-19-871760-7
Blood Matters. European Hematology Association June 2019. ISBN 978-90-823759-1-6

Selected puplications

Molecular Haematology

M. Lawler, S.R.McCann, E Conneally and P Humphries. Chimaerism following allogeneic bone marrow transplantation: detection of residual host cells using the polymerase chain reaction. Br J Haematol 1989, 73: 201-210.
M. Lawler, P Humphries and S.R.McCann. Evaluation of mixed chimaerism by in vitro amplification of dinucleotide repeat sequences using the polymerase chain reaction. Blood 1991, 77: 2504-2514.
P.V. Browne, M Lawler, J O’Riordan, P Humphries and S.R. McCann. Early detection of leukaemic relapse after bone marrow transplantation.
Bone Marrow Transplantation 1991, 7: 167-169.
P.V. Browne, M Lawler , P and S.R.McCann . Donor-cell leukemia after bone marrow transplantation for severe aplastic anemia.
N Engl J Med. 1991;325:710–713.
S.R. McCann, M Lawler and A Bacigalupo. Recurrence of Philadelphia chromosome-positive leukemia in donor cells after marrow transplantation for chronic granulocytic leukemia. Leuk Lymphoma. 1993; 10: 419-425.
S.R McCann and M. Lawler. Mixed Chimaerism; detection and significance following BMT. Bone Marrow Transplantation. 1993, 11: 91-94.
S McCann and E Wright. Donor cell leukemia: perhaps a commoner occurrence than we thought. Bone Marrow Transplantation. 2003; 32:455-457.
Hertenstein B, Hambach L, Bacigalupo A, Schmitz N, McCann S, Slavin S, Gratwohl A, Ferrant A, Elmaagacli A, Schwertfeger R, Locasciulli A, Zander A, Bornhäuser M, Niederwieser D, Ruutu T; Chronic Leukaemia Working Party of the European Group for Blood and Marrow Transplantation. Development of leukemia in donor cells after allogeneic stem cell transplantation--a survey of the European Group for Blood and Marrow Transplantation (EBMT). Haematologica. 2005. 90 (7): 669-675.

N Gardiner, M Lawler, J O’Riordan, M De Arce, P Humphries and S.R. McCann. Persistent donor chimaerism is consistent with disease-free survival following BMT for Chronic Myeloid Leukaemia. Bone Marrow Transplantation. 1997, 20; 3: 235-241.
O’Riordan J; GardinerN; OMeara A; Molloy K; Lawler M; Stallings R; McCann S.R 
Immunotherapy for Relapsed
 Leukaemia. British Journal of Cancer, 1998, 78: 127-127
N Gardiner, M Lawler J O’Riorda, J.M De Arce and S.R. McCann. Monitoring lineage specific chimaerism allows early prediction of relapse following donor lymphocyte infusions for relapsed chronic myeloid leukaemia. Bone Marrow Transplantation.1998, 21: 711-716
N Gardiner, S.R. McCann, J O’Riordan, M Lawler. Chimerism following Donor Lymphocyte Infusion for Chronic Myeloid Leukemia. Blood. 1999, 93: 2748-2749
S.R. McCann, K Gately, E Conneally and M lawler. Molecular response to Imatinib Mesylate following relapse after allogeneic SCT for CML. Blood 2003, 101; 3: 1200-1200.
D O’Shea, G Crotty, P Carroll, E conneally, S McCann and M.J. Neat. Clonal karyotypic abnormalities in philadelphia negative cells of CML patients treated with Imatinib: is it underreported and does it have any clinical significance? Br J haematol. 2004. 123 (3): 367-369.
P.J. Hayden, F Keogh, M ni Chongaile, M Carroll, M Crowley, N Fitzsimon, N Gardiner, E Vandenberghe, J O’Riordan and S.R. McCann. A single-centre assessment of long-term quality of life status after sibling allogeneic stem cell transplantation for chronic myelid leukaemia in chronic phase. Bone Marrow Transplantation. 2004;34: 545-556.
S.R. McCann. Chronic Myeloid Leukaemia: a paradigm for malignancy or just a strange disease. Sultan Qaboos Univ Med J 2012, 12; (4): 422-428.

Aplastic Anaemia
A Locascuilli, L van.t;Veer, A Bacigalupo, J hows, M.T. van Lint, E Gluckman, C Nissen, S McCann, I Vossen, H Schrezenmeier, W Hinterberger, and A Martin. Treatment with marrow transplantation or immunosuppression of childhood acquired severe aplastic anaemia. Bone Marrow Transplantation 1990, 6; 3: 211-217.
J Marsh, G Socie, H Schezenmeier, A Raghavachar, A Tichelli, Pljungman, S.R. McCann, P Marin, J.M. Hows, A Bacigalupo. EBMT working Party for Severe Aplastic Anemia. Haemopoietic growth factors in Aplastic Anaemia: a cautionary note. Lancet 1994, 344; 8916: 172-173.
S.R. McCann, A Bacigalupo, E Gluckman, W Hinterberger, J Hows, P Ljungman, P Marin. Graft rejection and second marrow transplants for acquired Aplastic Anaemia.EBMT WP for SAA. Bone Marrow Transplantation 1994, 13: 233-237.
Aplastic Anemia: Pathophysiology and Treatment. Eds H Schrezenmeier and A Bacigalupo. HLA-identical sibling bone marrow transplantation to treat Severe Aplastic Anemia. S.R.McCann, J Passweg, R Storb, and J Deeg. Cambridge University Press. 1999.
A Tichelli, G Socie, J Marsh, R Barge, N Frickhofen, S McCann, A Bacigalupo, J Hows, P Marin, D Nachbaur, A Symdonidis, J Passweg and H Schrizenmeier, EBMT working party for Severe aplastic Anemia. Outcome of pregnancy and disease course among women with Aplastic Anemia treated with Immunosuppression. Ann Int Med 2002; 137: 164-172.
C. Marsh, S.E. Ball, P Darbyshire, E.C.Gordon-Smith, A.J. Kiedan, A Martin, S.R. McCann, J Mercieca, D Oscier, A.W, Roques, J.A. Yin. Guidelines for th diagnosis and treatment of acquired aplastic anaemia. Br J Haematol 2003, 123; (5): 782-801.
D O'Donghaile, P. J. Hayden, S. L. McCarron, E. M. Doyle, M. Lawler, P. V. Browne, E. Conneally, E. Vandenberghe, S. R. McCann. Marrow aplasia developing 13 years after HLA-identical sibling allogeneic transplantation for chronic myeloid leukaemia: successful treatment with antithymocyte globulin and peripheral blood stem cell infusion from the original donor. European Journal of Haematology. 2006, 76; 3: 258-260.
Lawler M, McCann SR, March JC, Ljungman P, Hows J, Vandenberghe E, O'Riordan J, Locasciulli A, Socie G, Kelly A, Schrezenmeier H, Marin P, Tichelli A, Passweg JR, Dickenson A, Ryan J, Bacigalupo A; Severe Aplastic Anaemia Working Party of the European Blood and Marrow Transplant Group. Serial chimerism analyses indicate that mixed haemopoietic chimerism influences the probability of graft rejection and disease recurrence following allogeneic stem cell transplantation (SCT) for severe aplastic anaemia (SAA): indication for routine assessment of chimerism post SCT for SAA. J Haematol. 2009; 144(6):933-4
A Piccin, S McCann, G Socie, R Oneto, A Bacigalupo, A Locasciulli, J March, H Survival of patients with documented autologous recovery after SCT for severe aplastic anaemia: a study by the WPSAA of the EBMT. Bone Marrow Transplantation. 2010; 45 (6): 1008-1013.
Quality of Life in treatment of Cancer/Haematological Malignancies.
McCabe C, Begley C, Collier S, McCann S. Methodological issues related to assessing and measuring quality of life in patients with cancer: Implications for Patient Care. European Journal of Cancer Care, 2007.17(1): 56-62.
Denis Roche, Philip Napier, Brian Maguire and Shaun McCann. A clinically useful artwork? Between a Dialogical and a Relational Approach to art in a clinical environment. The International Journal of the Arts in Society. 2008. vol 3
F Hegarty, C McCabe, D Roche and S McCann. Using multimedia technology to help combat the negative effects of protective isolation on patients: The Open Window Project-an engineering challenge. Journal of Visual Communication in Medicine. 2009,32; 3:72-77.
McCabe C, Roche D, Hegarty F, McCann S. Open Window’: a randomized trial of the effect of new media art using a virtual window on quality of life in patients’ experiencing stem cell transplantation Psycho-Oncology. 2011, doi. 1002/pon.2093.
S.R. McCann. Can an art intervention influence the quality of life in hospitalized patients? Haematologica, 2013, 98 (1): 4-6.
C Chomienne, M Guenova, A Hagenbeek, C Lacombe, S McCann, S Salek, J Geissler, I van der Beek. Quality of Life in Haematology: EHA theme of the year…and years to come. Haematologica 2013, 98 (1): 2-3.
Shaun R. McCann. Can we influence the ‘quality of life’ for patients in hospital? Health Environments Research and Design Journal. 10 (1): 170-171. 2016.

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